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Food

Anthony F. Gross,* Peter S. Given, Jr., and Albert K. Athnasios

Nabisco Brands, Inc., Corporate Technology, Box 1943, East Hanover, New Jersey 07936-1943

This review covers the literature for approximately the Sulfite was oxidized to sulfate with sulfite oxidase producing
period from October 1984 to October 1986. This is the first peroxide which was then reduced by NADH in the presence
review by the present authors and we continue to use the basic of NADH peroxidase by Beutler (6A) with subsequent spec-
format of the authors of the last review (23P), as well as the trophotometric measurement of NADH consumption. Re-
practice of citing domestic and more widely circulated foreign sidual sulfite in shrimp was determined by Cooper et al. (12A)
journals over the less accessible ones. This is a critical review using ion chromatography; result was 97% of AOAC Moni-
and not an attempt to provide an all-inclusive bibliography. er-Williams method finding. Damiani and Burini (14A) de-
The chemical abstract reference is now included in most of tected nitrite in milk by reaction with 2,3-diaminonaphthalene
the literature cited. to form naphthotriazole which was measured fluorometrically.
DeVries et al. (16A) combined selective distillation with se-
ADDITIVES lective redox titration to measure sulfite. Eek and Ferrer (18A)
Review articles pertaining to additives consist of the fol- determined nitrate and nitrite in water and ham using ion
lowing: Arneth and Witzgall (3A) compiled thin-layer chro- exchange high-performance liquid chromatography (HPLC)
matography (TLC) methods for the analysis of all commonly with ultraviolet (UV) detection, as did Eggers (19A) for cured
used meat additivies; the chemistry and analysis of sulfite and meats. Grekas & Calokerinos (23A) described a continuous
sulfur dioxide in foods was reviewed by Barnett (4A); and flow apparatus for the detection of sulfite and sulfur dioxide
Modderman (38.4) reviewed the chemistry of sulfite, its re- in beverages and air samples using molecular emission cavity
action with food components and problems encountered analysis. Holak (27Al) developed quantitative methods to
during measurement. extract and differentiate free and combined sulfite and

212R 0003-2700/87/0359-212R$06.50/0 © 1987 American Chemical Society

 FOOD

Anthony F. Gross, Director Methods Devel- flowing stream by gas diffusion, decolorizes malachite green.
opment. Corporate Technology, Nabisco Simultaneous determination of benzoate, sorbate, and four
Brands, Inc. (B.S. 1951, College of the City
of New York), supervises the methods de- p-hydroxybenzoic acid esters in meat using reverse-phase
HPLC was achieved by Ali (1 A). Similar work was reported
velopment and problem solving efforts of
the Scientific Services Department. His by Garcia-Regueiro et al. (2JA). Eichler and Rubach (20A)
work experience includes analysis of food- measured sorbate, benzoate, p-hydroxybenzoic acid esters,
stuffs. pesticide residue analysis, instrumen- formate, and propionate in a single step using isotachophoresis.
tal analysis, and the development of analyti- Matsunagi et al. (36A) reported the simultaneous determi-
cal methods. He has served as an associ- nation of sorbate, benzoate, p-hydroxybenzoic acid esters and
ate referee for the Association of Official saccharine by ion-pair HPLC using hexadecyltrimethylamine.
Analytical Chemists. Presently he serves as
an industry advisor to the U.S. Delegation of
Ion-pair extraction with tri-n-octylamine was used by
the Codex Alimentarius Committee of Analy- Puttemans et al. (40A) with reverse-phase HPLC to measure
sis and Sampling. He is a member of the sorbate, benzoate, and saccharin in yogurt. Haddad and
Association of Official Analytical Chemists and a member of the American Jackson (24A) reported simultaneous determination of as-
Chemical Society. corbate, bromate, and metabisulfite in bread improver using
ion exchange HPLC with both UV and conductivity detection.
Peter S. Given, Jr., Ph.D., is Group Leader A similar technique was applied by Yamamoto et al. (54A)
in the Fundamental Science Department of to measure potassium bromate in bread. Rao et al. (41A)
Nabisco Brands. Inc. (B.S. 1975, Syracuse determined qunine in soft drinks by spectrophotometry of the
University, Ph.D. 1981, State University of complex formed between quinine and alizarin brilliant violet
New York). He is responsible for fundamen- R at 578 nm, Reijenga et al. (42A) measured quinine using
tal research programs on food ingredients isotachophoresis. Quinine, hydroquinine, saccharin, and so-
which include interactions between macro- dium benzoate concentrations in soft drinks were measured
molecules and low molecular weight com-
pounds and the physical-chemical behavior
by Valenti (53A) using direct injection into reverse-phase
of lipids. Current focus involves high-resolu- HPLC with UV detection at 254 nm. Chikamoto and Maitani
tion NMR applications. Prior activities wilh (10A) determined ethanol and propionic acid levels in baked
the company include development of products simultaneously by steam distillation followed by GC
analytical methods with emphasis on HPLC, analysis. Yanai (56A) developed a test paper for ethanol by
investigative work on consumer inquiries, impregnating paper with Monascus red dye which reveals color
and chemometric approaches to fingerprint in the presence of ethanol vapor.
analysis and discrimination.
Argoudelis (2A) developed a cation exchange HPLC method
for the simultaneous determination of aspartame, caffeine,
Albert K. Athnaslos. Senior Research Asso- saccharin, and benzoate in soft drinks. Ammonium phosphate
ciate, Corporate Technology, Nabisco mobile phase and UV detection at 214 nm were utilized.
Brands, Inc., East Hanover, NJ [B.Sc. Aspartame and its degradation products were determined by
(1956), Alexandria University, Egypt; M.S. using gradient elution reverse phase HPLC by Cross and
(1962), North Carolina State University, Ra- Cunico (13A). A /i-cyclodextrin bonded phase HPLC column
leigh, NC], manages the GC/MS and NMR was used by Issaq et al. (30A) to measure aspartame in soft
groups of the Analytical Methods Develop- drinks. Aspartame was also measured by TLC and ninhydrin
ment Department and directs problem-solv-
visualization by Sherma et al. (46A), Limacher and Tanner
ing efforts in nonroutine and critical analy-
ses. His interests include analytical methods (34A) measured cyclamate using GC separation of the reaction
development and troubleshooting and the
adduct of cyclamate with 20% sulfosalicyylic acid followed
separation and elucidation of structure of by treatment with hydrochloric acid and hydrogen peroxide.
chemical constituents of food and natural Yamauchi et al. (55A) constructed an amperometric enzyme
products by applying chemical, chromato- (L-glutamate oxidase) electrode to measure L-glutamic acid.
graphic, and spectrometric techniques. He
has authored several publications on the analysis of mycotoxins, carbohy-
Indyk and Woollard (29A) determined the antioxidants,
drates. and lipids in foods, on natural flavor constituents in tobacco, and on
BHA, BHT, TBHQ, and tocopherols, in tallow and vegetable
the isolation and structural determination of medicinally active chemical con-
oil by normal-phase HPLC with UV detection at 280 nm.
stituents from natural products and developed methodologies for intermediate BHA, BHT, and TBHQ in edible oils were measured by
process control and for the analysis of a wide spectrum of food chemical Kitada et al. (31 A) using reverse-phase HPLC with ampero-
components including intense sweeteners, emulsifiers, antioxidants, and trace metric detection. A specific spectrophotometric method for
food contaminants. Organizational activities include ACS, AO AC, and AOCS BHA was developed by Komaitis and Kapel (32A) by reacting
where he is a member of ihe Mycotoxins and Uniform Methods Committees. BHA with AvV-dimethyl-p-phenylenediamine and measuring
at 550 nm. Stoyke et al. (49A) developed an extraction
technique and a GC method for ETDQ, the active component
in the feed antioxidant XAX-M.
measured extract concentrations by using differential pulse A qualitative test for the presence of galactose-containing
polarography. The selectivity of malachite green for detecting thickeners in meat was described by Bauer and Stachelberger
the presence of sulfite in poultry was reevaluated by Mallinson (5A) where the meat is acid hydrolyzed and the liberated
et al. (35A); some interferences were found. Extracted nitrate galactose measured enzymatically. Brumley et al. (8A) dis-
was utilized for quantitative nitration of benzene with sub- cussed the limits and resolution of OH negative ion chemical
sequent GC-thermal energy analysis by Ross and Hotchkiss ionization mass spectrometry (MS) analysis of polysorbates
(43A). Schwedt and Baeurle (44A) compared photometry with (polyoxyethylated fatty acid esters of sorbitan). Resolution
p-rosaniline, HPLC, and enzymatic analyses of sulfite (as of fatty acid esters of sucrose on open tubular columns with
sulfur dioxide) in foods; the results varied widely according supercritical carbon dioxide mobile phase was described by
to food matrices. Schwedt (45A) describes the use of com- Chester et al. (9A). Mitsuhashi et al. (37A) developed an
mercial test strips for simultaneous measurement of nitrite, enzymatic regimen to measure propylene glycol in a variety
nitrate, sulfur dioxide, ascorbic acid, and potassium. Color of foods using glycerol dehydrogenase and glycerol kinase. A
intensity was measured by reflectometry. Silva et al. (47A) GC method for simultaneous quantitative determination of
developed a method for sequential analysis of nitrate and methylated fatty acids and silylated sterols from orange juice
nitrite in a continuous flow/extraction system by formation was reported by Stack et al. (48A).
of an organic copper complex with nitrate and atomic ab- Simultaneous determination of monensin, narasin, and
sorption spectrophotometric analysis of the complexed copper. salinomycin in feeds using reverse-phase HPLC and postco-
Nitrite either was oxidized to nitrate with Ce(IV) and mea- lumn reaction with vanillin was described by Blanchflower
sured by copper complexation or was reduced to nitrogen with et al. (7A). DeVries et al. (15A) developed an automated
sulfamic acid and passed undetected. Sulfur dioxide was headspace GC-ECD (electrochemical detection) method for
distilled into formaldehyde and then measured by use of ion methyl bromide in wheat, flour, cocoa, and peanuts. The
exchange HPLC with conductivity detection by Sullivan and fungicides biphenyl, 2-hydroxybiphenyl, and thiabendazole
Smith (50A). Sullivan et al. (5JA) detected sulfite by flow were detected by Gieger (22A) in fresh fruits using reverse-
injection analysis where sulfur dioxide, isolated from the phase HPLC with fluorescence detection. Heikes (25A) im-

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 213 R
FOOD

proved a 1-h purge and Tenax-TA trap method for ethylene Galensa et al. (84B) used HPLC for detecting grape juice
dibromide determination by GC-ECD. A microbiological addition to orange juice by separating and determining nar-
method for systemic fungicide residue determination was ingin hydrolysis products. Adulteration in commercial cran-
adapted by Kovacs (33A) to incorporate several bacterium and berry juice drinks and concentrates was detected by Hong et
yeast species to measure a variety of fungicides. Duncan et al. (118B) by HPLC analyses for nonvolatile acids and an-
al. (17A) compared HPLC-electrochemical detection with thocyanidine profiles. UV-visible spectral measurements were
GC-MS-single ion monitoring (SIM) for the assay of salso- used to measure pigment concentration, polymeric color, and
linol, dopamine, 3,4-dihydroxyphenylacetic acid, 3,4-di- percent polymeric color. Adulterated samples were detected
hydroxyphenylethanol, and norepinephrine in foods and by having nonvolatile organic acid profile indicative of added
beverages, and found GC-MS-SIM far superior. Hobson- malic and/or citric acid. Substantial levels of delphinidin and
Frohock (26A) performed tissue residue studies of medicinal malvidin, neither of which are present in cranberries in sig-
additives, amprolium, arprinocid, clopidol, dimetridazole, and nificant amounts, were indicative that grape skin extract was
sulfaquinoxaline, in broiler chickens and turkeys using GC the added colorant. Kauschus et al. (136B) found that analysis
and HPLC. The determination of tetracyclines and macrolide of the sugar components of water-soluble polysaccharides in
antibiotics in various meats was carried out by Hoshino et al. orange juice by CGC showed the arabinose/galactose ratio to
(28A) using a cyano column for HPLC analysis. Newsome be remarkably constant. This value (0.50-0.53) offers an
(39A) developed an enzyme-linked immunoassay for triad- additional criterion for evaluating the authentic composition
imeton in fruits and vegetables and found good agreement of orange juice. Siewek (235B) reported that adulteration of
with results from an established GC procedure. Cook et al. black currant juice by red currant juice may be detected by
(11 A) determined packaging headspace oxygen, nitrogen, and the fact that the red juice contains a flavonol glycoside that
carbon dioxide distributions by developing a special sampling is absent in the black juice. HPLC, NMR, and spectrometric
device, collection of gases in an evacuated sampling valve, and methods were used to characterize the structure of the com-
sample injection into a GC fitted with a thermal conductivity pound. Siewek et al. (236B) detected the adulteration of grape
detector. Carbonation levels in beverages were measured by juice and of alcoholic beverages from grape juice with fig juice
Szerenyi and Ringleib (52A) by passing the beverage over one by determining of flavone-C-glycosides which do not occur
side of a gas-permeable membrane and then measuring the in grapes using HPLC. An addition of 5% fig juice can be
carbon dioxide content in a diluent gas passed over the op- detected with certainty. Wald et al. (283B) detected adul-
posite side of the membrane continuously using either thermal teration of black currant products with blackberry juice by
conductivity or infrared. determination of flavonoids by HPLC. Wallrauch (284B)
wrote a review with 10 references on the use of amino acid
ADULTERATION, CONTAMINATION, content of fruit juices to detect adulteration by other juices,
DECOMPOSITION to detect dilution with water or other fluids, and to identify
Gilbert (90B) published a book on The Analysis of Food fruit varieties used for juice manufacturing. The variability
Contaminants, and wrote a review (91B) on the advantages of proline content in a number of juices was tabulated.
and disadvantages of mass spectrometry in identification of Wrolstad (295B) analyzed apple juice samples from several
food components. Folkes (79B) wrote a review on the de- countries for sugar and nonvolatile acid content and discussed
termination of contaminants in food and the impact of de- the use of the results to detect adulteration of the juice. Stable
C isotopes were also determined and recommendations were
velopments in instrumental analysis on methodology and
limits of determinations. Specific examples of contaminants made for improving the analytical methods and broading the
discussed are As, Pb, Hg, Cd, organochlorine pesticides, analysis data in the detection of juice adulteration.
polynuclear aromatic hydrocarbons, polychlorinated biphenyls, Zyren et al. (307B) reported on the interlaboratory varia-
vinyl chloride monomer, styrene, and nitrosamines. bility of methods used for detection of economic adulteration
Immunoassay techniques for measuring veterinary drug of apple juice. The most important test for determining
residues in farm animals, meat, and meat products were re- adulteration was the total malic acid/L-malic acid ratio.
viewed by Heitzman (115B). Horman (120B) published a Fumaric acid, a minor contaminant in synthetically produced
review on mass spectrometry and food safety and the use of malic acid, shows promise as an indictor of economic adul-
MS to determine natural toxiants and contaminants in foods. teration. Volonterio et al. (281B) reported a sensitive method
Reid published two reviews (210B, 211B) on the extraction for determining nonbiogenic acetic acid in fermentation
and cleanup of contaminants and toxicants from food for mass vinegar by radiochemical 14C counting.
spectrometric analysis. Contribution of Mass Spectrometry Frangipane et al. (82B) detected by GC isoglucose-adul-
to Food Safety was the subject of a review by Stan (244B). terated honey by determining the isomaltose/maltose ratio.
Shepherd (288B) reviewed size exclusion and gel chroma- White et al. (293B) reported that the detection of beet sugar
tography and applications to the cleanup of food samples for adulteration of honey by quantitation of oligosaccharide-bound
contaminant analysis. galactose by galactose oxidase treatment of the higher sugar
Adulteration of natural lime oils was detected by Alessandro fraction is useful to screen honeys with normal stable C isotope
et al. (3B) by glass capillary GC-MS. Brause et al. (27B) ratio values. However, additional testing is required for
detected adulteration in apple juice and orange juice by a confirmation of adulteration.
chemical matrix method combining isotope analysis, sugar Adulteration of durum wheat pasta with nondurum wheat
profiles, UV absorbance, glucose/fructose ratio, total sug- gliaden proteins was detected by Burgoon et al. (34B) using
ars/Brix ratio, mineral, and trace elements, and determination aluminum lactate polyacrylamide gel electrophoresis at pH
of trace organic compounds of beet sugar and invert sugar by 3.1. Pastas adulterated with as little as 5% hard red winter
GC/MS was used to detect adulteration of orange juice es- wheat flour could be detected with this method.
pecially with water and pulp wash. Adulteration of apple juice Cow milk adulteration of buffalo milk and buffalo mozza-
can be determined by sugar profile, 13C ratio, chlorogenic acid, rella cheese was detected by the mobility of casein by poly-
malic acid, proline, and sorbitol content. Bricout et al. (29B) acrylamide gel electrophoresis as reported by Beghi et al.
used the isotopic ratios 13C/12C ad 2H/1H for detecting the (14B). Farag et al. (77B) used GC for the qualitative and
addition of cane and beet sugars and high fructose com syrup quantitative determination of fatty acids of authentic buffalo
to orange juice in an effort to control its authenticity. milk, cow milk, and buffalo milk adulterated with cow milk,
Cohen et al. (46B) collected data on the composition of and by application of a simple regression equation for par-
Israeli orange juice, grapefruit juice, orange peel, and grapefruit ticular acids, adulteration of buffalo milk with 5% cow milk
peel. Significant differences were found between the juice and could be detected. Krause et al. (143B) reported that the
peel in the content of isocitric acid, total flavonoids, chlorides, presence of cow milk in sheep, goat, or buffalo milk could be
phosphates, chloramine T, and arginine. Significance of these detected at 1-2% by measuring the amount of 7-2-casein in
findings in detecting juice adulteration with peel extract solids the mixture by isoelectric focusing. The presence of cow milk
was discussed. Doner (66B) reported on carbon isotope ratios could also be detected in sheep, goat, or buffalo cheeses. The
in natural and synthetic citric acid as indicators of lemon juice presence of cow milk whey in whey cheeses was detected by
adulteration. Fang et al. (76B) reported the use of HPLC and determining d-lactoglobulins by isoelectric focusing. In ricotta
ion chromatography for the analysis of citrus fruit juice and cheese, where the whey proteins are heat denatured, addition
passion fruit juice of Taiwan for free amino acids. Fruit juice of a strong reducing agent was necessary to resolubilize the
authenticity was inspected by amino acid distribution patterns. proteins before isoelectric focusing. Pollman (203B) used
214 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
 FOOD

calcium, phosphorus, magnesium, and lactose indexes to detect tected and estimated the presence of linseed oil in rape oil.
adulteration in grated cheese. Mariani et al. (157B) developed a procedure to detect
Carnegie et al. (38B) used a new HPLC method to monitor adulteration of oils and margarines by determination of their
the adulteration of cooked beef products with meat from other mono- and diglyceride content. The glyceride classes were
species. The ratio of the histidine dipeptides anserine and separated by TLC on silica gel with hexane-ether-formic acid
carnosine, which are present in skelectal muscle, are so dif- (100:100:3), and silylated glycerides were further fractionated
ferent between sheep, cattle, horse, and kangaroo that de- by GC on a 5% SE 52 glass capillary column with temperature
tection of adulteration can be rapidly achieved by HPLC on programming.
Partisil SCS columns with 0.2 M lithium formate at pH 2.9 Sheppard et al. (232B) applied various analytical methods
as mobile phase. To obtain a definitive identification of the for the detection, identification, and quantitation of vegetable
adulterant, it was necessary to also examine the electrophoretic oil adulteration in ice cream by determination of total fat
mobility of myoglobin in SDS gels. Imre et al. (125B) reported content, sterols, short- and long-chain fatty acids, vitamin E,
that polyacrylamide gel disk electrophoresis of beef, horse, Reichert-Meissl values, and Polenske values. All methods,
and donkey muscle proteins gave species-specific patterns that except total fat determination were capable of detecting
can be used for identification. The method can be used to vegetable oil adulteration. Sterol determination was the most
detect 5% adulteration of ground beef with horse or donkey effective and versatile measurement because it provided in-
meat. Kumudavally et al. (145B) employed TLC and GC to formation not only on the detection and extent of adulteration
analyze free fatty acids from canned meat curries prepared but also on the possible identity of the adulterant. Chadha
from fresh and spoiled mutton and admixtures of the two. et al. (39B) used capillary GC-FID for detection and iden-
Analysis of the free fatty acid fraction revealed a 10-12-fold tification of brominated vegetable oils in soft drinks. Gubman
increase in palmitic, stearic, and oleic acids. Zitko (306B) et al. (10IB) determined phosphatides in plant oils colori-
published a review with 98 references on organic contaminants metrically.
in fish and shellfish and methods for their determination. Gibbrellic acid residues on fruits was determined by Cruces
Firestone et al. (78B) reported on the detection of adul- et al. (51B) by synchronous scanning derivative spectrofluo-
terated or misbranded olive oil products that have been rometry. L-Pyroglutamic acid as an indicator of decomposition
shipped to the United States during 1983-1984. Triscornia or abuse of processed foods was determined spectrophoto-
et al. (267B) reported on the possibility of verifying the gen- metrically in molasses, wastewater, and catsup by Ekstoem
uineness of olive oils by determination of the alkanal content et al. (71B). Uranium was determined in commercial milk
of edible virgin and lampante olive oils and solvent extracts samples by Gamboa et al. (85B) by measuring the a particles
of olive marc. Mariani et al. (158B) detected solvent extracted emitted after irradiation, chemical etching, and electrochem-
marc oil in virgin (pressed) olive oil by separating the wax ical etching, and Luan (152B) determined traces of arsenic
constituents with silica gel column chromatography or TLC, in soft drinks by oscillographic polarography.
followed by determining the wax esters by capillary GC. Skatole and indole were determined in the back fat of pigs
Statistical procedures for the discrimination between marc by Garcia-Regueiro et al. (86B) using RP-HPLC. Gardiner
oils and virgin, lampante, or rectified oils were proposed on et al. (87B) used an improved modification of an official AOAC
the basis of the C number (34-46) and amounts of wax esters method for extraction and detection of filth in hard and soft
present. Addition of 10% marc oil to pressed oil was de- cheese. Hatanaka et al. (109B) described a rapid and si-
tectable. Dirinck et al. (65B) reported that total aldehyde or multaneous analysis of hippuric acid and benzoic acid in
hexanal as main contributor to aldehyde content when de- fermented milk or raw milk using RP-HPLC.
termined by dynamic headspace analysis may be used for Microbial growth in stored wheat was assessed by Kaminski
objective measurement of rancidity intensity in oils. et al. (134B) by GC of the formed volatiles like diacetyl,
El-Sayed et al. (72B) detected lard in hydrogenated fat by acetoin, 3-octanone, and 1-octanol. Carbonyls, as a measure
the higher amount of palmitic acid in the 2-position of the of bacterial and fungal infection changes, were measured as
trigycerides, and the saturated/unsaturated acid ratio in the their oximes by photometry, and ergosterol as a measure of
2-position also could be used to distinguish lard from hy- fungus and yeast growth was measured by capillary GC or UV
drogenated oil and shortenings. Gomez et al. (95B) detected spectrometry. Uric acid, free fatty acids, and polar lipids were
mineral oil in vegetable oil by TLC, and analysis of rape oil determined with HPLC by Wetzel et al. (291B) as an objective
denatured with aniline and associated with toxic syndrome criteria for condition of stored wheat.
showed that two oils presented an anamolous hydrocarbon Pentachlorophenol residues in chicken liver and fat were
band with characteristics similar to those of mineral oils. determined by Neidert et al. (180B) by GC-ECD and in ge-
Gopalakrishnan et al. (97B) described a method for detecting latin by Yip (301B) by GC-ECD on a 1% SP-1240 DA column
and estimating adulteration in coconut oil with palmolein by with methane in argon as carrier gas. The method for pen-
treatment of the oil with Carr-Price reagent. The presence tachlorophenol in gelatin was adopted official first action by
of palmolein is indicated by the appearance of a blue-black AOAC.
color, which was determined at 700 nm. A simple method for Phenol was determined in honey by Ogawa et al. (185B)
detecting palmolein adulteration in peanut oil was described using GC-FID. Page (194B) determined acrylonitrile (ACN)
by Motghare et al. (172B). in foods by headspace GC with N/P selective detection. The
Grover (100B) reviewed the presence and detection of toxic method was studied collaboratively for ACN determination
substances, in argemone, castor, karanja, ambadj, and salseed in cold pack cheese, peanut butter, honey butter, and mar-
oils, aflatoxin in peanut oil, and thiocyanate and thiosalidines garine (in plastic containers). The method for (ACN) was
in rape oil. Han et al. (107B) developed a reversed-phase modified by Page et al. (193B) to include packaged luncheon
HPLC method for quantitative determination of adulteration meats.
of sesame oil with soybean oil. The procedure could be ex- Decomposition products of Usal (aspartame HC1), were
tended to detect adulteration of sesame oil with oils other than determined by Prudel et al. (205B) in model systems and
or in addition to soybean oil, e.g., with rape oil. Detection dioxopiperazine determined in soft drinks by HPLC. Usal
of argemone oil adulteration of edible oils was reported by and its decomposition products aspartic acid, phenylalanine
Madhyastha et al. (153B) by extraction of sanquinarine from methyl ester, aspartylphenylalanine, phenylalanylaspartic acid,
an edible oil adulterated at 1% by argemone oil followed by and dioxopiperazine were separated on Separon Si C-18 using
its analysis on an alumina-silica gel minicolumn where it was phosphate buffer (pH 2.1)-methanol (85:15) as mobile phase
detected as a yellow fluorescent band in the silica layer. with UV detection at 200 nm.
Nasirullah et al. (179B) reported a comparative study of three Takarai et al. (250B) determined foreign substances in foods
methods available for the detection of argemone oil in edible and additives (plasticizers) in commercial plastic products by
vegetable oils, a wet method which depends on the formation direct inlet mass spectrometry (DI-MS) with no pretreatment.
of crystalline red-brown chelation compounds of sanquinarine Fibers of cigarette filters were found in commercial shiokara;
and dihydrosanquinarine with ferric chloride could detect ethyl palmitate and ethyl linoleate, and ergosterol were found
argemone at 1 %, and two chromatographic methods (paper in commercial bread.
and TLC) could detect 0.02%. Sauter et al. (215B) determined A method of cleanup by droplet countercurrent chroma-
adulteration of pumpkin seed oil by thin-layer chromatogra- tography (DCC) for food contamination monitoring was de-
phy of the unsaponifiable fraction and examining the pattern veloped by Onji et al. (188B, 189B). Yamazaki et al. (299B)
of unsaponifiable compounds. Manandhar et al. (156B) de- proposed a method for dibutyltin determination in food

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 215 R
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containers and packaging materials. Residual vulcanization HPLC. Aflatoxins in vegetable oils were analyzed by Miller
accelerators (dialkyldithiocarbamates) in baby bottle rubber et al. (166B), the oil samples dissolved in hexane were applied
teats were analyzed by Yamazaki et al. (298B) by GC with to a silica column and washed with ether, toluene, and chlo-
N/P detection. roform; aflatoxins were eluted from the column with chloro-
Vanhaelen-Fastre et al. (279B) qualitatively and quanti- form-methanol (97:3) solvent mixture, and quantitated by
tatively determined the hallucinogenic components of Psilo- TLC and HPLC. Recoveries of aflatoxin Bx standards added
cybe mushrooms by reversed-phase HPLC. to aflatoxin-free oils were between 89.5 and 93.5% with
Mycotoxin analysis was the subject of several review articles. precision of 6.3-8.0%. Orbey et al. (191B) determined afla-
Briantais (28B) published a review with 38 references on toxins Bx, B2, Gx, and G2 in hexane defatted hazelnuts by
analytical methods for determination of mycotoxins, regulatory chloroform extraction, followed by cleanup on a silica gel
aspects, and control of foods. Aflatoxin Bx, aflatoxin Mx, and cartridge and HPLC analysis using coumarin as internal
patulin contaminations were emphasized. Campbell et al. standard. Shepherd et al. (229B) carried out a systematic
(35B) presented a review with 38 references on sampling, investigation of the conditions employed for the postcolumn
sample preparation, and sampling plans of foodstuffs for iodination reaction used to enhance fluorescence sensitivity
mycotoxin analysis. Coker (49B) wrote a review with 138 of aflatoxins Bx and Gx in reversed-phase HPLC. The effects
references on HPLC and other chemical quantification of postcolumn reaction coil tubing dimension, coil tempera-
methods used in the analysis of mycotoxins in foods. Crosby ture, eluent, and reagent flow rates and reagent concentrations
(50B), in a review with 27 references, discussed chemical were examined, and the optimum choice of each was estab-
methods of analysis, for the extraction, cleanup, and deter- lished. The maximum achievable sensitivity was 20 pg of Bx
mination of aflatoxins from animal feeds and foods. The injected on column. Spilmann (240B) studied a modification
advantages and disadvantages of TLC and HPLC were dis- of the AOAC official method for screening aflatoxin in corn
cussed along with newer techniques such as immunoassay. and the feasibility of using it as a quantitative method. Several
Methods for determination of ochratoxin, patulin, and tri- different corn products were analyzed by using the modified
chothecenes were also included as well as problems encoun- method with an average savings of >1 h/sample vs. the CB
tered during collaborative mycotoxin method evaluations. method, with average recoveries of Bx of 94% for the low-level
Park et al. (195B) published a review with 55 references on spiked samples and 108% for the high level. Results obtained
the official methods of analysis of mycotoxins in foods such by analyzing naturally contaminated samples compared fa-
as those approved by the AOAC, IUPAC, AACC, and AOCS. vorably with the CB method. Picogram amounts of aflatoxins
Ueno (274B) published a review with 37 references on de- in milk and cattle were determined by Veres et al. (280B) using
velopment of immunoassay using monoclonal antibodies for radioimmunological analysis. Wei et al. (289B) determined
the detection of mycotoxins in farm products; the principles aflatoxins Bx, B2, Gx, and G2 in infant food by normal-phase
of immunoassay, preparation of antigens and antibodies, and HPLC.
application of the immunoassay in detection of aflatoxins, Whitaker et al. (292B) determined the amount of aflatoxins
trichothecenes, and ochratoxins were discussed. extracted from raw peanuts by using water-slurry modifica-
Candlish et al. (36B) reported on a monoclonal antibody tions of AOAC method 11 for 49 combinations of methanol
to aflatoxin Bx and the detection of the mycotoxin by an concentration and solvent/peanut ratio. The amount of af-
“ELISA” assay which had a sensitivity of 0.2 ppb with a latoxins Bx and B2 extracted from raw peanuts was a function
working range up to 10 ppb for aflatoxin Bx. Ram et al. (206B) of both methanol concentration and solvent/peanut ratio; a
screened naturally contaminated corn and cottonseed samples cubic equation was developed, using regression techniques,
for aflatoxin Bx by a direct competitive ELISA. ELISA es- to describe the combined effects. From the functional relation,
timates for aflatoxin Bx in corn were in good correlation with the predicted methanol concentration and solvent/peanut
values obtained by TLC, and ELISA estimates for aflatoxin ratio were computed to be 60% and 10.8 mL/g of peanuts,
Bx in cottonseed were in good correlation with values obtained respectively. This combination extracted 12.1% more afla-
by HPLC. By use of this ELISA, 36 duplicate sample extracts toxin than did AOAC method II.
can be screened for aflatoxin Bx in less than 2 h. Fremy et al. (83B) established protocols for detecting pi-
The use of ELISA for determining mycotoxins in food was cogram quantities of aflatoxin Mx in milk, yogurt, brick
discussed by Terplan et al. (262B) who used an ELISA assay cheddar, and ripened Brie cheese by ELISA. Good agreement
to determine aflatoxin Bx, aflatoxin Mx, ochratoxin A, and T-2 was found for aflatoxin Mx levels in several naturally con-
toxin in various foods (milk, dried milk, cheese, yogurt, wheat, taminated milk samples analyzed by both ELISA and HPLC.
corn, peanuts, and barley). The results correlated well with Saito et al. (214B) developed a TLC method for determining
those obtained by a TLC method. Ueno (275B) developed aflatoxins Mx and M2 and aflatoxicols I and II in cereals, nuts,
a simple and improved ELISA method for quantitation of and their products. Comparison and critical evaluation of six
aflatoxin Bx by utilization of second antibody bound to beads published extraction and cleanup procedures for aflatoxin Mx
and filtration plates. Aflatoxin Bx as low as 1 ppb in peanut in liquid milk were carried by Shepherd et al. (230B); a pre-
meal and 0.1 ppb in human blood was detected by the im- packed reversed-phase cartridge method was shown to be the
munoassay method with recoveries of >80%. most satisfactory in terms of speed, cost, and cleanliness of
Dimitrov et al. (64B) discussed the results of international the final residue. Stubblefield et al. (247B) reported the results
collaborative studies on the analysis of aflatoxin Bx, B2, Gx, of a collaborative study on rapid HPLC method for deter-
G2, and M in com flour, peanut meal, and powdered milk. The mination of aflatoxin Mx and M2 in artificially contaminated
use of one- and two-dimensional TLC and GC methods gave fluid milks. The reversed-phase LC method for Mx and M2
best results in terms of simplicity, precision, sensitivity, and has been adopted official first action by AOAC.
reproducibility. The occurence of aflatoxins in food and the A new hydroxyaflatoxin Bx: aflatoxin M4 was separated by
development of methods for their determination were re- Lafont et al. (146B) from mycotoxin fractions isolated from
viewed. Gilbert et al. (92B) surveyed aflatoxins in peanut liquid or powdered milk. Solvent extracts of milk were pu-
butters, nuts, and nut confectionary products by HPLC with rified by column chromatography and purified aflatoxins were
fluorescence detection. Results showed that 31 of 32 samples separated by TLC on silica gel. Aflatoxin M4 was quantitated
of major national brand-name products contained less than by visual comparison with standards under UV light or by
10 ppb aflatoxin Bx. Gulyas (102B) determined aflatoxins Bx, fluorodensitometry.
B2, Gx, G2, and Mx in peanut meal, corn flour, and milk powder Trichothecenes and methods for their determination were
by overpressure TLC of purified extracts using silica gel plates reviewed by Bonchev et al. (24B). Pohland et al. (202B)
and chloroform-ethyl acetate-THF (8:12:0.6) at a pressure published a review with 29 references on the analytical
of 0.5 MPa. Aflatoxins were quantitated by emmission chemistry of deoxynivalenol including GC, TLC, HPLC, and
fluorometry. Sensitivity of method for aflatoxins was ap- GC-MS determination of the mycotoxin in foods. Yoshizawa
proximately 0.25 ng and for Mx approximately 0.5 ng. ( 303B) in a review with 17 references discussed the charac-
Huang et al. (124B) separated aflatoxin Bx, B2, Gx, and G2 teristics of mycotoxins, food contamination with trichothec-
by RP-HPLC and determined them by using UV at 365 nm enes, and assay methods for mycotoxin determination in foods
with detection limit of 3.2-3.9 ng. Kawamura et al. (137B) and biological samples.
developed a sensitive method for the determination of afla- Bata et al. (11B) recommended a transesterification method
toxin Bx as its water adduct AFB2a in peanuts, cassava, and for rapid quantitation of trichothecene toxins in foods and
corn. The method used normal-phase and reversed-phase feeds, using both HPTLC and a capillary GC method. Blaas

216 R * ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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et al. (19B) described a GC-MS method for determination of procedure using Florisil and Sep-PAK columns. DON and
deoxynivalenol (DON) and nivalenol (NV) at the ppb con- NIV in the column eluates were quantitated by GC-ECD and
centration range. A liquid chromatographic cleanup step was GC-MS (single ion monitoring). Terhune et al. (261B) de-
followed by silylation of the extract and the resulting TMS- scribed a GC-ECD method for quantitation of DON in wheat,
ethers were determined by GC-MS monitoring the charac- corn, and feed at levels as low as 20 ppb.
teristic ions in the negative chemical ionization mode. Tiebach et al. (265B) determined DON and NIV in natu-
Chromatography was performed on a DB-5 fused silica ca- rally contaminated cereal samples by using direct liquid in-
pillary column at 80-280° with helium as carrier gas. Brumley troduction HPLC-mass spectrometry in a negative ion
et al. (32B) discussed negative ion (NI) mass spectrometry of chemical ionization mode and selected ion monitoring. HPLC
DON under resonance electron capture conditions. Certain was performed on a column of Fine Sil C18-10, with aceto-
fragmentations were studied by collision-induced decompo- nitrile-water (1:1) mobile phase Cereal samples were extracted
sition/mass-analyzed kinetic energy spectroscopy. The NI with methanol, defatted with hexane, and cleaned up on a
technique was applied to the identification of DON in extracts Florisil column. Confirmation of DON and NIV was also
of grains and snack foods by using on-column injection ca- performed by capillary GC-MS after silylation. Trenholm
pillary GC. The sample extract was obtained by extraction et al. (271B) evaluated different extraction procedures and
with acetonitrile-water and was subjected to TLC as a equipment for the analysis of naturally contaminated grain
quantitative screening or isolation step. products for vomitoxin analysis and reported on the merits
Cohen et al. (47B) described a comparative study between of different techniques. Trucksess et al. (273B) reported a
two Canadian methods on the analysis for vomitoxin in cereal modification of their TLC method (1984) for determination
grains and showed that the use of Sephadex LH-20 for pu- of DON in high-sugar breakfast cereals, corn syrup, and beer.
rification of extract along with a GC capillary system gives Moniliformin was determined by Jansen et al. (129B) in
better accuracy and sensitivity in analysis and prevents in- vegetable foods and feeds. The mycotoxin was extracted by
terference that occurs in packed column GC. A capillary Soxhlet extraction with methanol from moldy corn, rice, rye,
GC-EC determination of T-2 toxin, HT-2 toxin, and diacet- oats, wheat, and barley samples and was determined by TLC
oxyscirpenol (DAS) in cereal grains at levels as low as 100 ppb using MBTH as a new derivatizing agent for moniliformin.
for T-2 and DAS and 50 ppb for HT-2 was described by Cohen The moniliformin derivative was assayed at 518 nm. Scott
et al. (48B). The method was applied to wheat, oats, and et al. (219B) developed a rapid method for determination of
barley with average recoveries ranging from a low of 65% for Fusarin C in cereal products by RP-HPLC on normal-phase
T-2 in barley to a high of 99% for DAS in oats. Eppley et HPLC with UV detection at 365 nm. TLC and solvent sta-
al. (74B) reported on the results of a collaborative study on bility of moniliformin mycotoxin have also been investigated.
a TLC method for DON in winter wheat, the results of the Shepherd et al. (231B) determined moniliformin in maize
study showed that false positives were not a problem and that using ion-pairing extraction and ion-pairing HPLC with UV
all of the collaborators could detect DON at the 300 ppb level detection.
or higher. The method has been adopted official first action Carman et al. (37B) described a rapid method for the de-
by AO AC. termination and confirmation of the plant toxin myristicin
The application of online supercritical fluid extraction with in fresh, frozen, and canned carrots by GC Of purified extracts
chemical ionization (Cl) mass spectrometry (with ammonia on a nonpolar stationary phase (OV-101) using FID. Sus-
as the Cl reagent), and collision induced dissociation (with pected positive findings were confirmed by using a second
argon as target gas) tandem mass spectrometry, for the rapid stationary phase of moderate polarity (OV-225). Kellert et
identification of ppm levels of several trichothecene myco- al. (138B) determined Alternatia toxins in moldy tomatoes,
toxins in wheat samples was demonstrated by Kalinoski et apple preserves, fruit, and tomato juices, etc. by GC/MS.
al. (133B). Maycock et al. (164B) described the preparation Stack et al. (243B) described a liquid chromatographic method
of the p-nitrobenzoyl derivatives of the 4-trichothecene toxins for determining tenuazonic acid (TA) and alternariol methyl
T-2, diacetoxyscirpenol (DAS), nivalenol (NV), and deoxy- ether (AME) in tomatoes and tomato products, based on
nivelenol (DON) and their separation by HPLC with UV extraction of Alternaria metabolites from an aqueous slurry
detection at 254 nm. Vomitoxin was determined in grains and of the sample with chloroform, centrifugation, and fraction-
feed samples by Noonpugdee et al. (182B) by a combination ation on a silica gel column. Reversed-phase HPLC with UV
of semipreparative HPLC and capillary GC-ECD. The limit detection for TA at 280 nm and fluorescence detection for
of detection was 50 ppb in corn. Levels found in corn, barley, AME were used for final separation and determination.
oats, wheat, corn spoilage, and mixed feeds were given. Chu et al. (42B) developed an ELISA assay for the detection
Romer (213B) reported that small charcoal/alumina of saxitoxin in shellfish. PSP toxins in shellfish were deter-
cleanup columns were effectively used to remove interfering mined by Jonas-Davies et al. (132B) using an automated an-
materials from grain, feed, and food extracts prior to chro- alyzer continuous flow reaction system to oxidize toxins to
matographic determination of trichothecene mycotoxins. A derivatives which are detected by fluorescence, while Sullivan
TLC method (with ethyl acetate-methanol (20:1) as mobile et al. (248B) used an HPLC procedure for determining the
phase) has been developed that can simultaneously detect ppb toxins.
levels of DON, fusarenon X, nivalenol, T-2 toxin, HT-2 toxin, Kubacki (144B) published a review with 33 references on
neosolanical, and diacetoxyscirpenol in food and feed samples. methods for the determination of patulin in apple juice.
The use of charcoal/alumina cleanup columns in conjunction Special attention was paid to HPLC. Data on the occurrence
with HPLC and GC of trichothecenes was also discussed. A of patulin in apple juice and a few other apple products were
rapid method for detecting DON was developed by Shannon included. Experimental results on the role of technological
et al. (227B) who employed a C18 silica gel reversed-phase processes in decreasing patulin contamination of apple juice
column to remove impurities from grain extracts prior to TLC and apple wine were presented. Acar et al. (IB) reported on
screening of DON using aluminum chloride as spraying agent the occurrence and determination of patulin in commercial
to develop the blue response characteristic of the mycotoxin. tomato pastes. Patulin was extracted from tomato paste with
Total time involved was approximately 30 min and the method ethyl acetate, followed by cleanup on a Florisil column and
was applicable to corn, wheat, and barley at detection levels two-dimensional TLC using Silica Gel F254. The solvent
of 1 ppm and oats at 1.5 ppm, as well as environmental sam- systems were benzene-methanol-acetic acid (90:5:5) and
ples at 0.75 ppm. Steinmeyer et al. (245B) described a method toluene-ethyl acetate-formic acid (5:4:1). Spot visualization
for determination of deoxynivalenol and nivalenol in cereals was with 4% dianisidine and chlorine gas. The detection limit
by capillary GC-ECD of their heptafluorobutyrate derivatives was 40 ppb, and patulin was absent in 23 investigated tomato
DON and its metabolite DON-1 were analyzed in milk by paste samples.
Swanson et al. (249B) using C18 cartridges for purifying the A method for HPLC determination of patulin in fruit juices
analyte solution for subsequent preparation of the TMS-ether was reported by Ehlers (69B). Patulin was separated from
derivatives which were GC-ECD analyzed on an OV-17 col- hydroxymethylfurfural and determined by chromatography
umn. on a silica gel 40 column with toluene-ethyl acetate (3:1) and
Tanaka et al. (255B) developed a sensitive method for the then on a Sep-PAK cartridge with the same solvent. Eluted
simultaneous analysis of DON and nivalenol in cereals. The patulin was concentrated, redissolved in methanol-ethyl
toxins were extracted with acetonitrile-H20 (3:1), defatted acetate (9:1), and HPLC analyzed on a Lichrosorb RP-18 with
with hexane, and purified by a two-step chromatographic gradient elution and UV detection at 280 nm. Limit of de-
ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 217 R
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tection was 1 ppb and recovery from apple juice containing extraction and preliminary cleanup, the mycotoxins were easily
160 ppm was 80%. Eller et al. (73B) presented a procedure separated by TLC using toluene-ethyl acetate-90% formic
for the evaluation of patulin content of fruit and vegetable acid (6:3:1) developing solvent. The developed chromatogram
juices and purees: two-dimensional TLC using silufol plates was exposed to vapors of pyridine or acetic anhydride or a
with trichloroethane-acetone-80% formic acid (16:3:1) and mixture was overspotted on the mycotoxins. With this
toluene-ethyl acetate-86% formic acid (5:4:1), development treatment ochratoxin A, citrinin, penicillic acid, and zeara-
with benzidine HC1, and detection at 365 nm. Patulin was lenone were converted to new fluorescent compounds, which
found in apple powder (80 ppm), molded strawberry (32 ppb), were rechromatographed with the same developing solvent
and molded mandarin orange (10-80 ppb). and examined under UV at 365 nm. Sterigmatocystin was
Forbito et al. (80B) described a rapid method for the confirmed chemically using TLC plates impregnated with 0.6
quantitative determination of patulin in apple juice. The N sulfuric acid or 10% oxalic acid in methanol.
mycotoxin was extracted from the sample with ethyl acetate Lepom (148B) simultaneously determined citrinin and
and the extract was cleaned up by extraction with a sodium ochratoxin A in wheat and barley using RP-HPLC-fluores-
carbonate solution, and patulin was determined by RP-HPLC cence detection. Valente et al. (276B) developed a minicolumn
using a ^Bondapak C18 column and UV detection at 254 nm. screening and a TLC quantitation methods for determining
Ogawa et al. (184B) published a method on the determination ochratoxin A in corn, peanuts, beans, rice, and cassava. A
of penicillic acid and patulin in commercial fruit juice by GLC method for the determination of ochratoxin A in pork kidneys
of their trimethylsilyl derivatives. was developed by Wilken et al. (294B). Homogenized sample
TLC determination of sterigmatocystin in cheese was re- was extracted with ethyl acetate, acidified, and chromato-
ported by Francis et al. (81B). Cheese was extracted with graphed on a silica gel column eluting ochratoxin A with dilute
acetonitrile-4 % potassium chloride, the extract was parti- formic acid in chloroform. The evaporated eluate was re-
tioned with aqueous calcium chloride, hexane, and methylene dissolved in methanol and analyzed by two-dimensional TLC
chloride, and passed through a column of copper carbonate- with fluorescence detection or by RP-HPLC with fluorescence
diatomaceous earth (1:2) eluting with methylene chloride. The detection.
eluate was evaporated and sterigmatocystin determined by Several reviews were published related to pharmaceutical
chromatography on Sil-G-25 HR plates with benzene-meth- residues in food. Bishop et al. (18B) wrote a review with 59
anol-acetic acid (85:10:5) and was visualized by spraying the references on classical and recent methods for antibiotic de-
plate with A1C13. The fluorescence of the spot was enhanced termination in milk. A review by Bories (25B) discussed the
10-fold by additional spraying with a silicon-ether mixture control of pharmaceutical residues in foods of animal origin
enabling detection and quantitation at 2 and 5 ppb, respec- and methods of determining residues with the example of
tively. Average recoveries were 88.3% and 86.4% at the 10 anabolic hormones. Clark et al. (43B) discussed screening
and 25 ppb levels. methods and instrumental techniques for determining drug
Zearalenone and deoxynivalenol were determined by Ben- residues in foods of animal origin and the potential use of
nett et al. (15B) in corn, wheat, oats, rice, and barley. The enzyme immunoassay, and of two mass spectrometers in
toxins were extracted with aqueous methanol and partially tandem in screening for veterinary residues. Petz (200B)
purified by partitioning into ethyl acetate and then defatting reviewed chemical residue analysis of veterinary drugs in food,
with acetonitrile-petroleum ether. Toxins were then isolated general methodology, and GC procedures. Shaikh et al. (226B)
by silica gel column chromatography. Interfering materials published an overview of physical-chemical methods for de-
were removed from the column with benzene; zearalenone was termining aminoglycoside antibiotics in tissues and fluids of
eluted with benzene-acetone (95:5), and after a column wash food-producing animals.
with chloroform-methanol (98:2), deoxynivalenol was eluted Archimbault et al. (6B) determined chloramphenicol in
with chloroform-methanol (95:5) and zearalenone was quan- poultry by reversed-phase HPLC and absorbance detection.
titated by TLC and deoxynivalenol by gas chromatography Radioimmunological determination was used by Arnold et al.
of the TMS derivative. The detection limit for each toxin was (7B) for chloramphenicol residues in swine muscle, milk, and
about 0.02 ppm. Recoveries of added toxins varied with eggs. Bergner-Lang et al. (17B) determined chloramphenicol
substrate and level of toxin. in milk by reversed-phase HPLC-EC. Haagsma et al. (105B)
Bennett et al. (16B) reported on the results of a collaborative described a rapid sample preparation method for the deter-
study on an HPLC method for the determination of a-zear- mination of chloramphenicol in swine muscle by HPLC.
alenol and zearalenone in com. The method has been adopted Keukens et al. (139B) reported two HPLC methods for
official first action by AOAC. Czerwiecki (54B) determined analysis and confirmation of chloramphenicol residues in meat
zearalenone in wheat and rye samples by cleaning up grain with off-line cartridge sample cleanup and on-line diode array
chloroform extracts on silica gel column and separation of UV-Vis detection. Schmidt et al. (217B) used electrochemical
column eluate by TLC. The mycotoxin was detected by detection to determine chloramphenicol residues in fish.
spraying the plate with alcoholic aluminum chloride, heating Determination of diethylstilbesterol in canned steamed pork
at 130° for 5 min, and visualization under UV. Limit of by gas chromatography was reported by Lin et al. (149B). Van
detection was 300 ppb. Peteghem et al. (277B) reported a method for diethylstil-
Liu et al. (150B) developed a competitive indirect en- besterol residue determination in meat samples that involved
zyme-linked immunosorbent assay (ELISA) for the detection chromatographic purification and radioimmunoassay.
of zearalenone in methanol-water extracts of corn, wheat, and Ashworth (8B) reviewed the liquid chromatographic assay
pig feed samples. Warner et al. (288B) reported a screening of tetracyclines in meat. Botsoglou et al. (26B) determined
method for zearalenone in corn by competitive direct en- tetracycline, oxytetracycline, and chlorotetracycline by re-
zyme-linked immunosorbent assay. Average zearalenone re- versed-phase HPLC with monitoring at 361, 357, and 377 nm,
coveries from corn spiked at levels of 57, 151, and 307 ppb respectively. Hoshino et al. (121B) analyzed purified antibiotic
were 50, 119, and 123%, respectively. Levels of zearalenone extracts from beef, pork, and chicken using reversed-phase
detected in naturally contaminated samples were comparable HPLC and UV detection at 268 nm.
to those determined by liquid chromatography. Moats (167B) determined tetracycline antibiotics in blood
Tanaka et al. (254B) reported a rapid and sensitive method serum, muscles, liver, and kidney of cattle by HPLC employing
for the determination of zearalenone in cereals by HPLC with polymeric and reversed-phase column. Oka et al. (186B)
fluorescence detection. Zearalenone was extracted with ace- semiquantitatively detected and determined tetracycline and
tonitrile-water (3:1), purified on a Florisil column, resolved related compounds in foods by chromatography on high-
by using 90% water saturated chloroform-cyclohexane-ace- performance or reversed-phase silica gel TLC plates.
tonitrile-ethanol (50:15:2:1), and quantitated by fluorescence Prepacked C18 cartridges were used by Oka et al. (187B)
measurement detecting zearalenone in wheat, barley, corn, for tetracycline residue analysis in animal liver. Meat and
and cereals with picogram sensitivity. A combination of this fish were analyzed by Onji et al. (190B) for its tetracycline
HPLC method with GC method for trichothecenes may be content using HPLC-UV after sample cleanup on an Am-
applied to the simultaneous detection of zearalenone, niva- berlite XAD-2 column. Honey oxytetracycline residues were
lenol, and deoxynivalenol In cereals. analyzed by using reversed-phase HPLC-UV at 355 nm, by
Golinski et al. (94B) described chemical confirmatory tests Sporns et al. (241B) and by Takeba et al. (253B). Terada et
for ochnatoxin A, Citrinin, penicillic acid, sterigmetocystin, al. (258B) determined tetracycline residues in foods using
and zearalenone performed directly on TLC plates. After RP-HPLC-UV at 340 nm. Traldi et al. (270B) described a
218 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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rapid and sensitive method for the selective detection and parasitics, and growth promoters in meat, milk, and eggs with
quantitation of oxytetracycline residues at the ppb levels in emphasis on sulfonamide determination. Nagata et al. (174B,
crude extracts of bovine meat and milk. The technique em- 175B) described HPLC methods for the determination of
ployed was a tandem mass spectrometric method: CAD residual thiamphenicol and ethopabate in chicken. Neidert
MIKE spectrometry. Quantitation was performed by single et al. (181B) measured sulfathiazole residues in honey using
ion monitoring of CAD species of oxytetracycline and of its a rapid quantitative TLC and fluorescent scanning densito-
internal standard. Shaikh et al. (225B) described a method metry. Residual thiamphenicol in yellow tail muscle was
for the determination of neomycin in animal tissue. Heat determined by Otsuka et al. (192B) by RP-HPLC. Parks
deproteinized extracts of animal tissues were acidified to pH (196B) developed two procedures for the simultaneous de-
3.5-4 and directly analyzed by-RP-ion paired HPLC with termination of sulfa drugs and/or dinitrobenzamide coccid-
o-phthaldehyde postcolumn derivatization and fluorometric iostats, and their monoamine metabolites in chicken liver.
detection. Parks et al. (197B) quantitated zoalene and its two major
Lactam ring antibiotics in milk were detected by Brown et monoamino metabolites using RP-HPLC with an electro-
al. (31B) using a simple fast and inexpensive enzyme immu- chemical detector in the reductive mode. Paulson et al. (198B)
noassay. Moats (168B) determined penicillin G and cloxacillin described HPLC and GC-MS methods for identification and
residues in pork and beef tissues by RP-HPLC-UV at 220 nm. quantitation of sulfamethazine metabolites in swine tissues.
The method was suitable for other monobasic penicillins but Schwartz et al. (218B) described a simple method for rapidly
not for dibasic or amphoteric penicillins. Chromatographic screening sulfathiazole in honey. A GC-MS procedure using
methods were reported by Moats (169B) for the determination isotopically labeled internal standards and selective ion
of macrolide antibiotic residues in tissues and milk of food monitoring was used by Simpson et al. (237B) to assay five
producing animals. Two-dimensional TLC has been used for sulfonamide residues in liver and muscle tissue of swine,
the detection of tylosin in poultry, meat, eggs, and milk and poultry, and cattle.
for erythromycin in poultry meat, and HPLC using re- Tranquilizer residues, namely, xylazine, azaperone, acep-
versed-phase end-capped C18 columns which has also been romazine, propylporomazine, and chloropromazine, were de-
used for determination of tylosin in milk, blood, and tissue termined in alkaline homogenates of beef, pork meat, and
of animals. The LC method was more sensitive than micro- kidney by extraction into an organic solvent and then into
biological assays for detection of tylosin in tissues of treated an acid followed by RP-HPLC with UV and amperometric
swine, and recoveries of tylosin by LC method were frequently detection as reported by Etter et al. (75B). Blomkvist et al.
several-fold higher. Munns et al. (173B) reported a multir- (22B) reported on the qualitative and quantitative analysis
esidue method for the determination of eight neutral /3-lactam of monensin A sodium salt in the low nanogram range in
penicillins in milk by RP-HPLC-fluorescence detection. samples of chicken fat by TLC and fast atom bombardment
Nagata et al. (176B) used RP-HPLC-UV at 222 nm to de- MS. Blum et al. (23B) determined the thyrostatic drug me-
termine ampicillin residues in fish tissues. Horie et al. (119B) thylthiouracil (MTV) by HPLC and HPTLC in tissues of
determined spiramycin in swine, cattle, and chicken muscles frozen-thawed samples of fattening cattles. Hoshino et al.
using RP-HPLC-UV at 232 nm and identification by MS. (30B) determined monensin in chicken meat by HPLC with
Nagata et al. (177B) determined spiramycin in chicken tissue fluorescence detection. Monensin was also determined in milk
by using Zorbax C8 column-UV at 231 nm. Kanamycin was by Karkocha (135B) by two-dimensional TLC. TLC plates
determined by Nakaya (178B) in muscle and kidney of cattle sprayed with a 3% vanillin solution in methanol was used for
using RP-HPLC-fluorometry. visualization. Martinez et al. (159B) developed a method in
Terada et al. (259B) simultaneously determined penicillin which monensin residues were extracted from beef liver tissue,
G, penicillin V, and ampicillin in milk using RP-HPLC with acetylated, partitioned, and reacted with 9-anthryldiazo-
a mobile phase containing sodium alkylsulfonate. Prior to methane (ADAM) to form a fluorescent derivative for quan-
HPLC, samples were pretreated with a Sep-PAK C18 cartridge. titation by LC. A multiresidue method was developed by
The peaks corresponding to each /3-lactam antibiotic can be Martinez et al. (160B) whereby residues of monensin, sali-
confirmed with the treatment using penicillinase. Terada et nomycin, narasin, and lasalocid were determined in beef liver.
al. (260B) also determined penicillin G in edible animal tissues. The ionophores were extracted with aqueous methanol, pu-
The method consists of an off-line cleanup step using a basic rified by both alumina and Sephadex LH-20 column chro-
aluminum oxide column and a Sep-PAK C18 cartridge and an matography, and then derivatized with ADAM (directly with
on-line precolumn concentration and purification system. lasalocid) and via acetylation for the other three. The ADAM
Thorogood et al. (264B) reported on their evaluation of the derivatives were then purified on a silica gel column and
Penzim method (a rapid enzymic assay) for the detection of determined by RP-8 HPLC column using fluorescence de-
(3-lactam antibiotics within 20 min. Raw whole milk was used tection. Takatsuki et al. (251B) analyzed monensin residues
to establish its repeatability and reproducibility for the de- in chicken tissues by normal-phase HPLC of its ADAM
termination of penicillin G. Electrophoric methods for the fluorescent derivative.
detection of residual antibiotics in meat and eggs were re- Dimenna et al. developed a TLC bioautographic assay for
ported by Ionova et al. (127B) and in meats by Tao et al. satinomycin in chicken liver (62B) and in chicken fat and skin
(256B). (63B). Awasthi (9B) described an extraction cleanup method
Haagsma et al. (103B) described a rapid procedure for the for GC determination of synthetic pyrethroid residues in fruits
determination of sulfamethazine in swine tissues which com- and vegetables. Ito et al. (128B) determined natamycin, an
prises sonication and extraction with methylene chloride, antifungal residue, in natural cheese by UV spectrophotometry
cleanup of the extract on a combination of silica and re- and RP-HPLC.
versed-phase C18 Sep-PAK cartridges, and RP-HPLC analysis Aitzetmueller et al. (2B) developed a normal-phase HPLC
on a Hypersil ODS column with acetonitrile-10 mM ammo- method for determining metaldehyde residues in vegetables.
nium nitrate (3:1) at pH 6.8 with UV detection at 254 nm. Metaldehyde was extracted with toluene and then depolym-
Haagsma et al. (104B) reported a rapid method for deter- erized by acid and derivatized to acetaldehyde dinitro-
mining five sulfonamides in ground pork using a cation-ex- phenylhydrazone which was chromatographed and detected
change resin column for cleanup followed by analysis of pu- by UV. Bloeck et al. (21B) determined aldehydes and ketones
rified extracts on a Chrompack column and UV detection at in bottled and canned apple juice by converting them into
254 nm. The results of a collaborative study on a TLC corresponding 2,4-dinitrophenylhydrazones which were ana-
screening method for the detection of five sulfonamides in lyzed by reversed-phase HPLC on a Zorbax ODS column with
swine tissue were reported by Haagsma (106B). Hobson- 42% aqueous acetonitrile and absorbance monitoring at 365
Frohock (117B) used GC and HPLC for determining medicinal nm. Buckley et al. (33B) used electron capture gas chroma-
additives in poultry tissue. The use of droplet countercurrent tography to determine traces of formaldehyde in milk as its
chromatography and size exclusion chromatography was 2,4-dinitrophenylhydrazone. DeFreitas et al. (56B) determined
suggested to improve sample cleanup. Application of the formaldehyde in meat by photometry with acetylacetone.
photodiode array detector in the field of residue analysis was Hayashi et al. (110B) determined trace amounts of the mu-
discussed and an example given of its use in methodology tagen methylglyoxal in foods and beverages by a newly de-
development. Malisch (155B) described a procedure for the veloped method that involves reacting Me glyoxal with cys-
extraction, cleanup, derivatization, and quantitative deter- teamine at pH 6 to give 2-acetylthiazolidine which was ana-
mination (by GC and HPLC) of >60 antimicrobials, anti- lyzed by GC on a fused silica capillary column and monitored

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 219 R
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by thermionic N/P specific detector; formaldehyde was de- compared. Solvents released from printing inks and adhesives
termined in coffee as its thiazolidine derivative, by Hayashi can be distinguished from thermal decomposition products.
et al. (11 IB) as well as sweetened commercial food items Zenner et al. (305B) determined the volatile solvents given
(112B). Trace amounts of formaldehyde in aqueous extracts off by pack material coatings for food by headspace GC on
and model mixtures simulating foods were determined by a column of 20% polyethylene glycol adipate on porolith.
Pertsovkii et al. (199B) by GC of its 2,4-dinitrophenyl- Xylene, ethylene glycol, ethylene glycol acetate, acetone, ethyl
hydrazone derivative. The method was suitable for monitoring acetate, ethanol, butyl acetate, and toluene were found.
the migration of formaldehyde from polymeric materials into Results were discussed with respect to food contamination
food products. by these compounds.
Shin et al. (233B) analyzed n-hexanal in headspace vapor Tencheva et al. (257B) determined the concentration of free
over cooked brown rice by direct vapor injection GC-FID on radicals in irradiated prunes, nuts, and corn using EPR
a column of 18% Carbowax 20M on Chromosorb HP and spectroscopy. Ward et al. (286B) applied liquid chromatog-
correlated between oxidative deterioration of unsaturated raphy-electrochemical detection for the determination of toxic
lipids and n-hexanal during the storage of brown rice (224B). byproducts of boilable polyurethane cooking pouches.
Black et al. (20B) detected hydrogen peroxide in pasteurized Ethylene dibromide (EDB) residue confirmation in grain
cream by mixing the cream with tungstic acid, filtering, and and food products was reported by Barry et al. (10B) using
forming a stable yellow complex in the filtrate with titanium GC/MS with electron impact or negative ion chemical ioni-
tetrachloride the absorbance of which was measured at 415 zation selected-ion monitoring.
nm. Srinivas et al. (242B) described a method for determining The three commonly used methods for the determination
hydrogen peroxide in milk which is based on the catalytic of ethylene dibromide (EDB) fumigant residue in grains and
decomposition of H202 by a 7 irradiating catalyst and grain-based products, namely, soaking in hexane, triple co-
measuring the liberated oxygen with a gas measuring buret. distillation with hexane from an aqueous sample solution, and
A review on the contamination of foods by polycyclic aro- soaking in acetone-water (5:1), have been compared by Clower
matic hydrocarbons (PAH), polychlorinated biphenyls (PCBs), et al. (44B), and EDB recovery efficiency of each was reported.
and heavy metals was published by Gonzalez (96B) discussing Gilbert et al. (93B) developed an automated headspace ca-
levels, toxic effects, and determination of these contaminants. pillary GC-MS method using selected-ion monitoring for
Coates et al. (45B) analyzed PAH in plant materials using a determination of EDB residues in fresh fruits. Heikes (113B)
method that involved extraction with acetonitrile (sonication), developed a purge and trap method for determination of EDB
partition into pentane, and fractionation on a microsilicic acid in whole grains, milled grain products, intermediate grain-
column followed by GC-FID. Joe et al. (131B) determined based foods, and animal feeds and in table-ready foods (114B).
trace levels of carcinogenic and noncarcinogenic PAHs in EDB collected on adsorbent Tenax TA is eluted with hexane,
smoked foods, by reversed-phase HPLC on a Zorbax ODS determined by GC-EC, and confirmed with Hall electrolytic
column with both fluorescence and UV detection, PAHs were conductivity detection using a second GC column. The highest
determined by Lawrence et al. (147B) at nanogram/kilogram levels of EDB were also confirmed by full scan GC/MS.
levels in meat, fish, dried dairy products, cereals, leafy veg- Konishi et al. (142B) determined EDB in foods and grains by
etables, and vegetable/marine oils by reversed-phase gradient GC-EC using a high-resolution capillary column. Flour
elution HPLC and fluorescence detection. Takatsuki et al. products were GC analyzed for EDB by McKay (165B) using
(252B) presented a method for PAHs in fish and shellfish a 10% SP1200 + 1% phosphoric acid on Chromosorb W
using HPLC and fluorescence detection. Welling et al. (290B) column. Automated purge and trap GC-EC methods were
compared two cleanup procedures for analysis of PAHs in used by Moon et al. (170B) and by Pranoto-Soetardhi et al.
edible oils, one method comprised a liquid-liquid extraction (204B) for EDB residue analysis in rice and wheat flours and
followed by XAD-2 chromatography and the other involved cereals. Van Rillaer (278B) determined residual EDB in wheat
a complexation with caffeine-formic acid followed by HPLC cereals, and dried fruits by GC-EC.
in combination with wavelength programmed fluorescence DeVries et al. (58B) determined carbon tetrachloride,
detection. ethylene dichloride, and ethylene dibromide in grain and
Morozzi et al. (171B) described an analytical method for grain-based products by capillary GC-EC after hexane co-
determining a-benzopyrene and other PAHs in smoked or distillation using an internal standard (1,2-dichloropropane
cooked foods using TLC on cellulose acetate plates and de- and 1,2-dibromopropane). A headspace GC method for de-
tection by UV fluorescence. Xia et al. (296B) used paper termining methyl bromide in food ingredients was developed
chromatography followed by fluorometric measurements of by DeVries et al. (59B), and Delton et al. (60B) determined
a-benzopyrene of benzene eluates from paper chromatograms. carbon tetrachloride and ethylene dichloride in grain by
3,4-Benzopyrene was determined by Song et al. (239B) in volatilization in a hermatically sealed glass container followed
edible oils by caffeine complexation, followed by silica gel by headspace GC-FID.
column chromatography and TLC on acetyl cellulose plates Stijve et al. (246B) reported on the results of an interla-
with densitometric detection. boratory study for determination by GC-MS of 2-chloro-
Dennis et al. (61B) described a method for the analysis of ethanol (a residue arising from ethylene oxide fumigation) in
nitro-PAH in foods using a coupled capillary GC/thermal spices. Toyoda et al. (269B) determined minute quantities
energy analyzer. Joe et al. (130B) analyzed several smoked of chloroform in foods by GC-MS analysis of pentane extracts
foods for basic N-containing polynuclear aromatic hydro- of the steam distillates of fats, oils, Tofu, noodles, and bev-
carbons (NPAH) using a Vydac C18 column with aceto- erages. Phosphine residues in rice were determined spec-
nitrile-water (9:1) with UV and fluorescence detectors con- trophotometrically by Rangaswamy et al. (209B) and by
nected in series. Scudamore et al. (220B) who determined it in cereal grains
The migration of substances from packaging materials into and nuts using GC on a Poropak Q column with P-specific
foods and food models and methods for determining such thermionic detection.
migrants in foods was reviewed by Giacin (89B). Chernitsyna N-Nitroso compounds in foods was the subject of a review
et al. (40B) described a headspace GC-FID method for de- by Eisenbrand (70B) who covered the detection and deter-
termining the migration of organic substances from poly- mination of nitrosamines in foods as well as the problems of
ethylene and polypropylene used as food contacting materials. sample contamination and artifact formation. Scanlan et al.
BHT migration from polystyrene and polypropylene pack- (216B) gave an update of new analytical techniques for de-
aging materials into a fatty food model was determined by termining volatile and nonvolatile nitrosamines in foods and
Cweik et al. (53B) using GC-FID on a 10% OV-101 on Gas- beverages and Sen (223B) wrote an extensive review on recent
Chrom Q column with argon as carrier gas. Acetaldehyde trends in nitrosamine analysis in foods.
migration from polyethylene terephthalate containers was Sen et al. (222B) developed a simple direct extraction
determined by Lorusso et al. (151B) using headspace GC-MS. method for the determination of volatile N-nitrosamines in
Snyder et al. (238B) evaluated a new FDA migration cell used rubber nipples and pacifiers. It consists of overnight extraction
to study migration of styrene from polystyrene into various of the sample with dichloromethane in the presence of ascorbyl
solvents. palmitate for N-nitrosation inhibition, filtration, concentration
Headspace capillary GC analysis of residual solvents in food using a Kuderna-Danish concentrator, and final analysis by
packages was applied by Rochelli et al. (212B). Two vapor- GC-thermal energy analyzer (TEA). Thompson et al. (263B)
phase injection systems on packed and capillary columns were determined N-nitrosamines and N-nitrosamine precursors in

220 R» ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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rubber nipples from baby pacifiers by GC-TEA. was determined colorimetrically with enzymic treatment.
Beattie et al. (13B) determined nonvolatile nitrosamines Histamine and agmatine were analyzed by Matsunaga et
by moving belt liquid chromatography/mass spectrometry al. (162B) by GC-MS. Matsunaga et al. (163B) simultaneously
(LC/MS) and spectra achieved by ammonia chemical ioni- determined tyranine, histamine, putrescine, /3-phenylamine,
zation. Dunn et al. (67B) analyzed cured meat samples for cadaverine, agmatine, and tryptamine in soy sauce, miso, and
N-nitrosoproline (NPRO) using methanolic extraction, fol- cheese by reversed-phase HPLC. Offizorz et al. (183B) de-
lowed by GC-TEA. Gavinelli et al. (88B) analyzed volatile termined histamine in fish and canned fish by capillary iso-
nitrosamines in foods by a new method involving simultaneous tachophoresis. Ramantanis et al. (207B) fluorometrically
distillation-extraction. Hu et al. (123B) determined volatile determined histamine and other biogenic amines in dry sau-
nitrosamines in foods by GC-TEA. The sample is distilled sage after extraction with TCA, followed by purifying the
at 110-120°, the distillate was extracted with dichloromethane, extract on a weak acid cation-exchanger, derivatization with
extract concentrated, and subsequently GC analyzed on a 10% o-phthaldialdehyde, and measurement of reaction products
polyethylene glycol) column. at 445 nm after excitation at 360 nm. This method was more
Gracheva et al. (98B) determined AT-nitrosamines by iso- precise than the TLC method by same authors (208B). Tiecco
lation from food samples, denitrosation, refluxing the sec- et al. (266B) used high-resolution liquid chromatography to
ondary amine obtained with an 8-10-fold excess of 8-meth- determine histamine and 10 other biogenic amines in sausages.
oxy-5-quinolinesulfonyl aziridine in aqueous alcohol at pH 7-9, Walters (285B) determined histamine and other related
and measuring the fluorescent derivatives formed. compounds in fish by HPLC with postcolumn reaction and
HPLC was applied by Kim et al. (MOB) for the determi- fluorometric detection.
nation of iV-nitrosodialkylamines in fermented and salted Hashiba et al. (108B) determined /3-phenylethylamine and
foods. Massey et al. (161B) determined AT-nitrosoamino acids putrescine in soy sauce by ion exchange chromatography.
by denitrosation/TEA. Sen et al. (221B) developed a simple Ingles et al. (126B) analyzed foods for their content of biogenic
amines by ion exchange and HPLC methods. In the latter
HPLC/chemiluminescence detection method for the analysis
of AT-mtrosamides in foods and beverages based on postcolumn the amines were chromatographed as their fluorescamine
denitrosation of the compounds with HBr-HOAC or HI- derivatives followed by identity confirmation using field-de-
HOAC, followed by the detection of liberated NO by a sorption MS.
chemiluminescence detector. Tonogai et al. (268B) developed a headspace GC method
AT-Nitrosothiazolidine and nonvolatile (iV-nitrosourea) in for the determination of volatile amines and ammonia in raw
foods were determined also by Sen et al. (224B) using the fish and fish products. Wakabayashi et al. (282B) quantified
chemical denitrosation/TEA method; in addition, two GC/ mutagenic and carcinogenic heterocyclic amines in cooked
TEA for determining nitrosothiazolidine and N-nitroso- foods by HPLC with a combination of reversed-phase and
thiazolidine-4-carboxylic acid in cereal, meats, and fish were cation-exchange columns with electrochemical detection.
mentioned. Piasecka et al. (20IB) reported an improved Yamaizumi et al. (297B) employed stable isotope dilution for
method for determining AT-nitrosamines in beef meat by determining mutagens in cooked food by combined LC-
isolation by distillation under reduced pressure followed by thermospray MS. Yen (300B) determined biogenic amines
GC on a column of 10% PEP 20M on Chromosorb AWDMCS. in fermented soybean by reversed-phase HPLC of their dansyl
Tricker et al. (272B) developed a method for the analysis of derivatives on a Lichrosorb RP-18 column with acetonitrile-
three AT-nitrosodipeptide derivatives in cured meat by HPLC methanol-H20 (1:2:1) mobile phase. An automated ion-ex-
on a Zorbax CN column, with acetone-acetic acid-hexane change chromatographic method for the composite analysis
(18:1:81) as mobile phase and chemiluminescence detection. of biogenic amines in cheese was used by Zee et al. (304B).
Ergot alkaloids in wheat and rye flours and breads and other Cuq et al. (52B) published a review on the formation of ly-
cereal products were determined by Baumann et al. (12B) sinoalanine in thermal processing of food proteins, especially
using reversed-phase HPLC-fluorometry after initial ex- at alkaline pH, and its determination in food, and Edmonds
traction and cleanup of samples by an extralut column. Klug et al. (68B) used directly combined HPLC/MS for the analysis
et al. (141B) described a method for determining ergometrine, of heterocyclic aromatic mutagens in cooked foods.
ergometrinine, ergosine, and ergotamine on a Hypersil ODS
column (3 jtm) with acetonitrile-water (47:53) as mobile phase. CARBOHYDRATES
Ware et al. (287B) determined ergot alkaloids in wheat by Several books have been published during this review period
HPLC using a porous cross-linked poly(styrene-divinyl- including: The Analysis of Food Carbohydrates, edited by
benzene) column with a mobile phase of acetonitrile-0.05 M Birch (17C); Carbohydrates, edited by Collins (21C); The
dibasic ammonium phosphate (55:45) at pH 10 and fluores- proceedings of the conference held in Copenhagen, Denmark,
cence detection. on “New Approaches to Research on Cereal Carbohydrates,
Potato glycoalkaloids were determined by Hellenaes (116B) edited by Hill et al. (37C); and the 42nd volume in Advances
who used Sep-PAK cartridges replacing commonly used al- in Carbohydrate Chemistry and Biochemistry, edited by
kaline precipitation for cleanup of tuber extracts, followed by Pigman et al. (64C).
reversed-phase HPLC and UV detection. Results showed good Several review articles have been published on carbohydrate
agreement between this method and an ELISA and colori- analysis. Clode (20C) discussed general methods of carboh-
metric methods. ydrate isolation and X-ray crystallography, TLC, GC, HPLC,
Colchicine alkaloids in tissues and biological fluids of cows IR, and NMR spectrometric, and polarimetric analysis of
were determined by Yoneda et al. (302B). Samples were carbohydrates. The analysis of food glycosides was reviewed
extracted with acetone, acetone solutions defatted with pe- by Dziedzic et al. (24C). Folkes et al. (28C) wrote a review
troleum ether, zinc sulfate, and barium hydroxide solutions on analytical methods for the characterization of glucose
added to the acetone extract for protein removal followed by syrups. Folkes (29C) reviewed GC methods for carbohydrate
chloroform extraction of alkaloids and subsequent HPLC-UV analysis in foods. Physical, chemical, and biochemical
at 350 nm using a column of Lichrosorb RP-18 with aceto- methods of analysis of food carbohydrates were reviewed by
nitrile-methanol phosphate buffer as mobile phase. Kearsley (41C). HPLC methods for determining sugars and
Cyanogenic glycosides and cyanohydrins were determined polysaccharides in food including information on extraction,
by Brimer et al. (30B) in plant tissues by enzymic postcolumn cleanup, separation, and detection systems, as well as ap-
cleavage and electrochemical detection after reversed-phase plications were covered in a review by Macrae (50C). Pirisino
HPLC. Chikamota et al. (4IB) indirectly determined cyanide (65C) reviewed fixed-ion resin and amino-bonded silica col-
concentrations in beans by measurement of acetone and 2- umns used in HPLC of carbohydrates in foods.
butanone formed from the hydrolysis of bean cyanogenic Methods of determining the degree of starch damage in
glycosides using GC-MS. Dalgaard et al. (55B) analyzed crude cereals including microscopic, enzymic, and instrumental
and partially purified extracts from cassava for cyanogenic procedures were published by Podgorska (66C). A review on
glycosides by electrochemical detection of cyanide ion liberated the application of gel permeation chromatography in the
by enzymic postcolumn cleavage. Detection limit was in the separation of starch components and derivatives from food
low picomole range. Maitani et al. (154B) determined cyan- was published by Pranzik (67C). Rathbone (68C) discussed
ogenic compounds in health foods made of Japanese apricot the use of NMR in the structural analysis of food-related
by treatment with /3-glucosidase followed by steam distillation carbohydrates. Reimerdes et al. (70C) reviewed qualitative
and colorimetry of liberated cyanide ion. Free cyanide content and quantitative chromatographic, spectrometric, and other

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 221 R
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methods of carbohydrate analysis in food. Shallenberger (79C) by the Swiss Official Methods Committee.
reviewed the principles of optical polarimetry, the optical Beach et al. (<SC) used a paper chromatographic method for
rotatory characteristics of sugars, and applications of polar- the separation and quantitation of minor nonabsorbed sugar
imetry to sugar structure studies and quantitative analysis (lactulose) and nonabsorbable oligosaccharides. Betschart et
in foods. Sumida et al. (83C) wrote a review on chromato- al. (9C) analyzed lactose hydrolysis products by HPLC on a
graphic analysis of sugars (mono-, di-, oligo-, and poly- sugar PAK-1 column with water as mobile phase. Cai et al.
saccharides) of soybean milk by HPLC. Q8C) determined mono-, di-, tri- and polysaccharides in honey,
Athnasios (4C) developed a method for the analysis of trace beets, grapes, sweet wine, and wort by HPLC on a
levels of D-glucosone in complex food matrices by isolating aBondapak/carbohydrate column with aqueous acetonitrile
D-glucosone using reversed-phase microcolumn chromatog- as mobile pnase. A Hypersil 5 amino column with aqueous
raphy, followed by reacting it with 2,4-dinitrophenylhydrazine acetonitrile, and infrared (IR) detector were used by Cirilli
(2,4-DNPH) to yield the corresponding osazone which was et al. (19C) for sugar determination in molasses, honey, fruit
subsequently determined by normal-phase TLC and nor- juices, and other food substances.
mal-phase HPLC, with absorbance detection at 436 nm. Den Deifel (22C) reported a TMS-GC-FID method for the de-
Drijver et al. (23C) used a strongly basic anion-exchange termination of sugars in honey. Goetz (34C) determined
HPLC column coupled with an amperometric detector to carbohydrates in dietetic nonalcoholic beverages by HPLC
detect D-glucosone in 7-irradiated sugar solutions, irradiated on an HPX-87A cation-exchange column with 0.01 N sulfuric
model fruit systems, as well as in irradiated whole fruit. acid as mobile phase; glucose at low concentrations could be
Enzymic-spectrophotometric methods for the analysis of analyzed in the presence of 250-fold fructose concentrations.
mono, di-, tri-, and polysaccharides as well as sugar alcohols Grossi et al. (35C) reported on results of comparative deter-
in food products were reported by Beutler who determined minations of sugar in wheat flour by GLC and HPLC and
xylitol (11C), D-sorbitol (12C), maltose (13C), raffinose (14C), indicated close correspondence between results of methods.
lactose and D-galactose (15C), and starch (16C). Fozy et al. Hughes et al. (39C) used HPLC to determine fructose, glucose,
(30C) determined glucose, lactose, and sucrose by an enzymic and sucrose in cabbage as well as residual sugars and mannitol
method in natural and milk chocolates. Frank et al. (31C) in sauerkraut.
developed a method for determining the lactose and sucrose Senkalszky et al. (78C) described a TLC method for the
contents of ice cream mix. The method is based on measuring determination of sucrose, raffinose, and stachyose in legu-
the freezing point depression resulting from incubating sam- minous plants. The sugars were extracted by refluxing with
ples with invertase and /3-galactosidase. Kleyn (42C) reported ethanol and were separated on Polygram cel 300 layers using
the results of a collaborative study of a method for enzymic propanol-ethyl acetate-water (6:1:3) as mobile phase.
determination of lactose in milk. The method was adopted Quantitation was by densitometry, following spot visualization
official first action by AO AC. with a-naphthol in phosphoric acid. Knudsen (43C) developed
Matsumoto et al. (52C) developed an amperometric flow an HPLC method for the determination of sucrose, raffinose,
injection method for the determination of fructose in fruits and stachyose in leguminous seeds as soybeans, chick peas,
using an immobilized fructose-5-dehydrogenase reactor. The garden peas, and red kidney peas using a LiChrosorb NH2
same authors (53C) determined lactose in milk by flow in- column, a 35% water in acetonitrile as mobile phase, and as
jection analysis using a chemically modified lactose electrode. interference type refractive index (RI) detector capable of
Lactose was determined in whey by hydrolysis with /3-galac- detecting as low as 10 ng of oligosaccharides. Kobayashi et
tosidase followed by oxidation of the galactose with galactose al. (44C) separated amylose and amylopectin components of
dehydrogenase, and subsequent spectrophotometric mea- starch by size-exclusion HPLC on a two-column system with
surement of the NADH formed, as reported by Reimerders dimethyl sulfoxide as mobile phase. The method proved
et al. (69C). Tsujisawa et al. (87C) determined lactose in milk, useful to monitor the purity of amylose and amylopectin
margarine, butter, cheese, ice milk, and milk powder by an preparations and to rapidly estimate the amylose/amylopectin
improved enzyme assay with a reagent containing d-galacto- ratio of starch samples. He et al. (36C) determined amylose
sidase, hexokinase, glucose-6-phosphate dehydrogenase, and in defatted rice grains by amperometric titration. Glucose
NADP in pH 7.5 phosphate buffer, followed by spectropho- and sucrose in soft drinks, breakfast cereal, and cake mix were
tometric measurement of NADH formed at 340 nm. determined by Koemer et al. (45C) by chemiluminescence flow
Miwa et al. (56C) reported an enzymic method for the injection analysis using enzymatic conversion and a micro-
simultaneous determination of glucose, fructose, and sucrose porous membrane flow cell.
in foods. After glucose was measured by hexokinase and Kunerth et al. (46C) modified the anthorne, carbazole, and
glucose-6-phosphate dehydrogenase, fructose was measured orcinol reactions used for the quantitation of hexoses, uronic
by the addition of glucose phosphate isomerase, and sucrose acids, and pentoses, respectively, to improve sensitivity in the
was successively measured by the aid of invertase. Walter et analysis of unavailable carbohydrate (fiber). Lercker et al.
al. (93C) reported on the results of a collaborative study for (49C) analyzed royal jelly for its content of glucose, fructose,
the enzymic determination of glycerol in grape juice, wine, and sucrose by high-resolution gas chromatography (HRGC)
and gum drops. The method involved the use of glycerokinase, of the sugar-TMS derivatives on a short capillary column with
pyruvate kinase, and lactate dehydrogenase with appropriate SE-52 stationary phase film and helium as carrier gas. Mardal
substrates and the formed NAD was measured spectropho- et al. (51C) developed a procedure for the determination of
tometrically. A glucose sensor was developed and applied by sugar content in beets using a nontoxic clarifying agent
Watanabe et al. (95C) for the determination of glucose in fish (calcium oxide and aluminum oxide) instead of lead acetate.
muscle and blood serum. The sugars and organic acids in the pulp of Keitt mangoes
Starch was determined in cereal grains by Aaman et al. (1C) at various stages of ripeness were analyzed by HPLC as was
using an enzymic method which includes gelatinization of reported by Medlicott et al. (54C). The major sugars were
starch and simultaneous partial hydrolysis using a ther- identified as glucose, fructose, and sucrose.
mostable a-amylase, complete hydrolysis using amylo- Molnar-Perl et al. (57C) analyzed soybean soluble sac-
glucosidase, and determination of released glucose by the charides up to pentasaccharides by TMS-GC. Nebytov et al.
glucose oxidase method. Garcia et al. (32C) compared the (58C) determined 14 sugars in sweet pepper, plum, tomato,
Fehling method and an enzymic method (using amino- and strawberry. Extraction of sugars with water was followed
glucosidase and glucose oxidase/peroxidase) for starch analysis by HPLC analysis using a hydoxylated silica gel column with
in meat products (sausage). Both methods were equally ac- adsorption modified by piperazine, aqueous acetonitrile as
curate but the specific enzyme method gave greater precision. mobile phase, and an interference refractive index detector.
The Fehling method, as adopted in official regulation, was Ohtsuki et al. (59C) determined free sugars in the hydro-
not useful for analysis of samples with less than 2 % starch. lyzates of water-insoluble substances of orange juice, brown
Holm et al. (38.C) enzymically determined starch in raw wheat sugar, rice bran, jam, roasted soybean flour, adzuki paste, and
and cereals. A method was described by Karkalas (40C) for corn meal and glucose, galactose, and fructose in kiwi fruit
the determination of native and modified starch in a variety (60C) by HPLC using an anion-exchange column with 0.2 M
of foods, and Yasui (100C) determined maize starch in glu- borate buffer (pH 8.4) and 0.7 M borate buffer (pH 8.7) for
tinous rice cake by MS 13C/12C isotope ratio. Walter (92C) stepwise elution of disaccharide and monosaccharides, re-
reported the results of collaborative study for the enzymic spectively, and by postcolumn reaction with 2-cyanoacetamide
determination of starch in foods; the method has been adopted at 280 nm.

222 R « ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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The determination of carbohydrates in food by ion exchange subsequent enzymic determination of liberated galactose.
HPLC of their borate complexes and postcolumn reaction was Bettler et al. (10C) determined mixtures of gelling and
also discussed by Reimerders et al. (71C). The same authors thickening agents in foods by electrophoresis on silylated glass
(72C) described an HPLC method for the analysis of lactulose fiber paper.
and epilactose in milk and milk products after heat treatment A gas chromatographic method was evaluated by Lawrence
in the presence of high levels of lactose and other sugars. The et al. (48C) for the determination of food grade gums in dairy
method is based on the separation of the sugar borate com- products, salad dressings, and meat sauces. Scherz (75C)
plexes by anion-exchange chromatography using two optim- described a qualitative and semiquantitative method for de-
ized buffer systems. termining alginates in foods whereby the polysaccharide is
Ion exchange HPLC on sugar-PAKl column and dis- hydrolyzed by methanolic HC1 and hydrolysis products were
tilled-deionized water containing 0.1% calcium acetate as separated by TLC and visualized by the uronic acid specific
mobile phase was employed by Melton et al. (55C) for the naphthoresorcinol-HCl reagent. The same author (76C, 77C)
analysis of sugars in apple juice. separated polysaccharide thickening agents by thin-layer
Ruggeri et al. (73C) determined sugars, particularly lactose electrophoresis on plates coated with octanol and borate
in human milk substitutes, by defatting with hexane, cen- buffer.
trifuging, filtering, and chromatographing an aliquot of the Sjoberg et al. (82C) reported a method for the identification
aqueous phase on a Micropak NH2-10 column with aceto- of food thickeners by pyrolysis GC and capillary GC/MS.
nitrile-water (80:20). Valdehita et al. (89C) separated sugars Thier (86Q identified and quantitated natural polysaccharides
from milk preparations by TLC and determined them by in in foodstuffs by capillary gas chromatography of methyl-
situ densitometry. The results of a collaborative study on an glycoside silyl derivatives.
HPLC method for the determination of lactose purity using Englyst et al. (25C) determined non-starch polysaccharides
fructose as an internal standard were reported by Saucerman in cereal produts by GLC of constituent sugars. Faulks et al.
et al. (74C). The method has been adopted official first action (26C) described a procedure for the determination of non-
by AOAC. Shidlovskii et al. (80C) colorimetrically determined starch polysaccharides in foods whereby starch is removed by
lactose content in sodium caseinate by precipitating casein treatment with heat-stable a-amylase and resistant starch by
with acetic acid in the presence of sodium acetate, reacting treating with dimethyl sulfoxide and amyloglucosidase fol-
casein-free filtrate with phenol, and measuring the phenol- lowed by colorimetric measurement of total neutral sugars and
lactose complex at 490 nm. uronic acids in the acid hydrolyzates of the starch-free ma-
Simon et al. (81C) evaluated 18 methods to quantitate terial.
reducing sugars in carrots by paper chromatography; reducing Forni et al. (29C) reported a method for determining the
sugar determination with dinitrosalicylic acid was rapid and methoxy number of pectins. Pectin was treated with NaOH
correlated very well with values obtained by HPLC. An au- and the liberated methanol is GC-FID using a Porapak Q
tomated colorimetric method for the determination of glucose, column packing and N as carrier gas. Petrzika et al. (62C)
fructose, and sucrose in fruits, vegetables, and fermentation reported a method for quantitative determination of the
liquors was described by Tawfik et al. (85C). The method uses monomers of pectins. Various pectin preparation were hy-
p-hydroxybenzoic acid hydrazide and 2-thiobarbituric acid drolyzed and trimethylsilyated and monomer derivatives
as color reagents for the determination of total reducing sugars analyzed by GC-FID on a column of 3% OV-17 on Chromo-
and fructose, respectively. Zheng et al. (101C) partially pu- sorb WHP using phenyl-/3-D-glucopyranoside as internal
rified reducing sugars of soy sauce by using acidic alumina standard. Voragen et al. (91C) investigated the analysis of
column chromatography, diluting with water, and determining pectins as the anhydrogalacturonic acid (AGA) content by
them by measuring color produced by reaction with 3,5-din- ion-pair HPLC of the degradation products resulting from
itrosalicylic acid at 475 nm. enzymic hydrolysis. Wedlock et al. (97C) reported a new
Xie et al. (99C) used 3-hydroxy-2-naphthoic hydrazide as colorimetric assay for pectins using 2-thiobarbituric acid as
a precolumn UV derivatizing reagent for the determination color reagent.
of sugars in honey by HPLC on an amino-bonded column Wang et al. (94C) examined the chemical method for the
using aqueous methanol as mobile phase. The carbohydrate estimation of available carbohydrates in foods from plants.
composition of tea plant honey samples was determined by The results were comparable with the bioassay method.
Zhu et al. (102C) using HPLC, GC, activated charcoal column Wenlock et al. (98C) reported an improved fractionation
chromatography, paper chromatography, and mass spec- method for determining dietary fiber values using a mixture
trometry. Paynter et al. (61C) evaluated an ion exchange of amyloglucosidase and a-amylase to digest starch; the fiber
HPLC system for measuring glucose content in foods, bev- components found in 138 cereal and cereal-containing meat
erages, bakery waste, and waste syrup by comparing the data products were given.
with results form the glucose oxidase assay system. Sugars Baker (5C) used near-IR reflectance spectroscopy to de-
in raw and baked potato were determined by Picha (63C) termine fiber, starch, and total carbohydrates in potato chips,
employing an ion exchange HPLC method with RI detector. corn chips, extruded snacks, popcorn, cracker, and pretzels,
U et al. (88C) determined sugars in mandarin juice by while Baker et al. (6C) used two computerized scanning
fractionating sample extracts using C18 Sep-PAK cartridges near-IR reflectance instruments to determine total sugar
followed by chromatography on a /uBondapak amine column content in ready-to-eat breakfast cereals.
using aqueous acetonitrile as mobile phase and detection by Near-IR spectrometry was used by Giangiacomo et al. (33C)
RI. HPLC on mixed cation-exchange resins was employed to determine glucose, fructose, and sucrose in aqueous mix-
by Van Riel et al. (90C) for the separation of mono-, di-, and tures at 10, 25, and 40% total sugars. Lanza et al. (47C)
trisaccharides mixtures. Watanabe (95C) described an HPLC applied near-lR spectroscopy in the transmission mode to
method for determining reducing carbohydrates in fruit juices predict the total sugar content of a variety of juices.
by an amino-bonded column with aqueous acetonitrile as
mobile phase and amperometric detection using copper bis- COLOR
(phenanthroline) as postcolumn reagent. Tamate et al. (84C) Amakawa et al. (ID) partitioned oil-soluble natural dye
determined glucose, fructose, sucrose, maltose, and raffinose extracts between hexane and ammonium hydroxide. The
in sweet potato and taro by 13C NMR and HPLC. hexane fraction was analyzed for /3-carotene content using
Sugars in hydrolyzed commercial food gums were deter- normal-phase HPLC, and the aqueous fraction components,
mined by TMS-GC using /3-D-glucopyranoside as an internal curcumin, norbixin, and bixin, were resolved on a Zorbax ODS
standard as reported by AL-Hamzi et al. (2C). Angelini et column. Ushiyama et al. (11D) fractionated natural oil-soluble
al. (3C) detected, identified, and determined some vegetable dye extracts from prepared foods using silica gel chroma-
gums, by hydrolyzing with 2 N trifluoroacetic acid in 80% tography and further resolved the three fractions by TLC.
ethanol and reducing the liberated monosaccharides with /3-Carotene, capsaicin, bixin, curcumin, chlorophyll, Cu
sodium borohydride to the corresponding sugar alcohols which chlorophyll, norbixin, Na Cu-chlorophyllin, and Na Fe-
were determined by GC of their acetate derivatives on a chlorophyllin were detected with TLC. Synthetic dyes were
column of 3% SP 2330 on Supelcoport. A method for the extracted from soft drinks and lemonade syrups using the
rapid determination of galactose containing thickeners in meat ion-pairing agent trioctylamine by Puttemans et al. (9D).
products was reported by Bauer et al. (7C) which is based on After back extration into aqueous sodium chlorate, the sam-
acid hydrolysis of samples under increased pressure and ples were analyzed by reverse-phase HPLC. The artificial dyes

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 223 R
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Amaranth, carmine, Lemon yellow, Sunset yellow, indigo, and methylumbelliferyl esters as substrates was developed for the
Brilliant Blue were similarly analyzed using reverse-phase determination of low lipase activities in foods by Haslbeck
HPLC by Ye et al. (13D). Nanogram detection was achieved et al. (14E). Linfield et al. (18E) developed a new method
by using 190 nm. Kamikura et al. (4D) analyzed the natural for lipase activity using olive oil as the substrate with periodic
colors crocin and crocetin in gardenia fruit by reverse-phase sonication. The determination of lipase activity and free fatty
HPLC. A variety of natural pigments and synthetic dyes acids of milk using colorimetric automatic titration was pro-
including carmine, amaranth, Sunset yellow, Brilliant Blue, posed by Cartier et al. (5E). McKellar and Cholette (20E)
indigo monascus color, safflower yellow, betainin, and laccaic modified the method for determining the extracellular lipases
acid were resolved and quantitated by TLC on polyamide and of Pseudomonas fluorescens based on hydrolysis of /3-naphthyl
polyamide-silica gel G mixtures by Yang et al. (12D). Kanda caprylate and photometry and applied it to skim milk. A
(5D) adsorbed water-soluble natural pigments on polyamide, method for lipase activity from P. fluorescens was optimized
then further resolved the compounds by TLC on polyamide by Paquette and McKellar (24E) using a super/simplex op-
plates. The pigments analyzed included enocyanin, safflower timization computer program. Stead (26E) developed a
yellow, lac dye, cochineal, betarin, gamboge, Cu Na chloro- fluorometric method for the determination of P. fluorescens
phyllinate, and Monascus dye. A method for quantitative lipase using the fluogenic substrate 4-methylumbelliferyl oleate
extraction of synthetic water soluble dyes from aqueous and adapted it for skim milk powder, whey powder, and whey
sources with 0.1 M quinine in chloroform in the presence of protein concentrate. A fluorometric assay of lipase in rice bran
0.5 M boric acid was developed by Kobayashi et al. (7D). was developed by Saunders and Heltved (25E) using 4-
Twenty-one dyes, including 11 azo, 5 triphenylmethane, 4 methylumbelliferyl heptanoate as substrate and applied to
xanthene, and 1 naphthol derivative, were tested. Yin et al. determination of conditions for rice bran stabilization.
(14D) resolved water-soluble food dyes by ion-pair reverse- Fretzdorff (HE) developed a method to determine lip-
phase TLC using silica gel G plates treated with phenyltri- oxygenase activity in corn, wheat, barley, and oats using an
chlorosilane and several ion pair reagents: Me4NBr, Et4NBr, O-selective electrode to measure the 0 uptake during the
Bu4NCl, or hexadecylbromopyridine. reaction.
Khachik et al. (6D) separated three classes of natural pig- Kwee (17E) described an improved method to determine
ments, xarithophylls, chlorophylls, and carotenoids, from alkaline phosphatase based on a photometric method which
vegetables by using reverse-phase HPLC. Cis-trans isomers is applicable to colored products and a wider variety of milk
and other within-class components were resolved. /3-Carotene, products. Methods of sample preparation for detecting al-
xanthophylls, and pheophytins were detected by spectrometry kaline phosphatase in casein were studied collaboratively by
at 451, 447, and 667 nm, respectively, after fractional isolation Murthy and Peeler (21E)\ the alternative rapid sample
on a polyamide column by Mader and Chladova (8D). preparation method was adopted official first action by the
Chlorophyll a and b, and their degradation products pheo- AOAC. Paggi et al. (22E) developed a colorimetric method
phytin a and b, were determined in vegetable oil using a for the determination of peroxidase in milk using H202 and
^Porasil column by Rahmani et al. (10D). Anthocyanins in the Rothenfusser reagent. Wasserman and Wagner (29E)
colored beverages were determined by sorbent extraction, described a rapid colorimetric peroxidase assay to determine
separation on cellulose TLC plates, and color reaction visu- blanch efficacy in green beans.
alization by Zloch (15D). A method to determine proteinase from P. fluorescens Pi
Aromatic amines, including benzidine, aniline, 4-amino- by an enzyme-linked immunoassay technique was described
biphenyl, and 4-aminoazobenzene, were detected and quan- by Birkeland et al. (2E) and it was found to offer the com-
titated in FD&C Yellow No. 5 (tartrazine) by Bailey and bination of sensitivity and specificity for the detection of these
Bailey (2D). The determination was achieved by diazotization enzymes in milk and dairy products. Paggi et. al. (23E) de-
and coupling with 3-hydroxy-2,7-naphthalenedisulfonate (R- termined protease activity in milk samples by incubation with
salt) followed by reverse-phase HPLC. Scattering problems NaN3 followed by the addition of TCA, filtration, and
encountered in surface reflectance measurements of meat color treatment with fluorescamine for fluorometric detection.
were resolved by Iversen and Palm (3D) by using an inte- Fukal et al. (12E) reported the residual activity of partially
grating sphere and converting reflectance spectra to the C.I.E. inactivated papain is markedly dependent on the substrate
1976 L*a*b* coordinate color system. and increased as the relative molecular weight of four tested
substrates decreased. The residual pepsin and chymosin
ENZYMES activity in cereal was investigated by Majeed and Ernstrom
General books on enzymes include Plant Proteolytic En- (19E) using different extraction procedures and assay by a
zymes, edited by Dalling (7E), and Methods of Enzymatic linear agar diffusion method. Herrmann and Krause (15E)
Analysis, edited by Bergmeyer et al. (IE), which cover described the apparatus and method to determine the activity
methods for peptidases, proteinases, and their inhibitors. of rennet preparations by measuring change in viscosity of
a solution containing casein. An assay technique for quan-
a-Amylase and /3-amylase of Tritical flour were separated by
affinity and hydrophobic chromatography and assayed tifying milk-clotting enzymes in milk and whey solutions
spectromphotometrically with p-nitrophenylmaltoheptoside without prior removal of casein was described by Carlson et
substrate by Carlsson et al. (4E). Finney (10E) modified the al. (3E). Yada and Nakai (30E) investigated the relation
falling number test to quantitate a-amylase in wheat flour. between physiochemical properties and enzymic activity
A procedure was developed for the determination of fungal (milk-clotting to proteolysis activity ratio) of aspartic pro-
a-amylase additives in wheat flour by Van den Noortgaete teinases by using the mltivariate statistical technique known
as principal component analysis. A method for the quanti-
(28E) using the viscosity attained in a Brabender amylograph
as a measure of amylase activity. Taeufel et al. (27E) found tative determination of trypsin inhibitor in vegetables and
the S-tablet test (Sofa test) to be simple, rapid, and accurate meat products was described by DePalozzo and Callejas (8E),
for determining a-amylase in ceral products. Cinco et al. (6E) who suggested the possibility of applying it for the deter-
described a procedure for the isolation and determination of mination of soybean added to food.
a-amylase inhibitors in white and red kidney beans and de-
termined their heat stability. A rapid procedure and the FATS, OILS, AND FATTY ACIDS
equipment required for the assessment of preharvest sprout Analysis of Fats and Oils, a book edited by Hamilton et
damage of cereal grains was described by Jensen et al. (16E) al. (65F), is recommended to the fats and oils analyst for review
who proposed the method as a basis of a grading system that and reference. Several review articles have been also pub-
reflects the distribution of enzyme activity among individual lished. Arens et al. (7F) published a survey where methods
grains. are described in detail for determining soap and phosphatide
Demmer and Werkmeister (9E) differentiated fresh and residues in refined oils. Christie et al. (24F) published a review
thawed pork by determining the presence of the enzyme /3- on recent developments in lipid analysis, and a review on rapid
hydroxyacyl CoA dehydrogenase (NADH). Two methods for methods for the quality control of edible oils was published
distinguishing between fresh and frozen thawed liver were by Frega et al. (47F).
described by Gottesmann and Hamm (13E)\ in one method Goh et al. (56F) reviewed the minor constituents of palm
the released HADH activity is measured photometrically by oil, and a review and discussion on simple chemical and
means of an enzyme test and in the other by means of a simple physical methods for measuring flavor quality of fats and oils
color reaction. A spectrofluorometric procedure using 4- was published by Gray (60F). Guan et al. (62F) presented

224 R * ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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a review on effects and analysis of secondary lipid oxidation The method has been adopted offical first action by AOAC.
products, and Homberg (72F) authored an article on studies Hibino et al. (68F) reported on the analysis of fatty acids
of sterols in foods, discussing the isolation of lipids and sterols in concentrated polyunsaturated fatty acid fish oil by using
from foods, the use of sterol determination in food research, diethylene glycol succinate (DEGS) packed column. Overlaps
and the sterol content of some plant and animal fat. of Me ester polyunsaturated fatty acid peaks when using
A review on nondestructive monitoring, application of DEGS column were resolved by GC-MS and by GC on a
techniques as IR and NMR spectroscopy in monitoring meat Carbowax 20M capillary column. Hibino et al. (69F) reported
and meat products, discussing lipid and protein determination the results of a collaborative study for the determination of
in meat by NMR, and lipid, protein, and moisture determi- eicosapentaenoic (IPA) and docosahexaenoic (DHA) in
nation by IR spectroscopy, was published by Kopp et al. (94F). polyunsaturated fractionated fish oil.
TLC and GC procedures for separation and characterization Homer (73F) described methodologies for the capillary gas
of unsaponifiable matter in vegetable oils were published by chromatography of milk fat and fatty acids and compared
Lercker et al. (103F). Luf (108F) published a review on (74F) GC methods for the determination of fatty acid methyl
analysis of fatty acids. Monacelli et al. (126F) reviewed the esters and sterols in milk fat and modified milk fat with other
advances in fat analysis with emphasis on GC, NMR, and methodologies. Gas chromatographic methods yielded valid
HPLC. Organoleptic analysis of flavors by physicochemical estimates of total polyunsaturated fatty acids in milk fat
methods was the subject of a review by Mordret et al. (129F). compared with lipoxidase methods and trans fatty acid value
Reineccius (149F) published on isolation, separation, and in milk and butter compared well with data obtained by IR
characterization of flavor compounds in lipids. Schiefer et spectrometry. Kawai et al. (89F) determined the fatty acid
al. (151F) reviewed the enzymic-spectrophotometric methods composition of butterfat, margarine (containing milk fat), and
of determination of phosphatidylcholines in biological ma- coconut oil by GC of their methyl esters; short-chain (C4-C13
terials, e.g., amnistic fluids and blood, and of lecithins in and C16;0) fatty acid methyl esters were measured by tem-
foodstuffs. Schwarz (158F) published a review on methods perature-programmed GLC on a 15% OV-275 column and
for determining fatty acids and glycerides in foods, the physical long chain (C14-C24) by isothermal GLC on a column of di-
properties of fats and their sterol content, and the determi- ethylene glycol succinate. A comparison of two standard
nation of the degree of fat oxidation. The methodologies used titrimetric procedures for determination of free fatty acids
to determine geometric (cis-trans) and positional isomers of content in oils extracted from oilseeds and vegetable oils was
unsaturated fatty acids in fats and oils were reviewed by done by Kershaw (90F). Kroll et al. (98F) reported on the
Strocchi (117F). results of a collaborative study on a FAME-GC method for
Totani (183F) reviewed the microquantitation methods for determining combined and free monomeric fatty acids in
determination of lipid peroxides in foods, especially vegetable refined soybean oil, used frying fats, and two oil mixtures.
oils. Methods including TBA test, cyclooxygenase procedure, trans-Octadecenoate and trans-9-trans-12-octadecadienoate
glutathione peroxidase method, chemiluminescence, GC, were analyzed by Lin et al. (105F) in fresh lean and fatty
HPLC, and TLC-FID methods for determining lipid per- tissues of pork and beef by argentation TLC and capillary GC.
oxides were discussed. Usuki (186F) published a review on Magak’yan et al. (110F) determined chanakh cheese fatty
relations between chemiluminescence and peroxide, acid, acids by GC. Plamitic, myristic, stearic, and lauric acids were
carbonyl, and iodine numbers in thermally oxidized and au- the predominant saturated fatty acids and oleic, linoleic, li-
tooxidized edible oils. Ward (193F) presented a review on the nolenic, and dodecadienoic acids were the predominant un-
use and limitation of TBA assay in estimating lipid oxidation saturated acids.
in food. Yunusova et al. (204F) published a review on methods Mallet et al. (111F) studied derivatization techniques for
of stereospecific analysis of foods for determining the mo- improving the separation of fatty acid esters by capillary GC,
lecular structure of glycerides and phospholipids. Spectro- by modifying the carbon chain properties by addition reactions
photometric methods for analysis for glycerides, fatty acids, on the double bonds.
and their derivatives by use of carbon-13 NMR were reviewed Matsui et al. (120F) identified fatty acids in human blood
by Zimniak (206F). serum and in soybean and colza oils by determining the
Athnasios et al. (9F) published a method on the determi- equivalent chain length (ECL) values of C14_22 fatty acid esters
nation of Ms,m-methylene interrupted polyunsaturated fatty using various glass capillary columns. Misir et al. (124F)
acids in fats and oils by capillary GC on a 60M SP 2340 proposed and evaluated a rapid transesterification method
capillary column. Gas chromatographic results agreed well for the preparation of fatty acid methyl esters (FAME) for
with those obtained by an enzymic lipoxygenase method. gas chromatographic analysis using tetramethylammonium
Bannon et al. (10F) determined the composition of fats with hydroxide in methanol and compared it with conventional
fatty acids containing four or more carbon atoms by gas estrification methods for a variety of vegetable and animal
chromatography of fatty acid methyl esters (FAME); the oils and fats. Better recoveries of low chain fatty acids (e.g.,
optimum method of methylation was discussed. C4) and also for unsaturated C13 fatty acid isomers where
Aldolf et al. (2F) authored a paper on the isolation of co- achieved when the rapid method was employed.
3-polyunsaturated fatty acids and methyl ester fish oils by Moneam et al. (127F) analyzed total fatty acids extracted
silver resin chromatography. from S. terebenthifolius berries with light petroleum, diethyl
Boniforti et al. (17F) determined positional and geometric ether, or CHCl3:MeOH (2:1) by FAME-GC-FID. Pyroly-
isomers of unsaturated fatty acids commonly found in vege- sis-GC analysis was used by Nazer et al. (136F) as an iden-
table oils by converting the methyl esters of fatty acids to tification method of fats and oils. Pyysalo et al. (146F) re-
epoxides with m-chlorobenzoic acid, followed by capillary ported capillary GC methods coupled with detection by mass
GC-MS. Cao et al. (20F) determined palmitic, oleic, linoleic, fragmentation and retention index monitoring (RIM) for fat
linolenic, arachidonic, and erucic acids in rapeseed by GC-FID analysis. A rapid method for the determination of double
of their methyl esters. bond positions in monounsaturated fatty acids by GC-MS
Conway et al. (27F) used hydrazine to hydrogenate the was reported by Shibahara et al. (162F). Strocchi et al. (176F)
double bond of cyclopropenoic fatty acids of seed oils without determined unsaturation in hydrogenated soybean and palm
cleaving the cyclic system and with minimum effect on the oils and in margarine oil by GC on a 15% OV-275 on 100-120
degree of unsaturation of the remaining double bonds prior mesh Chromosorb P AW/DMCS at 220° and nitrogen as
to GC analysis of methyl esters. Gas chromatography was used carrier gas.
to analyze the fatty acids of hazelnut oil as reported by De- Stack et al. (174F) developed a capillary GC-FID method
mirbas (36F). Dupuy et al. (37F) used direct sampling ca- for the simultaneous quantification of fatty acids (as methyl
pillary gas chromatography for determining volatiles and esters) and sterols (silyl esters) in orange juice. Spangelo et
flavors of vegetable oils. Gere et al. (54F) determined cyclic al. (173F) determined individual free fatty acids in milk by
fatty acids formed during heating of sunflower and rape oils GC-FID using a column of 10% SP-2330 on Chromosorb W.
by GC-FID of their methyl esters on Carbowax 20M with Sumimoto et al. (179F) described a capillary GC-FID method
hydrogen as carrier gas. Gildenberg et al. (55F) reported the for determination of fatty acids in food preparations including
results of an international collaborative study of a gas chro- fish, meat, and eggs, oils and fats, and beans, milks, grains,
matographic method for determination of trans unsaturation and confectionary products. With the exception of beans, C14;0,
in margarine. Coefficients of variation were consistently better Ci6:o> C12;0, Ci3:i, C13:2, and C18;3 were detected in all food
for the determination of total trans acids by GC vs. IR analysis. preparations. In addition C20:4, C20;6, and C22;6 were detected

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 225 R
FOOD

in fish but were absent or present only in trace levels in other system were used. Samples were applied to chromatorods and
food preparations. organic components were pyrolyzed in hydrogen flame, re-
Takagi et al. (18IF) analyzed soft margarines for their fatty corded, and calculated automatically, as reported by Pisareva
acid content using capillary GC of fatty acid methyl esters. et al. (142F). Ratnayake et al. (147F) also used Iatroscan-
The content of the geometrical and positional isomers of oleic thin-layer chromatography-flame ionization detection for the
acid was determined by GC of the ozonolysis products of the rapid analysis of canola gum lipid composition. Sebedio et
cis and trans monoenoate fractions, separated by argentation al. (159F) quantitatively analyzed the methyl esters of fatty
silicic acid TLC. Tsuda et al. (185F) described a method for acids geometrical isomers and of trilgycerides differing in
GC determination of sorbitan fatty acid esters in confectionary degree of unsaturation by Iatrascan TLC/FID using silver
products as sorbitan monostearate. Erucic acid content in nitrate impregnated rods.
rape oil was determined by Wang et al. (191F) by FAME- Fraser et al. (44F) detected chlorophyll derivatives in
GC-FID on columns of QF-1, OV-17, or SE-30. Yamazaki et soybean oil using reversed-phase HPLC. Antioxidants in
al. (201F) determined erucic acid in rapeseed by near-IR tallow and vegetable oils were determined by Indyk et al. (79F)
reflectance spectroscopy. by normal-phase HPLC with UV detection at 280 nm. Wei
Woo et al. (196F) described modified techniques for im- et al. (194F) determined BHA, BHT, and PG in oil fat and
proved GC quantification of free fatty acids in dairy products. food by TLC and GC.
Zhou et al. (205F) analyzed free fatty acids of several vegetable Maruyama et al. (117F) separated and quantitated un-
oils by capillary GC of fatty acid methyl esters on a column derivatized saturated and unsaturated monoacylglycerols by
coated with cross-linked methyl silicone. IUPAC commission reversed-phase HPLC and UV detection at 210-215 nm.
on oils, fats, and derivatives (83F) published the results of two Ascorbyl palmitate was determined in vegetable oil and lard
collaborative studies for the determination of butyric acid in by reversed-phase HPLC and UV detection by Vicente et al.
butter fat alone or blended with other fats. Recseg et al. (188F).
(148F) determined fatty acid trans isomers in margarine and Cholesterol was determined by Arai et al. (5F) using an
hydrogenated rape oil by GC on packed columns of OV-275 integral multilayered analysis element. Hurst et al. (77F)
or Silar IOC. The results agreed well with those by a standard presented an HPLC method for the determination of chole-
IR spectrometric method. sterol content as an indicator of egg solids. Kou et al. (96F)
HPLC preparative separation of milk fatty acids as their described a method for the determination of 25-hydroxy-
methyl derivatives was reported by Christie et al. (25F) with cholesterol in a wide range of materials including foods. Lipid
satisfactory resolution of C4-C18 fatty acids esters using Li- extracts were initially purified on octadecyl silicic acid car-
Chrosorb 10 RP column with isocratic elution with MeCN:H20 tridges and by reversed-phase HPLC before quantitation by
(95:5) as mobile phase. Hirata et al. (70F) reported on the normal-phase HPLC on a silicic acid column and a variable
analysis of free saturated, unsaturated, and branched fatty wavelength detector. 25-Hydroxycholesterol was not detected
acids by reversed-phase HPLC with MeOH containing 1 % in lard, cream, fresh egg yolk, or spray-dried egg yolk powder
phosphoric acid as mobile phase and low UV detection. but was detected and its identity confirmed by mass spec-
Hwang et al. (78F) used HPLC for quality evaluation of trometry in egg yolk powder after heating at 110° for 4 days.
sesame oil. Kihara et al. (9IF) described a simple precolumn Maerker et al. (109F) determined a number of standard
saponification and esterification reactor for preparation of cholesterol oxides by direct on-column GC (5% phenyl-
phenacyl, p-bromophenacyl, or 2-naphthyl esters of fat and silicone-coated capillary column with temperature program-
oil fatty acids prior to HPLC analysis. Lu et al. (107F) de- ming) with good resolution, recommending its use to measure
termined fatty acid composition in rapeseed oil during hy- these oxides in foods.
drogenation by HPLC on Nucleosil C18,5 ^m, and MeCN as Fischer et al. (40F) determined oxycholesterols in various
mobile phase. Matsumoto et al. (121F) reported a micro liquid foods by GC and in foods of animal origin by GC-MS (41F).
chromatographic system directly coupled to a quadrupole mass Masoom et al. (118F) described a method for determination
spectrometer through a vacuum nebulizing interface that was of cholesterol by flow injection analysis with immobilized
applied to the analysis of free fatty acids of bean oil, rape oil, cholesterol oxidase and hydrogen peroxide produced was
palm oil, and milk fat. detected amperometrically. Masson et al. (119F) determined
Netting (137F) presented a one-pot procedure for the sa- cholesterol in butter by rapid saponification with KOH and
ponification of fats and oils, and the subsequent estrification elution of the unsaponifiable fraction from a Celite 545 column
of the resulting fatty acid salts with pentafluorobenzyl bromide with chloroform followed by silylation and GC on a column
followed by the separation of the pentafluorobenzyl esters of 3% SP-2250 on Supelcoport (100-120 mesh) at 245° with
primarily on the degree of unsaturation by normal-phase FID.
HPLC. Ritchie et al. (151F) used normal-phase HPLC to Mordret et al. (128F) developed a GC method for the de-
separate lipid classes as was exemplified by separation of termination of cholesterol in fats and oils and in margarine
methyl ricinoleate, ricenoleic acid, and ricinoleyl alcohol and using cholesterol as internal standard. C-7 oxidized cholesterol
fractionation of cocoa butter and superglycerinated partially derivatives (C-7 OCDs) in muscle and other foods were de-
hardened rape seed oil. termined by Park et al. (138F) without saponification by
Robinson et al. (152F) compared RI, UV, and mass de- chromatographing the total lipid extracts on silica gel to
tectors for HPLC analysis of glyceride mixtures. A new double concentrate trace sterol oxides from triglycerides, cholesterol,
column HPLC method for rapid separation of fatty acids and phospholipids. Sterol oxides in eluates were determined
(C12-C22) was published by Sato (156F). Vioque et al. (189F) by normal-phase HPLC on a /uPorasil column with 7% 2-
separated free fatty acids by RP-HPLC after conversion to propanol in hexane, detecting 7-ketosterols at 233 nm and
p-phenylazophenacyl esters. An HPLC method was developed 7-a-hydroxy cholesterol at 208 nm. Recoveries of C-7 OCDs
by Wood (197F) for the analysis of seed oils containing cy- added to beef approached 100%. Pancake mix, french fries,
clopropene fatty acids after hydrolysis and conversion of the raw beef, fried chicken, cooked hamburger, beef jerky, and
acids to phenacyl derivatives using a two-column system that liver sausage were analyzed. Sugino et al. (178F) developed
allowed the resolution and quantitation of malvalic, sterculic, a method for the determination of cholesterol epoxides as the
and dihydrosterculic acids as well as other fatty acids in p-nitrobenzoate derivatives in spray-dried egg products using
cottonseed and sterculia foetiola oils. Yabe et al. (198F) reversed-phase HPLC.
determined gossypol in edible cottonseed oil by HPLC on a Bianchini et al. (13F) reported on the determination of
Hypersil M05 column with MeCN-N20-THF (80:18:2), pH sterol and triterpene alcohol acetates in natural products by
2.5, as mobile phase and UV detection in 235 nm. reversed-phase HPLC and GC-MS. Brumley et al. (19F)
Lipid fractions (phospholipids, sterols, glycerides, and free identified sterols in vegetable oils by capillary GC-MS-EI of
fatty acids) were determined in lipids from grill and frozen sterol butyrate esters and determined them by packed column
fish by a method combining separation with a hexane-chlo- GC-FID. Chaouch et al. (21F) determined tocopherols in
roform-ether-potassium acetate (85:15:17:1) as a mobile phase peanut, corn, olive, soybean, and sunflower oils by HPLC and
and flame ionization detection (FID). Analyses were per- by differential pulsed voltammetry.
formed on an Iatroscan analyzer using silica gel coated Fujimoto et al. (48F) determined sterols in edible oils by
chromatorods. For separation of saturated and unsaturated reversed-phase HPLC and UV detection at 205 nm. IUPAC
triglycerides, chromatorods impregnated with silver nitrate commission on oils, fats, and derivatives (81F, 82F) published
and mobile phase of CHCl3-benzene-acetic acid (10:90:1) two papers on the determination of total sterol in fats and

226 R ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
 FOOD

oils. Gracian Tous et al. (59F) described a method developed tributyltin in marine food products. Fish or shellfish was
by collaborative study for the determination of erythrodiol homogenized with HCl-MeOH to convert tributyltin oxide
in vegetable oils and the separation of eight structurally closely to tribyltin chloride which after fractionation using a silica
related sterols on C8 and C18 reversed-phase columns with UV gel column was derivatized to its trimethylsilyl derivative and
detection at 206 nm was reported by Holen (71F). analyzed by GC.
Hortsmann et al. (75F) described a rapid method for iso- Direct inlet system/ammonia chemical ionization mass
lation of sterols from plant and animal fat and oils and for spectrometry was used by Murata et al. (132F) for analysis
determining the content of these fats and oils in foods. Jonker for triglycerides of linseed, safflower, and lettuce seed oils.
et al. (85F) reported an improved method for determination A method for the quantitative determination of the tri-
of free, esterified, and glycosilated plant sterols in foods. A glyceride species composition of vegetable oils by reversed-
method for the detection of sterol epoxides in foods by col- phase (RP) HPLC via flame ionization detection was described
orimetric reaction with picric acid was reported by Lee et al. by Phillips et al. (141F), separations were achieved by using
(101F). Mrugasiewicz et al. (131F) analyzed sterols in corn Zorbax ODS columns with gradient elution with methylene
by extraction with chloroform and determination at 615 nm chloride in acetonitrile. The method was applied to butter
after reacting with the Liebermann-Burchard reagent. Na- before and after randomization, soy oil, and pure olive oil.
sirullah et al. (134F) reported a TLC method for the analysis Pocklington et al. (143F) reported on the development, by
of unsaponifiable matter of edible and nonedible seed oils. collaborative study, of a standardized method for determi-
Naudet et al. (135F) reported on a new official procedure nation of the component triglycerides of animal and vegetable
(IUPAC 2.404) for sterol determination in plant or animal oils. The procedure involves the separation of triglyceride
lipids. groups containing the same number of C atoms, by direct GC
Separation of triglycerides and hydrocarbons from seed oils of the lipid solutions on packed columns under tempera-
by HPLC with IR detection was reported by Bhati et al. (12F). ture-programmed conditions. Podlaha et al. (144F) studied
Triglycerides of C^-C^ of cocoa butters from different parts the triglyceride (TG) composition of 28 cocoa butter samples
of the world were separated by GC by Chaveron et al. (23F). by HPLC.
Christopoulou et al. (26F) used a high-performance size ex- Shukla et al. (163F) separated triglycerides from cocoa
clusion chromatographic method for the separation and butter by HPLC on two Spherosorb reversed-phase columns
quantitation of fatty acids and mono-, di-, and triglyceride with MeCN-THF (73:27) as mobile phase and UV monitoring
mixtures. Separation was achieved on two columns packed at 220 nm. The analysis of triglycerides in oils and fats by
with 5-/um styrene/divinylbenzene copolymer and connected liquid chromatography with laser-light scattering detector was
in series, using toluene as eluent, and components were reported by Stolyhwo et al. (175F). Toeregaard et al. (182F)
monitored by refractometry. Fiebig (38F) reported re- separated palm oil, milk fat, and model mixtures of intact
versed-phase HPLC method for triglyceride separation on glycerides by capillary GC with OV-101 as stationary phase
LiChrosorb RP18 with propionitrile as mobile phase. Frede on the basis of their number of C atoms and on the number
(46F) described factors for improving triglyceride separation of unsaturated fatty acid in the triglyceride molecule.
by reversed-phase HPLC using propionitrile as mobile phase Wada et al. (190F) reported on triglyceride analyses of
and RI detector. vegetable oils and animal fat by GLC, field-desorption mass
Geerarert et al. (50F) discussed several capillary GC and spectrometry (FD-MS), and HPLC. The combination of these
HPLC methods fro triglyceride analysis including separations techniques provided significant information by which the
by carbon number, by number of unsaturated fatty acids major molecular species of triglycerides in naturally occurring
(within each Carbon no.), and by double bond locations fats and oils could be estimated. Yoshida et al. (203F) reported
(ozonolysis), using reversed-phase HPLC, and presented a a simple and rapid method for the enzymic hydrolysis of
scheme for the analysis of chocolate fats (including cocoa triacylglycerols based on the enzyme deacylation of triacyl-
butter, butter oil, cocoa butter equivalents, and vegetable oils) glycerols on a single TLC plate followed by analysis of the
and for the analysis of compound chocolate. Geerarert et al. hydrolytic products separated from the reaction zone by
(51F, 52F) reported on the capillary GC-FID analysis of multiple development of the plate. The method is applicable
triglycerides of palm, grape seed, cottonseed, soybean, olive, to the analysis of most natural fats and oils.
corn, peanut, rape, coffee, and coconut oils and of butterfat, A semimicro method for the determination of iodine values
egg yok, and chicken fat. The triglycerides were separated of lipids by bromide ion selective electrode was developed by
according to chain length, and the retention was highest with Abdel-Moety et al. (IF). Kolarovic et al. (93F) reported an
triglycerides containing the most unsaturated acids. on-line gas chromatographic method which allows direct
A paper on HPLC of triglyceride determination with gra- calculation of iodine values of edible fats and oils. Sheeley
dient elution and mass detection was published by Herslof et al. (161F) compared the determination of iodine values in
et al. (67F). commercial soy lecithin samples and lecithin standards by the
Kamat (86F) reported a method for the determination of AOCS official method (Wijs) and by measurement of proton
mono-, di-, and triunsaturated triglyceride content of corn, NMR signals for vinylic protons and found good agreement
soybean, and rape oils by TLC-FID. In another paper, Kamat between the two methods.
(87F) used a TLC-FID method to quantitate dipadmitoolein, Agienko et al. (3F) reported a highly sensitive thiobarbituric
palmitodiolein, and triolein by quantitatively oxidizing them acid (TBA) test for predicting the resistance of fats in milk
with KMn04 to the corresponding azelagolycerides which were substitutes to oxidation during storage. Pokorny et al. (145F)
separated and determined by TLC-FID using squalene as an described a modified method for determination of 2-thio-
internal standard. barbituric acid value in fats and oils, which utilizes 1-butanol
Lee (102F) developed two methods for the separation of as the sole solvent. The method was satisfactory for evaluating
medium chain triglycerides using reversed-phase HPLC. Both lard, cooking fat, soybean, sunflower, and rapeseed oils in stage
methods employed C18 HPLC column and isocratic elution, of beginning rancidity. A simplified plate diffusion technique
with a differential refractometer detector, and a UV detector for the determination of lipid antioxidant activity was de-
at 210 nm using Trimonanoin as the internal standard for scribed by Araujo et al. (6F).
quantitation. The method is suitable for milk, whey, and Asano et al. (8F) described a method for the detection of
sobyean based matrices, and with minor modification, medium lipid peroxides in foods. Inverse gas chromatography was
chain triglyceride levels ranging from 10 to 50% of total fat found by Bird et al. (14F) to be suitable for study of the
could be determined. oxidation of vegetable oils. Blumenthal et al. (15F) developed
The determination of the structure of glycerides in olive a method for determination of oxidized (polar) substances in
oil was the subject of a publication by Mariani et al. (112F). fat and a new quick test for the detection and semiquantitation
Marjanovic et al. (113F) determined triglycerides (C2g-C52) of alkaline contaminant material such as soap in fresh and
in fats (cocoa butter and its substitutes and lard) by GC on used vegetable frying oils and animal/vegetable oil blends
short capillary SCOT (support-coated open tubular) columns (16F).
and in milk fat by high-temperature GC on a 1 % Dexsil 3-SS Bradley (18F) reported the results of a collaborative study
column (114F). Martinez-Castro et al. (115F) used tetra- on methods of determining milk fat. Chaveron et al. (22F)
methylammonium hydroxide as catalyst in the methylation reported the results of a collaborative study on a GC method
of both free fatty acids and glycerides of fat prior to analysis for determining vegetable fats in chocolates on the basis of
of milk fat by capillary GC. Morisaki et al. (130F) determined variations in the triglyceride composition. Low concentrations

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 227 R
FOOD

of milk fat were determined by Cotton (28F) in skim milk and nological treatments or frying by FAME-GC.
cream plant wastewaters using a modified procedure of The sensory properties of volatile lipid oxidation product
Roese-Gottlieb with preconcentration of the sample. and methods for the determination of lipid oxidation in food
A comparative study of six analytical methods for quality were reviewed by Hall (63F). Hall et al. (64 F) studied the
evaluation of frying fats was published by Croon et al. (29F, formation of secondary lipid oxidation products in spray-dried
30F). Cruddgington (3IF) reported the results of a study of cream and whole milk powders by static and dynamic
the determination of fat in dried milk. Bengtsson (11F) re- headspace sampling and gas chromatography. Hara et al.
ported that the near-iR reflectance method for determining (66F) used reversed-phase HPLC with RI detector for the
oil content in rapeseeds was superior to the gravimetric separation and determination of autoxidized ethyl oleate,
analytical methods. Davies et al. (32F) derived equations for linoleate, and linolenate. The hydroperoxides were separated
estimating oil concentration in mayonnaise containing 11-75% according to the number of double bonds and hydroperoxy
oil in mayonnaise-based salads using near-IR. Fat content groups and geometrical (cis-trans and trans-trans) configu-
in meats (ground pork, beef, and chicken) was determined by ration of double bonds, but not on the basis of position of
near-IR reflectance as reported by Nagao et al. (133F). De hydroperoxy groups.
Koning et al. (33F) reported the results of a critical study on Kikugawa et al. (92F) reported on the chromogenic de-
a number of lipid extraction methods from fish meal. termination of lipid hydroperoxides by reacting with sesamol
The application of proton NMR in food analysis was dis- dimer, in the presence of Hb. The violet-colored reaction
cussed by Defour (34F). Specific examples considered were product had an absorption maximum at 550 nm. By this
the determination of the solid/liquid phase ratio in refined method 0.1 jumol of methyl hydroperoxide could be detected
and hydrogenated coconut oil and the determination of total with higher sensitivity than by the conventional peroxide value
fat in chocolate. Gambhir et al. (49F) simultaneously de- (POV) method. Lang et al. (100F) described a sensitive
termined the moisture and oil content in rapeseed-mustard method for the determination of the lipid peroxidation product
by pulsed NMR. Guan et al. (62F) determined fatty acids 4-hydroxynonenal in biological samples as well as lipid-con-
in vegetable oil by carbon-13 NMR. Lambelet et al. (99F) used taining foodstuffs. Mikula et al. (123F) reported on reaction
low-resolution pulsed NMR to determine solid content in conditions for measuring oxidative stability of oils by ther-
cocoa butter and edible oils. mogravimetric analysis. Headspace chromatographic analysis
A rapid NMR method for fat analysis (total and solid fat of heated soybean oil was applied by Snyder et al. (168F) to
content) was developed by Leung et al. (104F). Patersson et determine the effect of hydrogenation and additives on the
al. (140F, 139F) reported a general pretreatment and meas- formation of total and individual volatile components.
uring procedure for solid fat content (SFC) determination Sotirhos et al. (171F) used HPLC for the analysis of oxi-
using pulsed NMR on tempering fats and tempering fats in dative and polymerized decomposition products in commercial
blends with milk fat. Low-resolution NMR was used by Renou vegetable oils and heated fats. Szumilak et al. (180F) reported
et al. (150F) for determining fat content in meat products. that precision of the iodometric method for determining
Ryseva et al. (155F) determined the level of solid triglycerides peroxides in soybean was markedly improved by using po-
in cocoa butter and in some of its commercial substitutes. tentiometric measurements. Valentova et al. (187F) deter-
Wang et al. (192F) presented a new double-tube addition mined total polar oxidized fatty acids of the fat fraction iso-
method for measuring the oil content of seeds and other food lated from dehydrated meat and dehydrated chicken soup and
samples by pulsed NMR with diode detection. of fat from frying doughnuts and from roasted peanuts by
Phospholipid separations from vegetable oils were studied reversed-phase HPLC with RI detector.
by Ajana et al. (4F) by dialysis of crude oils against petroleum Woestenburg et al. (195F) reported the results of an in-
ether which allowed the recovery of almost all phospholipids terlaboratory test for examining the performance of the au-
(98%), fractionation by HPLC, and TLC. Fozy et al. (42F) tomated Rancimat method for determining the oxidative
described a rapid enzymic spectrophotometric method for stability of edible oils. Yagi et al. (199F) used a methylene
determining lecithin in cocoa, chocolate meases, and soybean blue derivative for the determination of lipid peroxides in
phosphatides. Goh et al. (57F) determined total phospholipids foods. Results obtained by this method coincided well with
in crude palm oil using colorimetry. Analysis of phospholipids those by the iodometric method and paralleled those obtained
in soy lecithin by HPLC was reported by Hurst et al. (76F) by the thiobarbituric acid method. Yamamoto et al. (200F)
who analyzed chloroform solutions of soy lecithin by HPLC detected low level lipid hydroperoxides by chemiluminescence.
using a normal phase (/t-Porasil) column and a mobile phase Deman et al. (35F) developed a new instrument for thermo-
MeCN-MeOH-85% H3P04 (780:10:9) with UV detection at penetrometry of fats, the use of which was demonstrated with
205 nm. butter and margarine. Fletcher et al. (41F) published a report
Ishiguro et al. (80F) used different methods to determine comparing the various procedures for determining total yolk
lecithin in food preparations including precipitation with lipid content. Frede et al. (45F) studied the variation in
acetone, colorimetric measurement of phosphorus, TLC, and melting and crystallization temperatures of milk fat by dif-
GC. Acetone precipitation gave high results because of the ferential scanning calorimetry (DSC) and discussed the use
presence of coprecipitated sterols and other substances while of DSC for detecting foreign fats in milk fat. Gegiou et al.
the colorimetric method gave variable results. TLC allowed (53F) reported on methodology for determining cocoa butter
the determination of total lecithin as well as the individual equivalents (CBE) in plain and milk chocolate. Grover et al.
phosphatides and GC was satisfactory for soybean lecithin. (6IF) detected palas oil in ground nut and vegetable oils by
Martovshchuk et al. (116F) described an apparatus for analyzing the unsaponifiable fraction of fat by TLC with
measuring the interphase tension between a vegetable oil and fluorescence detection under UV irradiation.
water. The value obtained is a measure of the hydrophilic Methods for determining glycerides and free fatty acids in
phospholipid content of the oil and thus of its degree of re- butter flavoring materials were examined by Kato et al. (88F).
fining. A method for determination of lecithin in foods by GC was most suitable for free fatty acids and gel chroma-
TLC separation followed by spectrophotometric measurement tography was most suitable for glycerides. Korynova et al.
of phosphorus was developed by Senelt et al. (160F). Sotirhos (95F) determined the butterfat content in chocolate products
et al. (1 70F) reported an HPLC method for soybean phos- by determining the level of butyric acid titrimetrically.
pholipid analysis using a silica column, a mobile phase of Krishnamurthy et al. (97F) detected tricresyl phosphate (TCP)
hexane-2-propanol-water, and UV detection at 210 nm. The and tri-o-crysyl phosphate (TOCP) in contaminated edible
same authors (172F) used both normal-phase and reversed- oils by TLC. TOCP was analyzed by GC-FID for confirma-
phase HPLC for further analytical studies of soybean phos- tion and quantitation of its level in oils. Linfield et al. (106F)
pholipids. Traitler et al. (184F) described the comparative reported a new method for the assay of lipase activity.
determination of phosphatidylcholine (lecithin) species in food Maxwell et al. (122F) described a method for the quantitative
products by fast atom bombardment computer averaging extraction and simultaneous class separation of milk lipids.
integration spectrometry (FAB-CAI). Lipid classes were separated into neutral and polar fractions
Autoxidized fats were studied by Frankel et al. (43F) to by column chromatography on a Celite column with sequential
elucidate the genesis of volatile lipid oxidation products. elution using solvents of increasing polarities.
Thermal homolytic and acid heterolytic decomposition pro- The extraction of lipids and their quantitative classification
cesses were compared using GC/MS. Gomes et al. (58F) in meat products were carried out through the combined use
evaluated thermal oxidation in vegetable oils following tech- of TLC and GC as described in a publication by Miteva

228 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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(125F). Rosenthal et al. (153F) proposed an acid digestion Discriminant and cluster analyses of GC profiles of Wor-
method for fat determination in vegetable foods. Rossell et cestershire sauce volatiles were utilized by Aishima (3G) to
al. (154F) analyzed palm and coconut oil samples for fatty acid, differentiate sauces. The discriminant functions consisted
triglyceride, sterol, and tocopherol composition as well as their of 12/85 peaks which were identified as terpenoids, phenols,
melting properties. Slover et al. (164F) reported on the de- and heterocyclic compounds. Carpenter et al. (11G) related
velopment and use of quality control samples in food lipid GC profiles of orange juice to processing conditions by ap-
analysis. plication of multivariate methods. Izquierdo et al. (32G)
Smidovnik et al. (165F) developed a method for extraction applied principle component analysis to orange juice pro-
and determination of lipids from corn starch, and analyses cessing; the first principle component related to time and
of honey lipids were reported by Smiljanic (166F). Smurygina temperature of storage, and the second principle component
et al. (167F) reported on the results of fat determination in related to cold-filled juices. Four GC peaks of orange juice
rennett and process cheeses by the Gerber and Van Gulik essence, terpinen-4-ol, trans-2-hexenal, an unidentified ses-
methods as compared with results of solvent extraction/gra- quiterpene, and an unknown compound, were used by May-
vimetric method and concluded that the Van Gulik method field et al. (47G) to identify orange varieties.
was superior to the Gerber method with respect to accuracy Chang and Reineccius (13G) developed a 40-m continuous
and reproducibility. Soliman et al. (169F) reported on the dialyzing device to isolate flavors from mixtures as a function
fatty acid composition of triglycerides and 2-monoglycerides of sample and solvent flow rates. Tenax GC was used to trap
of butterfat, beef tallow, cottonseed oil, admixtures of beef intact fruit and vegetable aroma volatiles for subsequent GC
tallow with butterfat, and admixtures of cottonseed oil with analysis by DePooter et al. (17G). The artifact backgrounds
butterfat. The determination of emulsifiers in various foods for Tenax GC and Tenax TA were compared by MacLeod and
was reported by Yomota et al. (202F). Ames (46G)\ Tenax TA was found to be cleaner, and a more
efficient preconditioning regimen was proposed. Methods for
FLAVOR the analysis of encapsulated (emulsified) flavors were screened
by Kernik et al, (37G); acetone precipitation, simultaneous
Several reviews relevant to flavor analysis were published steam distillation/solvent extraction, and Fosslet methods
in 1984,1985, and 1986. Busch (10G) reviewed applications yielded uniform recoveries for both low and high boiling
of tandem mass spectrometry to fruit and spice flavor analysis. compounds. Microwave desorption of thiamine decomposition
Chromatographic methods for the determination of phenols products from graphite was described by Reineccius and
and flavonoids in foods and beverages were covered by Hardin Liardom (5.6G). Toulemonde and Beauverd (67G) described
and Stutte (27G). The limits and perspectives of GC-FTIR a device for injection, trapping, and near instantaneous mi-
in flavor analysis were reviewed by Idstein and Schreirer (31G). crowave desorption for application to fruit volatiles.
Liddle and Bossard (43G) developed methods for the detection Adam (2G) described a method for volatile analytes whereby
of /3-asarone, coumarin, hydrocyanic acid, pulegone, safrole, purged and trapped components are desorbed onto an in-
and a- and /3-thujones in foods and beverages. Reineccius and column cold trap (-100 °C) for concentration prior to GC
Anandaraman (57G) contributed a general review of volatile separation. Cryofocusing of headspace volatiles at the head
flavor analysis. Meat flavor volatile composition, analysis, and of a capillary column by Wylie (75G) afforded enhanced
sensory testing were evaluated by Shahidi et al. (60G). Shaw sensitivity and permitted multiple headspace overlays and
and Moshonas (61G) reviewed the use of mass spectrometry splitless injection. Badings et al. (4G, 6G) developed an au-
in the identification of citrus flavor compounds. Food off- tomatic purge and trap system in which efficiencies were
flavor analyses were discussed by Whitfield and Shaw (74G). dependent upon trap composition, purge flow rate, trap tem-
Capillary GLC of vegetable oil quality was achieved by perature, and total purge gas volume. Dirinck et al. (18G)
direct sampling onto a cross-linked 5 % phenylmethyl silicone developed a dynamic headspace sampling apparatus applicable
column by Dupuy et al. (21G). Artifacts were encountered to GC-MS analysis of strawberry and apple volatiles. Gon-
by Liddle and Bossard (44G) when using hydrogen as carrier zalez and Gra (25G) compared headspace concentration by
gas for the analysis of anethole; dihydroanethole was produced closed circuit circulation through Porapak Q, with simulta-
upon sample injection. Nitz (53G) achieved greater compo- neous distillation/ether extraction, and found the headspace
nent resolution using multidimensional GLC in which fractions technique to be faster and equally as reliable as distilla-
from a packed column are further separated by capillary GLC tion /extraction. Kolb (38G) compared equilibrium and dy-
connected either to MS or a sniffing port. Schreier et al. (59G) namic headspace procedures; in general equilibrium tech-
compared capillary GC-MS and capillary GC-FTIR for niques were preferred for splittless injection, cold trapping,
cherimoya fruit volatile analysis; GC-MS showed greater multiple headspace extraction, and calibration by standard
sensitivity. Greater stationary phase thickness (0.45 ^tm) and addition. Borek et al. (9G) modified a laboratory vacuum
higher temperatures (120 °C) were found by Shibamoto (62G) rotary evaporator condensation coil to achieve nitrogen-driven
to yield better GLC Kovats indicies for alcohols and aldehydes. headspace preconcentration of volatiles from fruits and veg-
Normal-phase HPLC of cold pressed lime oil by Chamblee etables. Food volatile concentration methods, including direct
et al. (12G) provided fractions for GC-MS that yielded 23 new headspace, headspace concentration, Nickerson-Likens, and
components. Rouseff (58G) isolated the bitter limonoids from solvent extraction, were compared by Leahy and Reineccius
grapefruit, obacunone, nomilin, and limonin, by preparative (42G); simultaneous distillation/solvent extraction was found
HPLC. Ginger flavor compounds, 6-shogaol, 6-gingerol, 8- to be most efficient at the 50 ppb level. Using a static
gingerol, and 10-gingerol, were isolated by Smith (63G) using headspace technique, Voilley and Bosset (71G) determined
reverse-phase HPLC with electrochemical detection. Van der the partition and activity coefficients for model flavor com-
Greef et al. (69G) applied LC-MS, field desorption MS, and pounds in high viscosity media.
high resolution mass electron impact MS to identify the major Strecker-type compounds in cheddar cheese were isolated
oleoresins of black pepper. and analyzed by Dunn and Lindsay (20G) using Tenax-GC
High-resolution ’*C NMR has been applied to both qual- headspace trapping and acetonitrile extraction; Tenax-GC
itative and quantitative analysis of essential oils and natural trapping was found most suitable for routine testing. Cheese
flavors by Kubeczka and Formacek (40G). Ravid et al. (55G) headspace volatile GC profiles generated by steam distillation
were able to quantitatively resolve (+) and (-) enantiomers were found by Lin and Jeon (45G) to be dependent upon
of natural and synthetic linalool by XH NMR using a chiral cheese age and solids concentration in the cheese slurry. A
lanthanide shift reagent. The enantiomers of 7-lactones were headspace volatile trapping technique for powdered milk and
evaluated using JH NMR by Weinstein et al. (73G) using the dairy products was developed by Badings and DeJong (5G).
chiral solvation reagent, (R)-(-)-2,2,2-trifluoro-l-(9-anthryl)- Horita and Hara (29G) found that more low boiling com-
ethanol. pounds are recovered from tea by using simultaneous steam
George (24G) characterized passion fruit flavors by Car- distillation/ether extraction than by using vacuum stripping;
bowax 20M capillary GC-MS using chemical ionization with distillation time was deemed critical. Hutt and Herrington
OH". Volatile sulfur compounds from garlic oil were identified (30G) identified and quantitated the bitter principle in zuc-
by Vernin et al. (70G) using a variety of GC-MS techniques chini, curcurbitacin E, using both reverse-phase TLC and
including electron impact and positive and negative chemical reverse-phase HPLC. 4-Hydroxynoneal, a lipid peroxidation
ionization and single ion monitoring enhanced component product, was extracted into methylene chloride, cleaned up
resolution. on an ODS extraction column, and determined using re-

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 229 R
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verse-phase HPLC by Lang et al. (41G). A procedure suitable method for the analysis of theaflavin and its mono- and di-
for dairy product QC was developed by Mills (48G) whereby gallate derivatives in tea solution by complexation with 2-
protein volatiles are trapped on a porous polymer, desorbed, aminoethyl diphenyl borate (flavognost) and spectrophoto-
and resolved by capillary GC. Mori and Kiuchi (51G) pre- metric measurement of the complex at 625 nm. An improved
ferred direct solvent extraction over simultaneous distilla- reverse-phase HPLC method for vanillin and related flavor
tion/solvent extraction for quantitative detection of pyrro- compounds in vanilla extract was developed by Guarino and
lidones, pyrroles, pyrazzines, and thiazoles in miso. Allyl Brown (26G). This method is superior to the AOAC method
isothiocyanate, 3-butenenitrile, and acetic acid were identified because ethyl vanillin and p-hydroxybenzaldehyde can be
in mustard paste by Mounie et al. (52G) using solvent ex- resolved.
traction and GC analysis. Nunez and Maarse (54G) preferred
simultaneous steam distillation/ether extraction over static IDENTITY
and dynamic headspace methods and strip/trap methods for Chaveron et al. (12H) determined the presence of foreign
the analysis of grapefruit volatiles. Ovine fat volatiles were plant fats in chocolate by GLC analysis of the C^-C^ tri-
profiled by Suzuki and Bailey (65G) by heating the oil, purging glycerides and found the monooleic disaturated symetrical
volatiles onto a trap, and eluting trapped volatiles directly onto triglycerides most sensitive to the presence of nonchocolate
capillary GC-MS. Luminometry was applied by Tateo et al. fats. The presence of sesame oil was determined by Coors
(66G) to evaluate the storage stability and condition of citrus and Montag (14H) by HPLC analysis of sesamol, sesamolin,
oils. Evidence for protein-phenolic binding during coffee sesamin, and tocopherol with fluorometric detection. Farag
roasting was obtained by Trugo and Macrae (68G) by spec- et al. (20H) distinguished as low as 5% cow’s milk in buffalo
troscopic analysis and size distribution changes in extractables milk by regression analysis of GC fatty acid profiles of crys-
as measured by SEC-HPLC. tallized fats and mother liquor; myristic, palmitic, stearic, and
Chiang (15G) developed a sensitive HPLC procedure uti- oleic acid levels contributed to the equation. Gegiou and
lizing electrochemical detection to separate and quantitate Staphylakis (24H) determined the presence or absence of
capsaicins. Capsaicins can be detected in the presence of cocoa butter equivalents in chocolate from GC or GC/MS
piperine by changing the detector potential or by measuring analysis of triterpene alcohols. Gomez (26H) estimated
piperine at 340 nm UV. Piperine was measured by silica TLC mineral oil in vegetable oil at levels >0.1 % using TLC.
with UV densitometry by Jansz et al. (34G). Kawada et al. Mineral oil in vegetable oil has also been detected by Graciani
(36G) separated capsaicin from dihydrocapsaicin using re- et al. (28H) using HPLC separation of hydrocarbons isolated
verse-phase HPLC with electrochemical detection; the de- using silica gel chromatography. Gubman et al. (31H) assayed
tection limit was 12 pg. Krajewska and Powers (39G) bro- oil for the presence of phosphorus-containing compounds using
minated capsaicins with pyridinium bromide perbromide to a rapid and simultaneous reaction with molybdate and ex-
achieve reversed-phase HPLC separation of nordihydro- traction into chloroform followed by spectrophotometric
capsaicin from capsaicin. measurement. Theobromine in cocoa beans was quantitated
Kato and Sekikawa (35G) found direct injection of unstable by Hamann et al. (32H) using reverse-phase HPLC and
lactones derived from butter flavor onto GC-MS system phenacetin as internal standard. Hurst et al. (38H) measured
yielded best results. Moellering and Bergmeyer (49G) de- caffeine, theobromine, and theophylline in cocoa using mi-
veloped a specific enzymic method for detection of L-glucono-5 crobore reverse-phase HPLC and reported detection limits
lactone in tissue and foods. The lactone is converted to glu- ranging from 100 to 150 pg. Harland and Oberleas (33H)
conate-e-phosphate followed by oxidation with NADP+ and developed an analytical method to measure phytate in vege-
spectrophotometric measurement of the resulting NADPH. tables and grains which employs anion exchange chroma-
Lactones from ghee were isolated with 80-92% recovery by tography; the method was adopted as an official first action.
Wadhwa and Jain (72G) using a celite-digitonin-alumina Lemieux et al. (54H) compared three methods for phytate
chromatography column and acetonitrile elution. Collinge et measurement: ferric chloride precipitation, ion exchange
al. (16G) developed a reverse-phase HPLC method for the retention, and enzymatic hydrolysis. The ion exchange me-
analysis of glycyrrihizin and f?-glycyrrhetic acid in foods and thod was preferred for rapidity and low cost. Mazzola et al.
beverages. Abraham and Deman (1G) developed a rapid (61H) applied 31P FT NMR to the analysis of phytate in foods
infrared method for the determination of four isothiocyanates and found excellent agreement with published ion chroma-
in canola oil; results were confirmed by GC and GC-MS. An tographic methods. Il’ina et al. (40H) developed a technique
enzymic method for the determination of acetaldehyde using to measure carbon dioxide in noncarbonated beverages which
aldehyde dehydrogenase and spectrophotometric measure- utilizes both a gas electrode and a pH meter in tandem.
ment of NADH was conceived by Beutler (7G). Buetler (8G) Mustard seed coat presence was detected by Jamais et al.
worked out an enzymic method for ethanol determination by (41H) by colorimetric analysis of 4-hydroxyproline, after ox-
converting ethanol to acetaldehyde using alcohol de- idation to pyrrole by Chloramine T and reaction with Ehrlich’s
hydrogenase and NAD+ and then measuring acetaldehyde as reagent. Kikugawa et al. (48H) developed a chromogenic assay
described above. Cyclic sulfur compounds in shiitake for lipid hydroperoxides whereby reation of hydroperoxides
mushrooms, lenthionine, 1,2,4-trithiolane, and 1,2,4,5-te- with sesamol dimer in the presence of hemoglobin (metHb,
trathiane were determined using reverse-phase HPLC by Chen oxyHb, or carbonmonoxyHb) results in highly colored qui-
and Ho (14G). Dirks and Herrmann (19G) developed a re- nones or semiquinones; this technique was found to be more
verse-phase HPLC method for the analysis of phenolics, hy- sensitive than the peroxide value method. Krueger and
droxycinnamoylquinic acids, and 4-(/3-D-glucopyranosyl- Krueger (51H) assessed the purity of natural vanillin by ox-
oxy)benzoic acid in spices. Chlorogenic acid was the most idation with sodium chlorite to vanillic acid, decarboxylation
abundant and frequently encountered compound. Liquid by treatment with bromine, and analysis of released carbon
carbon dioxide extraction was utilized by Eberhardt and dioxide 13C/12C isotope ratio using MS. Lee et al. (53H)
Pfannhauser (22G) to isolate an elderberry monoterpene measured furfural and 5-hydroxyfurfural in citrus juice using
precursor, 3,7-diethyl-octa-l,5-dien-3,7-diol. Cinnamaldehyde reverse-phase HPLC and reported a 50 ppb lower detection
in cinnamon or cassia oil was determined by reaction with limit. Lever et al. (55H) realized 60 sample per hour turnover
p-hydroxybenzoic acid hydrazide and 5% sodium hydroxide for furfural analysis using a continuous flow system incor-
followed by spectrophotometric measurement at 352 nm by porating furfural conversion to hydrazone, dialysis, into
El-Obeid et al. (23G); benzaldehyde did not interfere. Volatile methanol, and fluorescence measurement of the zinc chealate.
aldehydes were converted to either thiazolidine or 2-acetyl- A reverse-phase HPLC method for furfurals in orange juice
thiazolidine via reaction with cysteamine at pH 6 or 8, re- was reported by Marcy et al. (60H) in which distillation of
spectively, by Hayashi et al. (28G) and then determined by the juice was employed as a cleanup step. Loeliger and Saucy
GC equipped with a thermionic N-P detector. Curcumin in (58H) applied chemilimunescence emmission to the deter-
spice mixtures was determined by Janssen and Gole (33G) mination of food autooxidation. By comparison with tradi-
by TLC resolution and conversion to rubrocurcumin with a tional peroxide value methods, the results from this technique
mixture of boric acid and oxalic acid followed by fluorometric were critically dependent upon the type of food being analyzed
scanning. Molnar-Perl et al. (50G) described an ion exchange and the presence of food additives (antioxidants). Muuse and
method for the rapid separation of Maillard reaction products, Van der Kamp (65 H) determined the presence of fractionated
deoxy fructosyllysines, and their pyridosine and furosine butterfat in natural butterfat by gravimetric analysis of sat-
hydrolysis products. Spiro and Price (64G) developed a urated triglycerides crystallized from hexane at 12.5 °C; this

230 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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technique was found superior to fatty acid analysis, analysis Morishita et al. (6422) isolated vegetable phenolics by prep-
of sn-2-fatty acids, and cholesterol analysis. Lipid identifi- arative reverse-phase HPLC and identified the components
cation was achieved by Nazer et al. (6722) by applying mul- using *H NMR and MS. Cheese quality has been assessed
tivariate statistical analysis to prolysis GC profiles. The from ultrasonic transmission and reflection measurements by
presence of spore-forming bacteria in canned, low-acid foods Northeved (6822). The extent of milk heat treatment was
(cream style corn and beef noodle soup) was confirmed by measured by Patel et al. (7322) using the methylene blue
Schafer et al. (7722) using GC analysis of butyric acid and reduction test for residual xanthine oxidase enzyme. Used
D-(-)-2,3-butanediol; this technique was adopted as official frying oil was detected from changes in the GC triglyceride
first action. As low as 4% palm stearin in palm oil was de- dimer peak shapes by Rogstad (7522). Takahashi et al. (7822)
tectec by Tan et al. (79H) by analysis of slip melting point, described an extraction, cleanup, and HPLC electrochemical
iodine value, and GC fatty acid and triglyceride composition. detection system for the carcinogenic compounds, amino-
Results were highly dependent upon the type of palm stearin imidazoquinoline and aminoimidazoquinoxal, in heated beef
fraction that was added. Tanner and Limacher (80H) mea- extract. Oyster freshness was determined by Tsunoda et al.
sured methanol, ethanol, and acetaldehyde in fruit juice by (8222) from residual succinate dehydrogenase activity as
addition of citric acid and direct injection into a GC containing measured by concurrent reduction of tetrazolium chloride to
a Carbopack B/6.6 CW column and FID detection. Wrigley red formazan.
et al. (84H) reviewed the utilization of electrophoretic patterns Patel (7222) reviewed competitive and noncompetitive im-
of grains and other foods to determine and characterize their munologic methods for determination of staphylococcal en-
genetic complement or similarity. A histochemical deter- terotoxins. Biru et al. (622) found ELISA simple and satis-
mination of glucosinolates in rapeseed oil or sectioned rape- factory for detection of staphylococcal enterotoxins in con-
seeds was developed by Yiu et al. (86H) where glucosinolates taminated foods. Notermans (6922) and Wieneke (8322)
are detected by reaction with 2V-2,6-trichloro-p-benzo- likewise described ELISA tests for staphylococcal enterotoxins
quinoneimine which produces a yellow color. to be accurate for assessment of foods associated with poi-
Quality of fruits and vegetables was determined by Akimoto sioning outbreaks. Hyde and Stahr (3922) correlated volatile
(222) using simultaneous quantitation of sugar and acid by trapping thermal desorption GC/MS results from mold-
13C NMR. Barroga et al. (422) measured polyphenbls in mung spoiled grains to the presence of mycotoxins. Carman et al.
beans using three methods (modified vanillin, Prussian blue, (922) developed a rapid GC-FID method to detect and
and protein precipitation) and found that results varied widely quantitate the plant toxin, myristicin, in fresh and frozen
according to mung bean treatment, rather than the method carrots.
used. Of the six rapid tests for frying oil quality, Croon et Casa et al. (2022) developed an immunoelectrophoresis
al. (15H) found the Foodoil Sensor and the Fritest tests most method to detect soluble horse muscle proteins in the presence
rapid and accurate. The country or origin of green coffee was of soluble bovine and procine muscle proteins. A test for
determined by Dyszel (19H) using pyrolysis MS. Com protein soluble pig muscle protein was also developed by Casa et al.
quality was related to the amide II to amide I infrared band (2 222). Garrone et al. (2322) detected hazelnut presence in
intensity ratios by Fedenko (2222). Geilinger et al. (2522) found chocolate by thin-layer electrophoresis of ammonium sulfate
that changes in chocolate because of processing variations protein extracts on cellulose acetate. From proline, aspartic
could be attributed to changes in the integrity of cocoa starch. acid, and alanine levels, Greenberg and Dower (2922) were able
Hill and Gasson (35H) developed a SDS-polyacrylamine gel to detect added whey protein in nonfat dry milk powder,
profiling technique to determine the occurrence of casein regardless of heating. An ELISA test to detect soya protein
hydrolysis by S. lactis proteases in milk. A headspace GC in meat was developed by Griffiths et al. (3022). Janssen et
method for determination of ethanol in the aqueous phase of al. (4222) detected soya proteins in heated meat by applying
canned salmon was deeloped by Hollingworth et al. (3622) and a selective immunoperoxidase system to blots derived from
adopted as official first action. Ohhashi (7022) published a SDS polyacrylamide gels; antigenicity was decreased in the
freshness index equation for fish based on levels of ATP, ADP, presence of liver at 25 °C. Jones and Patterson (4322) de-
AMP, IMP, inosine, and hypoxanthine. By profiling various veloped an indirect ELISA test for unheated meat species
protein extracts from different species of wheats, Huebner identification that uses crude antisera rather than highly
and Bietz (3722) were able to demonstrate relationships be- purified antibodies. Krause and Belitz (5022) differentiated
tween species and profiles that are useful for quality control milk from different species by isoelectric focusing of their
and breeding selection. Kamarei and Karel (4422) measured Y-2-caseins. Muscle protein presence in mixtures with soya
the extent of autooxidation in freeze-dried meats by protein and rind was achieved by Lindberg et al. (5722) hy
fluorescence; excitation and emission wavelength maxima were reverse-phase HPLC profiling of dansylated amino acids and
different in extracts containing oxidized components. Nakhost application of multivariate statistical analysis. As low as 0.5%
and Karel (6622) developed a myoglobin-based indicator soya protein was detected in meat, cooked or uncooked, by
system for freeze-dried meat autooxidation in which myoglobin Ravestein (7422) using an ELISA test; lower detection limits
insolubility, metmyoglobin concentration, and dimer to mo- could be achieved by using immunoblotting techniques.
nomer ratio are assessed. Grain spoilage by microorganisms Yehchen and Hsu (8522) detected protein hydrolysate in soy
was assessed by Kaminski et al. (4522) by colorimetric mea- sauce by measuring the levulinic acid content by HPLC with
surement of the volatile carbonyl acetone equivalents as ox- bromocreosol purple postcolumn detection.
imes. The thiobarbituric acid method for rancidity in fish Demmer and Werkmeister (2 722) found that extracts from
was improved by Ke et al. (46H) by incorporating acid hy- frozen pork and not fresh pork contained the mitochondrial
drolysis and distillation steps prior to reaction with 2-TBA enzyme, /3-hydroxyacyl-Coenzyme A dehydrogenase. Fresh
and spectrophotometry at 538 nm. Kneifel and Ulberth (49H) liver was discriminated from frozen liver by Gottesmann and
evaluated the extent of milk heat treatment by various casein Hamm (2722) using the marker enzyme, /3-hydroxyacyl-Co-
precipitation procedures and SEC-HPLC profiling of milk enzyme A dehydrogenase; results were not definitive for chilled
proteins. Ammonia, a quality indicator substance for squid, liver samples. Salfi et al. (7622) differentiated between fresh
was measured by LeBlanc and Gill (5222) by amination of and frozen fish by separating aspartate aminotransferase
a-ketoglutaric acid in the presence of glutamate dehydrogenase isozymes and staining for enzyme activity.
and NADH, and concurrent oxidation of tetrazole to colored Chen and Lin (2322) measured the 14C content of plants by
formazan. Lin and Cousin (5622) measured the OPA- using Aquasol-2 as liquid scintillant and a low background
glucosamine derivative by reverse-phase HPLC with scintillation counter. Derbesy (2822) reviewed the practical
fluorescence detection to estimate the extent of mold con- uses of stable isotope analysis for quality assessment of fla-
tamination of processed foods. Manz (5922) determined the voring materials; 14C,13C/12C, 2H/1H, and 180/160 ratios were
presence of heat-denatured bovine and porcine proteins using taken on vanillin and anethole. Palagyi (7222) measured 131I
an ELISA test developed for the a-2-globulin fraction. Ra- in drinking water by entrapment on a polyurethane column
peseed maturity (quality) was assessed by Minkowski and impregnated with a terf-alkylamine-I complex and taking
Schubert (6222) from the optical density of chlorophyll at 669 counts on the affinity column directly. To and Rack (8222)
nm in a petroleum ether-ethyl ether extract. Using o-solanine measured 131I in formaldehyde-preserved milk by first sepa-
as an antigenic hapten, Morgan et al. (6322) developed an rating protein and liquid phases and then oxidizing I' to iodine,
ELISA test for potato glycoalkaloids which gave good cross extracting with carbon tetrachloride, reducing iodine to 1“ with
reactivity with a-solanine, a-chaconine, and demissine. HS03“, and precipitating Pdl for both phases.
ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 »231R
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Boegl and Heide (7H, 8H) determined foods that had been Sample preparation and mineralization techniques for el-
treated with y irradiation or ultraviolet irradiation using emental analysis have been emphasized in some publications.
chemiluminescence, y irradiation of spices was measured by Microwave oven digestion techniques were reported by Ishii
Heide and Boegl (34H) by measuring the chemiluminescence (43J) and by White and Douthit (105J) who tested the
of a mixture of the spice with luminol solution. Light yield technique with NBS reference materials and found good
and the time-course of emission were different for each spice, agreement for the elements Ba, Ca, Mg, Mn, P, K, Na, S, and
and oxygen concentration, pH, and luminol freshness affected Zn using ICP. DeMura et al. (20J) report a microwave oven
the results. Kiel (47H) developed a microchemiluminescent digestion method for the determination of Zn, Cu, Mn, Pb,
probe material to assess peroxidative, heat, and ionizing ra- and Cd in food and reported recoveries of 84-113%. Gillain
diation damage to cells by making a glutaraldehyde cross- and Rutagengwa (29J) compared two ashing methods for the
linked complex from serum albumin, a heme protein, and a determination of Zn, Cd, Pb, Cu, Sb, and Bi in milk by dif-
luminescer. ferential pulsed anodic stripping polarography and found the
Acton et al. (1H) determined the structure of protein gels Teflon bomb ashing method gave satisfactory results for the
using ultrasound imaging. A screening technique for lectins more volatile trace elements. A preparative digestion pro-
in vegetables and spices was developed by Andersen and cedure for biological and diet samples prior to the analysis
Ebbesen (3H) which utilizes line-drive immunoelectrophoresis of essential minerals by AAS was described by Hill et al. (38J).
and hemadsorption. Bianchini et al. (5H) determined the Murphy et al. (66J) compared wet ashing and flame/furnace
sterol and triterpene composition of sunflower oil by HPLC aerosol sampling techniques for the determination of Cd, Cu,
analysis of their acetates. Free formaldehyde was determined Mn, Ni, and Cr in food by AAS. An automated wet digestion
in meat by DeFreitas et al. (16H) by extraction, distillation procedure for the determination of As, Cd, Cu, Hg, Pb, Se,
in the presence of sodium sulfate and phosphoric acid, and and Zn in animal tissue was described by Salisbury and Chan
photometry. Lysinoalanine was detected and quantitated in (81 J). A procedure including sampling, transport, drying,
milk proteins by Fritsch (22H) by ion exchange chromatog- homogenization, digestion, and voltimetric measurement for
raphy and postcolumn reaction with OPA and fluorescence the determination of Zn, Cd, P, Cu, Ni, and Co in meat of
detection. slaughtered cattle was described by Narres et al. (70J). A
review of the certification of reference materials by the Na-
MINERALS tional Bureau of Standards was published by Alvarez (3J) and
Trends in methodology continue to favor multielement Miller-Ihli and Wolf (62J) described the characterization of
techniques. The application of inductively coupled plasma NBS SRM 8431 for 17 elements.
atomic emission spectrometry (ICP-AES) techniques also Aluminum and iron in food samples and water were de-
continue to increase. Review articles pertaining to mul- termined by Yan and Schwedt (109J) using ion chromatog-
tielement analysis consist of the following: Mannino (58J) raphy and postcolumn derivatization. McCabe and Ottaway
who reviewed electrochemical techniques for the determina- (60J) described a novel method for the determination of As,
tion Pb, Cu, Zn, Cd, As, and Hg in foods; Herrador et al. (36J) Sb, and Se in animal food protein as their hydrides using AAS.
who compiled spectrophotometric methods for Pb, Hg, Cd, Oxygen bomb combustion and automated AAS with hydride
Se, and As in milk; and Boyer et al. (15J) who examined the generation was used by Narasaki (68J) for the determination
interlaboratory variability in trace element analysis of col- of As and Se in butter and polythenes. Muenz and Lorenzen
laborative studies conducted by the AOAC. (65J) described a procedure for the selective determination
A method for the determination of Mn, Fe, Cu, and Zn in of inorganic and organic arsenic in foods by AAS using the
tropical foods was reported by Benzo et al. (13J) who digested hydride generation technique. Methods for the determination
the samples with HNO3/H2SO4/H2O2 followed by AAS. of As in sugar, beet pulp, and molasses by AAS after hydride
Puchyr and Shapiro (78J) described a procedure for extraction formation were studied collaboratively by Huijbregts et al.
and determination of Al, Fe, Sn, Zn, Ca, Mg, Ni, Cu, Cr, Ca, (40J) and compared to neutron activation and absorptiometric
and K in foods by AAS. Trace elements were determined in methods. Arsenic in food samples by AAS with graphite
milk by AAS, neutron activation analysis, and photon-induced furnace was reported by Dabeka and Lacroix (19J) who de-
X-ray emission by Gharib et al. (28J). Kluessendorf et al. scribed a routine method involving dry ashing, by Okubo et
(47J) described a procedure for Pb, Cd, and Zn in livers by al. (73J) who used nickel ion with wet digestion, and by
Zeeman AAS of solid samples. Knezevic and Kurfuerst (48J) Tsukada et al. (96J) who reported a rapid microwave oven
compared solid sampling and digestion techniques by gra- digestion method. A spectrophotometric method for the
phite-tube AAS for the determination of Cd, Pb, Hg, Cu, and determination of boron in natural waters and vegetable matter
Cr in food packaging materials and found the values obtained was described by Aznarez and Mir (7J). Ochiai et al. (72J)
comparable. Yan et al. (108J) found comparable results by determined boron in foods by dry ashing, dissolving the ash
ion chromatography (IC) and AAS for the determination of in dilute HC1, and chromatography on a cation-exchange resin
Cu, Zn, Fe, Mn, Ca, and Mo in beverages; the IC method column and fluorescence detection.
shows no matrix effect and unlike AAS is suitable for si- Analytical methods for the determination of Ca and Mg in
multaneous elemental analysis. A method for the determi- wine were reviewed by Baluja-Santos et al. (9J). Kindstedt
nation of Fe, Cu, Mn, Co, Cr, Mo, and Ni in milk by elec- et al. (46J) described an improved complexometric method
trothermal AAS was described by Mingorance and Lachica for the determination of calcium in cheese using hydroxy-
(63J). Simultaneous determinations of nine elements in a naphthol blue as the indicator with EDTA titration. Calcium
reference standard and fruit juice were reported by Harnly and potassium were determined in skim milk powder by Al
(35 J) who used a commercially available aerosol deposition- Hitti et al. (1J) who used a dry ashing procedure with ion-
carbon furnace atomizer with a multielement atomic ab- selective electrodes. Alvarez de Eulate, et al. (2J) determined
sorption continuum source spectrophotometer. Evans and Cd, Cu, and Pb in food grade salt by extraction of the am-
Read (22J) determined Li and Rb in foodstuffs using flame monium pyrrolidinedithiocarbamate complex with 4-
atomic emission and absorption spectrometry. ICP methods methyl-2-pentanone followed by AAS. Direct solid sampling
are being used more commonly and the following have been analysis of Cd, Pb, and Hg in fresh seafood by Zeeman AAS
reported for food analysis: Lyons et al. (56J) described a with graphite furnace was described by Grobecker et al. (31 J).
procedure for the determination of Ca, Zn, Mn, Fe, Mg, and Narres et al. (69J) determined Cd and Pb in milk directly with
P in feed and plant material; Shiraishi et al. (86J) applied the platform furnace Zeeman AAS. Olayinka et al. (74J) devel-
technique to the direct multielement analysis of total diet oped a slurry technique for the determination of Cd in dried
samples for major, minor, and trace elements; and Takeo (95J) foods by electrochemical atomization AAS. Cobalt was de-
described instrumentation and methodology to determine termined in foods by Barbera et al. (11J) after wet digestion,
minerals and trace elements in tea. A collaborative study for liquid extraction, and AAS. Copper was determined in milk
the determination of Ca, Cu, Fe, Mg, Mn, P, K, Na, and Zn powder by Khammas et al. (45J) by electrothermal atomic
in infant formulas was reported by Suddendorf and Cook (94J) absorption and atomic emission spectrometry. Chromium was
and the ICP method was adopted official first action by the determined in meat samples by Schindler (83J) who digested
AOAC. Nickdel and Barros (71 J) described a procedure for the sample with HN03 followed by graphite tube AAS with
major and trace elements in citrus juices using an automated Mg(N03)2 added as a modifier. Farre et al. (23J) reported
fast sequential multielement spectrometer combined with an AAS method for the determination of Cr in foodstuffs in
ICAP-AES. which the organic matter is digested with HN03, followed by

232 R« ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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oxidation to Cr(Vl) and extraction with methyl isobutyl ke- used Hyamine-2389 and appropriate electrolytes to separate
tone. Sn and Pb peak potentials to allow the simultaneous deter-
Iron in wines, foods, and minerals was determined by salinas mination of these elements. Titanium in milk powders was
et al. (80J) using a new reagent-5,5-dimethyl-l,2,3-cyclo- determined by ICP-AES by Van Betteray-Kortekaas et al.
hexanetrione 1,2-dioxime 3-thiosemicarbazone and absorbance (100J) after heating at 450°, fusing with K2S207, and dissolving
measured at 550 nm. Singh et al. (89J) proposed l-(2- in a hot H2S04 solution. Zinc was determined in milk by Singh
quinolylazo)-2,4,5-trihydroxybenzene as a sensitive chromo- et al. (88J) using l-(2-quinolylazo)-2,4,5-trihydroxybenzene
genic reagent for the simultaneous determination of Fe and as a spectrophotometric reagent. A selective spectrofluoro-
Cu in foodstuffs, body tissue, and milk samples. Viladrich metric determination of zinc with 5,7-dibromo-8-quinolinol
Gonzalbez et al. (102J) compared AAS and ICP for the de- and its application to food samples was described by Fer-
termination of Fe and Cu in fats. A method employing isotope nandez et al. (24J). An AAS method for the determination
dilution fast atom bombardment mass spectrometry was de- of trace amounts of zinc in canned juices after ion exchange
veloped by Gharaibeh et al. (27J) for the quantitative de- separation was reported by Aziz-Alrahman (6J).
termination of Fe in foods. A rapid method for the deter- Multiple simultaneous anion analysis was reported by
mination of Pb in milk has been proposed by Mannino and Schmuckler et al. (84J) who separated H2P04~ CT, Br', N03',
Bianco (57J) who used potentiometric stripping analyses I', and S042' in fruit juices on a reversed-phase C18 chroma-
without sample pretreatment. A method for the determination tographic column. Total bromine in food was determined by
of Pb in spinach involving minimum sample preparation was Oyamada et al. (75J) who ashed the sample, reacted the lib-
described by Stephen et al. (92J), who used electrothermal erated bromine with styrene monomer, and determined the
AAS. Andersen (4J) described a screening method for the resulting bromostyrene by gas chromatography with electron
determination of Pb in infant formulas and powdered milk capture detector (GC-ECD). Stijve (93J) also used GC-ECD
using Zeeman corrected graphite furnace AAS. A graphite to determine inorganic bromide in foods by reaction with
furnace AAS method for the determination of Pb and Cd in propylene oxide and found the results to be comparable to
foods after nitric-perchloric acid digestion and coprecipitation the ethylene oxide method. Nangniot et al. (67J) compared
with APDC was reported by Dabeka and McKenzie (18J). bromide residues in vegetables by gas chromatography and
Cold-vapor AAS was used by Koops et al. (50J) for ultratrace by ion selective electrode after separation of other halides by
amounts of Hg in milk products and by Coles et al. (16J) who ion-exchange chromatography and preferred the latter method
described a reference method in detail for the determination because of accuracy, rapidity, and simplicity. Mori et al. (64J)
of Hg in foods. A method of digestion by using a mixture of used a postcolumn derivatization procedure to determine
HC1, HN03, and H2S04 was developed by Louie et al. (55J) potassium bromate in food by HPLC. Residual chlorine in
for the determination of Hg in a wide range of food products foods due to NaOCl or bleaching powder was determined by
with a detection limit of 0.01 mg/kg using cold-vapor AAS. Hidaka et al. (37J) who used headspace gas chromatography
Mercury loss during pretreatment and digestion of plant after conversion of the chlorine to cyanogen chloride. The
materials was investigated by Semu et al. (85J) who found Volhard and potentiometric methods for the determination
that digestion with HN03/KBr03 resulted in a recovery of of chloride in meat products were compared in a collaborative
90-100% of the added Hg. Grobenski et al. (32J) developed study reported by Beljaars and Horwitz (12J). Balulescu (10J)
a method for the determination of Hg by Zeeman graphite determined salt in foods using an automated ion-selective
furnace AAS and successfully applied it to several environ- electrode method at a rate of 40 samples/hour and measuring
mental samples. Kruse (52J) reviewed mineralization methods Cl, Na, and K. Johnson and Olson (44J) made a comparison
for the determination of Hg, Cd, and As in fish products by of available methods for the determination of salt in cheese
AAS. Methods for the determination of total Mo and V in including the Mohr, Volhard, ion selective electrode, and
foods were described by Evans and Caughlin (21 J) who chloride analyzer procedures. Matsumoto et al. (59J) de-
measured Mo at 680 nm as the toluene-3,4-dithiolate complex scribed a conductometric flow injection analysis method for
and V at 415 nm by its catalytic effect on the oxidation of gallic the determination of salt in food products at a rate of 70
acid with peroxymonosulfuric acid. Kohiyama et al. (49J) samples/hour. Singer and Ophaug (87J) described methods
found that extraction with HC1 is suitable for the routine for acid diffusable and total FI content in food.
analysis of nickel in hydrogenated oils using AAS. Vaeth and Total iodine in foods was determined by an automated
Holz (99<J) determined trace amounts of Ni and Fe in fats and colorimetric method based on iodide-catalyzed reduction of
oils by AAS with a reduced sample treatment procedure. cerium by Fischer et al. (25J). Wiechen and Kock (107J)
A validation study was reported for a fluorometric method developed a routine method for the determination of low
for the determination of Se in food by Vaessen et al. (98J) iodine concentration in milk based on the Sandell-Kolthoff
who decomposed the samples in a closed system and com- reaction. A new method for the determination of I in food
plexed the Se(IV) with 2,3-diaminonaphthalene. A wet de- samples by isotope dilution mass spectrometry was described
composition procedure with a mixture of HN03, HC103, and by Schindlmeier and Heumann (82J) who decomposed the
HC104 was described by Irsch et al. (42J) for the determi- organic matrix with a mixture of acids and isolated the I by
nation of Se in samples of plant and animal origin using the extraction with CC14. Lawrence et al. (53J) determined iodide
hydride-AAS method. A sodium ion selective electrode me- in table salt by X-ray fluorescence after absorbing the HI
thod has been reported by Fulton et al. (26J) for the deter- generated from the sample onto an ion-exchange resin loaded
mination of sodium directly in processed meat products. A paper disk. The determination of I radioisotopes in milk has
slurry method for direct flame emission determination of Na been improved to increase accuracy and sensitivity and to
and K in processed meat and pizza products was reported by shorten analysis time by Bettoli et al. (14J) who concentrated
Wichman et al. (106J) who used a sonic/cavitational homo- the I isotopes by anion-exchange chromatography on AgCl-
genizer for sample preparation. Tin was determined in canned Si02 and then determined them by iodide-specific membrane
foods by Aznarez et al. (8J) using AAS and hydride generation electrode and y and counting. Iodine radionuclides in milk,
in a nonaqueous medium. Hocquellet (39J) described the vegetables, and fish were also determined by Meloni et al.
direct determination of tin at ultratrace levels in fats and oils (61J) who used 7-ray spectrometry for 126I and 131I and liquid
by AAS with electrothermal atomization. Total tin deter- scintillation counting for 129I. The well-known Kroller method
minations were reported by Krull and Panaro (51J) using for the determination of cyanide was modified by Van Eeden
continuous on-line hydride generation followed by dc plasma et al. (101J) who replaced the harmful chemicals benzidine
emission spectroscopy (HY-DCP); organotin analysis and and Br with barbituric acid and N-chlorosuccinimide and
speciation can be accomplished by interfacing this HY-DCP applied the method to human foods and animal feeds.
setup with HPLC. Gutierrez et al. (34J) described a rapid Nitrate-N in vegetables was determined by Hunt and
extractive spectrophotometric method for the determination Seymour (41 J) using an automated anion-exchange HPLC
of Sn in canned foods with 5,7-dichloro-8-quinolinol. A technique. Vlacil and Vins (103J) determined N03' in milk
fluorometric method for the determination of Sn at the na- products and milk-based infant formula using liquid chro-
nograms per milliliter level in canned beverages was reported matography on Spheron and direct photometric detection at
by Rubio et al. (79J) who reacted Sn with diacetylmonoxine 205 nm. Nitrate and nitrite in food products were determined
nicotinylhydrazone to form a fluorescent complex. The de- by Unger and Heumann (97J) using isotope dilution mass
terminations of Sn and Pb in canned fruit juice by differential spectrometry; 15 N03' and 15 N02' spikes were used for the
pulsed polaragraphy were reported by Guinon et al. (33J) who dilution technique and negative thermal ions measured in the

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 233 R
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mass spectrometer. Nitrate and chloride ions in food were meat and nonmeat products was developed by Ali (1L).
determined by Penchuck et al. (77J) using a single-column Ashoor et al. determined acetic acid in vinegar and other foods
ion chromatography technique. A procedure to measure P by HPLC on an Aminex HPX-87H ion exchange column (2L)
content of soybean and com oil has been developed by Sinram and citric acid in a variety of foods using the same column
(90J) using the relationship between P level due to phos- with a micro-guard ion-exclusion cartridge and UV detection
phatides in vegetable oil and turbidity formed in phosphatide at 210 nm (3L). Badoud et al. (4L) reported an HPLC method
mixtures; P vs. turbidity data formed nearly linear relations for the analysis of the main carboxylic acids in coffee, wine,
for crude, degummed, once-refined, bleached, and deordorized and fruit juices using conventional and small-bore columns.
soybean and corn oil process samples. List et al. (54J) de- Boehme et al. (8L) analyzed bread organic acids by HPLC.
termined phosphate and S02 in fruit juices and wine by flow Chikamoto et al. (10L) simultaneously determined ethanol
injection analysis and photometry; a gas diffusion cell was used and propionic acid in bread and cake mixes by GC analysis
to eliminate interference for the determination of S02. Warner of the steam distillates of samples. Rabe (49L) determined
et al. (104J) reevaluated the Monier-Williams method for sorbic acid in bread and bakery products by colorimetric, TLC,
determining sulfite in food and reported recovery values for GC-FID, and HPLC on RP-8 or RP-18 columns. The merits
a variety of commercial food products. Results of a 10-min of the different methods were discussed.
flash distillation and 10-min ion chromatography determi- Chonan (11L) determined steam distillable propionic, bu-
nation compared favorably with the results from the con- tyric, caproic, and caprylic acids in milk by reversed-phase
ventional Monier-Williams method for total sulfite in foods HPLC of their p-bromophenacyl esters with UV monitoring
according to Anderson et al. (5J). Sulfur on grapes and wheat at 254 nm.
was determined by Goewie et al. (30J) using a liquid chro- Coppola et al. (12L) reported the results of a collaborative
matograph with UV and electrochemical detection. study of a reversed-phase HPLC method for the determination
A fluorometric reaction rate method for determining hy- of quinic, malic, and citric acids in apple juice and cranberry
drogen peroxide at the nanomolar level was proposed by juice cocktail. Analyses were performed using two reversed-
Peinado et al. (76J) and applied to the analysis of coffee, tea, phase columns, phosphate buffer (pH 2.4) mobile phase, and
and milk samples; the method is based on the Mn(II)-cata- UV detection at 214 nm. The method has been adopted
lyzed oxidation of 2-hydroxynaphthadehyde thiosemi- official first action by AOAC. Gancedo et al. (15L) analyzed
carbazone. Srinivas et al. (91J) described a method for the oxalic, citric, galacturonic, malic, and pyrroleindone carboxylic
continuous determination of H202 in food based on the cat- acids in tomato juice by HPLC. Sorbic acid in jam and
alytic decomposition of H202 by LaCo03; the 0 liberated is marmalade was determined by reversed-phase HPLC and UV
measured by a gas measuring buret. A method has been detection as reported by Goto et al. (17L). Hayakawa et al.
developed to determine headspace 02, N2, and C02 in bev- (18L) determined the oxalic acid content of spinach by the
erages and packages by Cook et al. (17J) using headspace gas HPLC of defatted aqueous extracts using a strong anion-ex-
chromatography. change column with phosphate buffer (pH 4.1) as mobile phase
and UV detection at 202 nm. Oxalic acid was also determined
MOISTURE in spinach by Kok et al. (26L) using ion-interaction re-
Books that are of interest to analysts involved with moisture versed-phase HPLC and amperometric detection and by
measurements are Aquametry. Part 2, Electrical and Elec- Ohkawa et al. (42L) in spinach and beverages by GC analysis
tronic Methods: A Treatise on Methods for the Determi- of the dimethyloxalate ester.
nation of Water" (12K) and Moisture Sorption: Practical Lin et al. (30b) analyzed wine and juice carboxylic acid by
Aspects of Isotherm Measurements and Use (8K). HPTLC on cellulose; developed chromatograms were stained
Bussiere et al. (IK) determined the water activity (aw) of with xylose-aniline reagent and densitometrically scanned at
confectionery products using various calculation methods. 546 nm.
Unsaturated solutions of sodium chloride were proposed by Mattiuz et al. (35L) determined short chain (C!-C4) organic
Chirife et al. (3K) for calibrating hygrometers for the deter- acids in nutritive sweeteners as table sugar and high-fructose
mination of aw. Gerschenson et al. (5K) discussed the in- corn syrup using ion-exclusion HPLC coupled with suppressed
fluence of organic volatiles such as 1,2-propylene glycol on aw conductivity detection. Mazzola et al. (36b) analyzed phytate
measurements with fiber-dimensional hygrometers. Theo- in foods using an ion chromatographic method and a 31P
retical prediction of aw of several standard saturated salt Fourier transform NMR spectrometric method. Mentasti et
solutions agreed well with experimental measurements for al. (37b) reported a method on derivatization, identification,
most salts studied at 15° and 35° degree according to Kitic and separation of carboxylic acids in wines and beverages by
et al. (6K). Measurements of aw of salt solutions and foods HPLC.
by several electronic methods were compared by Stamp et al. Matsuo et al. (34b) analyzed soy sauce for benzoic acid and
(13K) to direct vapor pressure measurements; measurement levulic acid by TLC on kiesel gel 60 F-254 plates and by gas
of aw of foods gave values differing by an average of 0.051 aw chromatography on a 10% DEGS + 1% H3P04 on Chromo-
units compared with VPM readings. A simple dew-point sorb W (AW DMCS) 60-80 column. Benzoic acid and p-
hygrometer which injects water into a sample gas and its hydroxybenzoate esters were simultaneously determined in
application for determining moisture in foods by measuring soy sauce using reversed-phase HPLC and UV detection at
the relative humidity differences were reported by Ueda et 256 nm, as reported by Otsuka et al. (43b). Panari (44b)
al. (14K). simultaneously determined pyruvic, succinic, lactic, formic,
Sanna (11K) reported the results of a collaborative study acetic, propionic, and pyroglutamic acids in cheese by HPLC
to revalidate the reliability of AO AC method 30.005 for the on an Aminex-HPX 87H column at 42° with 0.013 N sulfuric
determination of moisture in spices by distillation; for most acid as mobile phase and UV detection at 220 nm. Picha (45b)
spices, the method presented in this study was acceptable, developed an HPLC technique for the analysis of malic, citric,
whereas red pepper produced less satisfactory results. Rueegg succinic, and trace amounts of oxalic and oxalacetic acids in
et al. (10K, 9K) made a comparison of four commercially raw and baked sweet potato. Chromatography was carried
available Karl Fischer (KF) reagents and compared the water out using an Aminex HPX-87H cation-exchange column at
content in milk and milk products as determined by KF 75° with 0.0008 N H2S04 as mobile phase and detection at
titration and reference oven drying method. A Karl Fischer 214 nm. Malmberg et al. (32b) determined chlorogenic acid
method using a high potency KF reagent was applied by in potato tubers by reversed-phase HPLC with UV detection
Koizumi et al. (7K) to the determination of moisture in at 313 nm and by GLC of silyl derivatives of chlorogenic acid
high-moisture foods. Pulsed NMR was used by Gambhir et or of quinic acid. Results of chromatographic methods (HPLC
al. (4K) for the simultaneous determination of moisture and + GLC) were compared with the results of a spectrophoto-
oil in oilseeds. Chin et al. (2K) reported the results of a metric method and an enzymic hydrolysis method.
collaborative study using microwave oven drying for the de- Puttemans et al. extracted organic acids from food matrices
termination of total solids in processed tomato products and by ion-pair formation with tri-n-octylamine and after back
the results agreed well with the vacuum oven drying method. extraction to an aqueous phase, simultaneously determined
synthetic dyes, benzoic acid, sorbic acid, and saccharin in soft
ORGANIC ACIDS drinks and lemonade syrups (47b) and sorbic acid, benzoic
A reversed-phase HPLC method for simultaneous deter- acid, and saccharin in yogurt (48b). Rocklin et al. (50b)
mination of benzoic acid, sorbic acid, and four parabens in separated and detected organic acids in wine and milk by ion

234 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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chromatography with chemical suppression and conductivity NITROGEN

detection. Tsuji et al. (54L) separated quinic, malic, and Osborne (78M) wrote a comprehensive review comparing
shikimic acids in plum fruit using cation-exchange HPLC and and contrasting all chemical and physical methods for
UV detection at 210 nm. measuring the protein content of cereals. Aiyar et al- (2M)
An empiric formula for determination of lactic acid in optimized the Kjeldahl procedure for commercial lactalbumin
starter cultures in reconstituted skimmed milk on the basis by incorporating a 24-h preincubation with sulfuric acid and
of its pH was reported by Joex et al. (21L). Kogure et al. (25L) using mercuric oxide rather than copper sulfate catalyst. Use
determined the acid content of fruit juices by treating the juice of the oxidant peroxymonosulfuric acid was claimed by Hach
with NaHC03 or its analogues and measuring the C02 pro- et al. (43M) to preclude the use of metal catalysts and salts
duced by the reaction. Cereda et al. (9L) determined formic, in the Kjeldahl procedure; colorimetric detection was achieved
acetic, propionic, butyric, lactic, and succinic acids in fer- by an improved Nesslerization reaction. Horvath et al. (47M)
mented cassava starch by silicic acid chromatography. Fujita compared Kjeldahl and near-IR techniques for protein mea-
et al. (14L) used DEAE-cellulose column chromatography and surement in the presence of nonprotein nitrogen (phenyl-
UV absorbance at 325 nm for the estimation of chlorogenic alanine) and cellulose; near-IR yielded superior results for
acid in pear and apple fruits. Matsumoto et al. (33L) in- protein in the presence of phenylalanine. Kjeldahl analysis
vestigated conductometric flow injection analysis for meas- digestion step for animal feed was reduced by Kane (55M)
uring the organic acid content of fruits and Murakami et al. using a catalyst composed of copper sulfate and titanium
(40L) colorimetrically determined sorbic acid in a wide variety dioxide. Venter et al. (103M) optimized the semimicro
of foods in the presence of ethanol. Kjeldahl protein procedure for powdered dairy products by
Coffee nonvolatile acids were determined by Engelhardt scrutinizing catalysts, sample size, digestion procedure, and
et al. (13L) using capillary isotachophoresis and capillary gas distillation procedure.
chromatography. The GC method was the most sensitive and Bender (9M) reviewed 50 methods for protein quality de-
is the method of choice for exact work. Capillary isotacho- termination. Abd El-Salam et al. (1M) demonstrated quan-
phoresis was more rapid and reproducible and may be used titative differences between species using,a single calibration
in screening studies. Kikunaga et al. (24L) determined free on the Milkoscan 104 A/B infrared milk analyzer. Sjaunja
and total oxalic acid, oxalacetic acid, a-ketoglutaric acid, citric and Andersson (93M) evaluated a new infrared milk analyzer,
acid, and succinic acid in spinach using capillary isotacho- the Milko-Scan 605. Ameth (4M) described a continuous flow
phoresis. Prochazka et al. (46L) determined formic, acetic, apparatus that determines protein by reaction of trinitro-
propionic, butyric, valeric, and caproic acids in molasses using benzenesulfonic acid with an acid hydrolyzate and measure-
an automatic isotachophoresis analyzer. ment at 420 nm. Maximum protein solubilization from peas
Several enzymic-spectrophotametric methods for deter- was found by Banu et al. (6M) to be dependent upon particle
mining food organic acids were published. Bergmeyer et al. size, solvent/flour ratio, ionic strength, and pH. Findlay et
(5L) discussed a method for determination of acetate in foods al. (31M) characterized meat tenderness by analysis of dif-
like wine and fruit juices by using acetate kinase, phospho- ferential scanning calorimetry (DSC) heating thermograms
transacetylase, citrate synthase, and malate dehydrogenase with respect to endothermic reaction order and number of
following sample pretreatment for removal of 2-oxoglutarate, cooperative units participating in the transition. Ihekoronye
pyruvate, and lactate. Beutler (7L) determined acetate in (49M) employed trinitrobenzenesulfonic acid to monitor the
beverages using acetyl-CoA synthase, citrate synthetase, and reaction rate of enzymatic hydrolysis of proteins using various
malate dehydrogenase and sorbic acid (6L) in foods and fruit protease mixtures and protein substrates. Keller and Neville
preserves. A procedure for determining L-(+)-tartrate was (56M) compared four colorimetric procedures, biuret, Low-
described by Giffhorn et al. (16L). Oxalate was determined ry-Peterson, Bio-Rad Coomassie Blue, and Pierce BCA, for
in beer and fruit juices by oxalate oxidase, catalase, and al- determining the protein content of human milk; correlations
dehyde dehydrogenase as reported by Heinz et al. (20L). with micro-Kjeldahl were 0.96,0.97,0.89 and 0.99, respectively.
Kayahara et al. (23L) determined acetic, succinic, and citric Amide nitrogen in sodium carbonate extracts of food products
acids in soybean pastes. Lagemann et al. (27L) analyzed cocoa was determined by Paredez-Lopez and Guevara (80M) using
and cocoa products for oxalic acid. Moellering (38L) discussed an ammonia-specific electrode. The presence of soy protein
citrate analysis in liquid and solid foods. Schaller et al. (51L) in meat was determined by Raghavan et al. (86M) by pyro-
oxidized formic acid to C02 by NAD in the presence of formate lysis/GC and quantitation of o-methyoxyphenol, dimeth-
dehydrogenase and measured the formad NADH spectro- oxyphenol, and 2,3-dithiabutane. In a comparison of Kjeldahl,
photometrically. biuret, and near-IR methods for rapeseed protein content,
Healey et al. (19L) determined formic, acetic, propionic, Bengtsson (10M) found that near-IR was more convenient to
and butyric acids in cane molasses by GC analysis of their use routinely and yielded comparable data. Honigs et al.
benzyl esters on a column of 20% DEGS on Chromosorb W. (46M) developed a mathematical algorithm for the discri-
Katagiri et al. (22L) GC determined lactic, citric, succinic, and minant selection of unique sample spectra which improved
pyroglutamic acids in miso. Lamkin et al. determined pro- calibrations based on selected training sets. Near-IR wave-
pionic acid in corn (28L) and in grain sorghum (29L) by GC. length calibrations were developed by Kaffka and Martin
Littmann (31L) determined nonvolatile acids in shell eggs and (54M) for protein determinations in “animal protein meal”.
sugar-treated liquid eggs by GC of methyl esters on packed Valdez and Summers (102M) found protein determinations
and capillary columns and found that microbial spoilage of of poultry muscle tissue to be more precise with near-IR than
eggs was accompanied by a rise in lactic and succinic acids. with chemical methods. Chong et al. (20M) applied 14-MeV
Mollica et al. (39L) determined the nonvolatile organic acids neutron activation analysis to measure organic nitrogen by
in sugar maple sap by TMS-GC on a mixed liquid phase utilizing carbon and oxygen interference. An automated
column of 4% SE-52/2% SE-30 on Chromsorb W-HP with proton activation analysis technique was described by Con-
N as carrier gas and FID detection. stantinescu (22M) to measure total nitrogen levels in cereals.
Oen et al. (41L) analyzed sweet cherry cultivars for L-malic, Overley (79M) measured the nitrogen, hydrogen, carbon, and
quinic, citric, chlorogenic, phosphoric, galacturonic, glucuronic, oxygen contents of cereals and other chemical compounds
glucaric monolactone, gluconic, and oxalic acids by precipi- using neutron attenuation analysis. Protein in cereal meals
tation of the acids as lead salts, liberating the free acids, was also measured with 14-MeV neutron activation analysis
drying, and analyzing with TMS-GC and GC/MS. Butyric by Samei et al. (90M).
and caproic acids were determined in milk fat by reaction with A monospecific antiserum was developed by Barnett and
methanolic barium hydroxide, filtering, releasing the two acids, Howden (7M) using a heat-resistant glycoprotein antigen from
and GC analysis on Chromosorb W AW 100/120 with 15% peanuts and was applied to the detection of heat treatment
SP-1220/1% phosphoric acid as reported by Spahis et al. via rocket immunoelectrophoretic techniques. Two sandwich
(52L). Tsuda et al. (53L) analyzed soft drinks and jams for ELISA methods having little cross reactivity were developed
their content of sorbic, dehydroacetic, benzoic, malic, and by Fritschy et al. (34M) to detect both wheat a-gliadins and
tartaric acids by ion exchange of extracts and liquid-liquid whole gliadins. McKillop et al. (70M) also developed an
partition of acids. Sorbic, dehydroacetic, and benzoic acids ELISA test for wheat gliadins. Enzyme-coupled monoclonal
were gas chromatographed on a 5% DEGS +1% phosphoric antibody tests for different wheat proteins, including gluten,
acid column whereas succinic, fumaric, malic, and tartaric gliadin, and cooked gluten, were developed by Skerritt et al.
acids were TMS-GC determined on a 3% SE-30 column. (94M), Skerrit (95M), and Skerritt and Smith (96M). Greiner

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 .235R
FOOD

et al. (39M) developed an immunoturbidimetric assay for the differences using polarographic analysis. Recovery of me-
detection of whey protein in nonfat dry milk using antibodies thionine from feed concentrates was improved by adding water
raised against whole bovine whey. An ELISA test for quan- to the formic acid/hydrogen peroxide oxidation mixture ac-
titative determination of denatured /3-lactoglobulin in milk cording to Slump & Bros (97M). Williams et al. (108M)
was described by Heppell (45M). Protein lactosylation was measured the methionine content of peas. Eka and Oyeleke
detected by Matsuda et al. (68M) using antibodies specific (26M) preferred pronase enzyme digestion of sorghum grain
for <-deoxylactulosyl lysine. Immunochemical methods for and maize over acid or base hydrolysis for tryptophan de-
the measurement of a variety of food ingredient proteins were terminations. In a comparison of ion exchange HPLC with
presented by Menger et al. (71M). An ELISA method for the conventional ion exchange chromatographic amino acid de-
detection of soy protein in soy lecithin, soy oil, and soy-based terminations, Elkin and Griffith (26M) found that HPLC is
margarine was reported by Porras et al. (85M). more reproducible with respect to retention times but yields
Andrews et al. (3M) utilized fast protein ion exchange liquid lower tyrosine and phenylalanine values and is less reprodu-
chromatography with Mono Q and Mono S resins to resolve cible with respect to peak area. Elkin (30M) also compared
major milk protein classes; a si, a s2,3-, *-, and 7-caseins were reversed-phase HPLC of OPA-amino acids with conventional
resolved on Mono S, and whey proteins were resolved on Mono ion exchange chromatographic techniques and found that both
Q. Both casein and whey proteins were separated by Bican methods yield comparable results; however, proline and
(14M) using a Protein Pak DEAE-5PW ion exchange HPLC cysteine were not detected by HPLC and glycine and threo-
column. Carpenter and Brown (18M) applied size exclusion nine were not completely resolved. Friedman and Noma
HPLC to resolve and quantitate casein micelles after treat- (33M) reported the analysis of (phenylethyl)aminoalanine and
ment with calcium. Gel filtration on Sephacryl S-1000 sep- lysinoalanine in casein and soy protein using ion exchange
arated casein micelles by size and from whey according to chromatography. Gehrke et al. (36M) recommended higher
Ekstrand and Larsson-Raznikiewicz (27M). Green (38M) acid hydrolysis temperatures (145 °C) for shorter durations
resolved x-casein by polyacrylamide gel electrophoresis and (4 h) to achieve greater precision from subsequent chroma-
characterized the fractions by chymosin sensitivity and by tography of liberated amino acids. Huet and Pernollet (48M)
staining with Ethyl-Stains-all. Manji et al. (66M) fractionated determined tryptophan after barium hydroxide digestion by
whey proteins by anion exchange chromatography eluting with chromatography on Fractogel TSK HW 40 S and postcolumn
a sodium acetate ionic strength gradient. Two-dimensional derivitization with o-phthalaldehyde. A gas chromatography
PAGE analysis of wheat proteins, incorporating isoelectric method for enzyme-released amino acids, as N-trifluoroacetyl
focusing and SDS-PAGE, resolved 480 discrete spots according butyl esters, was reported by Ihekorouye (50M). Ingles and
to Lei and Reeck (60M). Lei and Reeck (61M) also compared Gallimore (52M) described a reversed-phase HPLC method
two-dimensional gels of triticale proteins with those from their for the separation of amino acids derivitized with fluoroes-
parental durum and rye species. A PAGE system for wheat camine. Kuninori and Nishiyama (59M) separated and
gliadins was optimized by Lookhart et al. (63M) with respect quantitated free and bound ferrulic acid as well as tyrosine
to operating temperature and buffer composition. Wheat in wheat seeds using reversed-phase HPLC and postcolumn
gliadins were fractionated by Popineau and Pineau (84M) fluorescence spectre! analysis. Reaction of «-trinitrophenyl-
using SP Trisacryl M, a mechanically and chemically stabilized lysine with carbohydrates during hydrolysis in the trinitro-
cation exchange resin. benzenesulfonate method was reduced by James and Ryley
Cuq and Cheftel (24M) reviewed detection methods, bio- (53M) by minimizing hydrolysis times. Molnar-Perl et al.
logical effects, and toxicity of lysinoalanine formed by thermal (73M) applied several dye-binding techniques, including
and alkaline processes. The detection of biologically available Orange G, acid orange 12, and amido black 10B, to the de-
lysine via chemical, biological, microbiological, and enzymatic termination of reactive lysine. Perl et al. (83M) determined
methods was reviewed by Zomborszky (110M). Beutler (12M) reactive lysine by selective removal of reactive residues by
described an enzymatic method for glutamate determination modification with l-phenylazo-2-naphthol-6,8-disulfonic acid
where formazan, resulting from the combined actions of glu- and acylation with propionic anhydride (histidine and arginine
tamate dehydrogenase and diaphorase with NAD/NADH and also react with this reagent). Ray (87M) measured available
INT, is measured at 492 nm. Beveridge and Harrison (13M) lysine by reacting the intact protein with 2,4-dinitrofluoro-
modified the 2,4,6-trinitrobenzenesulfonate method for amino benzene followed by hydrolysis and thin-layer chromato-
nitrogen in juice, juice concentrate, and liquid fruit products. graphic resolution. Tomarelli et al. (98M) determined reactive
Lysinoalanine was measured as its heptafluorobutyryl de- lysine in infant formulas by reaction with trinitrobenzene-
rivative by Bueser and Erbersdobler (15M) using GLC and sulfonate, hydrolysis, and reversed-phase HPLC analysis with
a 3% SE-30 column. Precolumn derivitization of proline and detection at 346 nm. A microbiological method for nutri-
hydroxyproline with 4-chloro-7-nitrobenzofurazan afforded tionally available lysine was developed by Tuffnell and Payne
Carisano (17M) a sensitive fluorometric HPLC assay. Chung (99M) using an auxotroph of E. coli which synthesizes /3-ga-
et al. (21M) assessed amino acid racemization in alkalie-treated lactosidase enzyme in proportional response to lysine; the
proteins via a dextrorotatory-specific enzyme reactor composed galactosidase activity was measured by conventional proce-
of D-amino acid oxidase and catalase immobilized on porous dures. A rapid procedure for determination of lysine was
succinamidopropyl glass beads. Csiba (23M) developed a described by Viroben (104M) where total lysine is measured
colorimetric method for hydroxyproline that is amenable to by reaction ith Acid Orange 12 before and after sample re-
automation and depends on the reaction with p-(dimethyl- action with propionic anhydride. Picomole detection levels
amino)benzaldehyde. In a study on sulfur-containing amino for OPA-derivitized amino acids resolved by HPLC were
acid recovery from proteins, Dennison and Gons (25M) de- achieved by Lookhart and Jones (62M) by using ethanethiol
termined that recoveries vary with the food matrix regardless rather than mercaptoethanol during derivitization. An en-
of oxidation and hydrolysis conditions employed. By oxidizing zymatic method to determine aspartic acid (and asparagine
sorghum with performic acid prior to HC1 hydrolysis, Elkin after deamination) was outlined by Moellering (72M) where
and Griffith (28M) observed increases in cysteine, methionine, aspartic acid is converted to oxaloacetic acid which is reduced
and histidine recoveries and destruction of tyrosine and by NADH in the presence of malate dehydrogenase; NADH
phenylalanine; these authors suggested that one sample consumption is followed at 339 nm. Ribarova and Shishkov
preparation method is not suitable. Gustine (42M) measured (88M) reported an improved equation to calculate the amino
S-methylcysteine sulfoxide in Brassica extracts by reversed- acid contents of foodstuffs from hydrolysis-chromatography
phase HPLC with fluorescence detection after precolumn data. Nielsen and Hurrell (76M) determined tryptophan by
derivitization with o-phthalaldehyde. vacuum hydrolysis with lithium or sodium hydroxide followed
Methionine, ass-
methionine sulfoxide, was determined by Hayashi and Suzuki by reversed-phase HPLC with fluorescence detection. Sau-
(44M) in proteins by hydrolyzing with 3 N p-toluenesulfonic ra-Calixto and Canellas Lourdes Soler (91M) determined
acid at 110 °C for 22 h. MacDonald et al. (64M) reported on tryptophan in nuts and seeds by ion exchange chromatography
a collaborative study where sulfur-containing amino acids were and postcolumn reaction with p-(diethylamino)benzaldehyde
measured in food and feed proteins by ion exchange chro- and glyoxylic acid followed by spectrophotometric analysis.
matography following oxidation with performic acid prior to Tyrosine and histidine were determined as red azo dyes by
HC1 hydrolysis; the method was adopted as official first action. Tummuru and Sastry (100M) following reaction with diazo-
In a study on the inter- and intramolecular cystine contents tized 4-chloro-2-nitroaniline. Ukeda et al. (101M) reported
of wheat gliadin proteins, Nadirov (74M) found interbiotype a rapid protein measurement for dairy products whereby the

236R* ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
 FOOD

reaction of glutaraldehyde with protein is measured amper- change HPLC with electrochemical detection. In a study on
ometrically using a Clark oxygen electrode. When measuring biogenic amines in cheese, Zee et al. (109M) found tri-
3-methyl-L-histidine in muscle tissue, White and Lawrie chloroactic acid to be the best extraction solvent, but ex-
(107M) found that recoveries from acetone powders were traction efficiencies varied with cheese type, amine species,
superior to those from fresh tissue. and relative concentration of the amine.
Arnstadt (5M) measured urea in urease-treated milk by
reaction with p-(dimethylamino)benzaldehyde and colorimetry VITAMINS
at 435 nm. Beattie et al. (8M) utilized HPLC-MS with a Books that are of use to the vitamin analyst for review and
moving belt interface and ammonium chemical ionization to reference are Methods of Vitamin Assay—Fourth Edition
measure nonvolatile nitrosamines; silylation of the polyimide (4N), Methods for the Determination of Vitamins in Foods
belt helped to reduce decarboxylations and denitrosations that (ION), and Modern Chromatographic Analysis of Vitamins
occurred during transfer. Gavinelli et al. (35M) determined (15N). The separation of fat-soluble vitamins by HPLC from
seven volatile nitrosamines by simultaneous distillation-ex- fish products was discussed by Lambertsen (36N). Methods
traction and GC analysis with thermal energy detection. Sen for vitamins A, C, D3, and E in dietetic foods by HPLC were
et al. (92M) minimized artifact nitrosamine formation from reported by Sell (49N). Gas chromatographic methods for
the analysis of rubber baby bottle nipples by pretesting fat-soluble vitamins in foods were reviewed by Davidek et al.
methylene chloride for nitrosation potential and by incorpo- (14N) and water-soluble vitamins were reviewed by Velisek
rating propyl gallate (an inhibitor of nitrosation). An enzy- et al. (59N).
matic method for ammonia was reported by Bergmeyer (11M) Bognar (8N) described an HPLC method for vitamin A in
whereby NH3 is consumed in the conversion of 2-oxoglutarate foods and reported the results of a collaborative study. The
to glutamic acid with simultaneous loss of NADH. A con- Carr-Price and an HPLC method for vitamin A in fortified
tinuous flow apparatus for ammonia and urea (as ammonia) milk were compared by Mills (42N) and found to be statis-
measurement was described by Kirst et al. (57M) where upon tically equivalent. Coverly (13N) modified a continuous flow
reaction with perchlorate in the presence of nitroprussate, a method for the determination of vitamin A in milk products
blue colored reaction product is formed. A flow injection and infant formula to increase the precision, recovery, and
analysis for urea in cow’s milk was reported by Oltner et al. sampling rate. Speek et al. (53N) described a method for the
(77M) where an enzyme reactor containing urease generates determination of /3-carotene by HPLC and the calculation of
ammonia gas which is separated by a gas diffusion cell and vitamin A activity in vegetables. Stancher et al. (54N) de-
measured by bubbling it into a color reaction solution. Hy- termined the isomer distribution of retinol and 3-dehydro-
poxanthine in fish was determined by Burns and Ke (16M) retinol by HPLC and determined vitamin A active compounds
using reversed-phase HPLC with a mobile phase consisting in fish oils. A TLC procedure for the determination of vitamin
of 0.01 M potassium phosphate, pH 4.5. Grassl and Supp A in foods was suggested by Gerstenberg (22N) using
(37M) described an enzymatic determination for inosine fluorescence measurement after saponification and extraction.
monophosphate whereby IMP is converted to hypoxanthine A reversed-phase HPLC method for the determination of
and ribose-5-phosphate and the hypoxanthine is oxidized to vitamin A palmitate in nonfat milk and vitamin D in whole
uric acid which is measured at 293 nm. The effects of pH, milk was described by Grace et al. (23N). HPLC methods
temperature, and ionic strength on ion-pair reversed-phase for vitamin D in foods were reviewed by Kobayashi et al. (34N)
HPLC analysis of purine nucleotide monophosphates were with emphasis on a method developed by the authors in 1981.
studied by Mack et al. (65M). Nguyen et al. (75M) determined Vitamin D3 in fat, oil, and margarine was determined by
ribonucleoside 5'-monophosphates and their isomers in potatoe Rychener et al. (46N) by HPLC using vitamin D2 as an in-
tubers simultaneously by ion exchange HPLC. Ryder (89M) ternal standard. Vitamins D2 and D3 were determined by
described ion exchange HPLC methods for the measurement liquid chromatography in fortified milk and infant formulas
of ATP and breakdown products in perchloric acid extracts by Landen (37N) after fractionating the vitamins from the
of muscle tissue. Chang et al. (19M) determined histamine, lipid material by using gel permeation chromatography. Sertl
tyramine, tryptamine, histidine, tyrosine, and tryptophan in et al. (50N) also determined vitamins D2 and D3 in milk and
cheese using reversed-phase HPLC with a counterion-con- infant formulas by normal-phase LC after using an amino-
taining mobile phase. The agaritine content of commercial cyano LC cleanup column to remove major interferences.
canned mushrooms was measured by Fischer et al. (32M) using HPLC procedures for tocopherols were described by Shen et
cation exchange HPLC on Partisil SCX at pH 1.8. An IUPAC al. (51N) who separated a-, ft-, y-, and 5-tocoherols on a short
recommended method for determining azaarenes in meat using column to reduce analysis time, by Speek et al. (52N) who
capillary GC profiling was reported by Grimmer and Naujack determined the four tocopherols, and a-tocotrienol in oil seeds
(40M). Grivas and Nyhammer (41M) compared electro- using electron-impact mass spectroscopy for standardization,
chemical methods with UV methods for quantitation of re- and by Rammell et al. (45N) who separated the four toco-
versed-phase HPLC separated mutagenic imidazoquinolines pherols and their corresponding tocomononals, and tocotrinols,
and imidazoquinoxalines. Ingles et al. (51M) detected biogenic and plastochromanol-8 in six seed oils using an amino-cyano
amines in foods by ion exchange cleanup and HPLC analysis HPLC column. A routine method for the determination of
of fluorescent derivatives; positive identification was achieved a-tocopherol acetate in fortified milk powder formulation
by field desorption MS. a GC-FID method for the analysis using HPLC was described by Woollard et al. (63N). Vitamin
of allylisothiocyanate and /3-phenylethylisothiocyanate was Kj in infant formulas was determined by Hwang (29N) using
described by Kojima et al. (58M). Long chain aliphatic amines HPLC and UV detection. Zonta et al. (64N) determined
in fats were detected by Matschiner et al. (67M) upon reaction vitamin Kx in soy bean oil after enzymic digestion, extraction,
with 4-methyoxy-7-nitrobenz-2,l,3-oxadiazole and separation and alumina-column cleanup, followed by isocratic reversed-
by TLC. Mayanna and Jayaram (69M) developed a method phase chromatography with UV detection.
for dissolved caffeine by oxidimetric titration with aromatic Thiamin in foods was determined by Velisek et al. (58N)
sulfonyl A’-haloamines. Patterson (81M) described a new by gas chromatography with a flame photometric detector.
flame thermionic ionization detector for the Iatroscan rod Echols et al. (16N) evaluated solvents and recommended
chromatography system that is specific for nitrogen- and internal standards for the gas chromatographic determination
halogen-containing compounds. Peleran and Bories (82M) of thiamin. Free and total thiamin in milk were determined
measured indole and 3-methylindole in pig fat back by com- by gas chromatography by Echols et al. (17N) Using a nitro-
bining selective solvent extraction with GC resolution and gen-phosphorus detector. Thiamin in infant formula was
N/P-sensitive detection; identities were confirmed by MS. A determined by Ayi et al. (5N) by liquid chromatography using
multienzyme sequence to determine creatine and creatinine UV detection. Kneifel et al. (31N) determined thiamin in milk
was reported by Wahlefeld and Siedel (105M) where creatine products by HPLC after oxidation to thiochrome and sub-
is phosphorylated yielding ADP, the liberated ADP and sequent fluorescence detection. Recent methods for the de-
phosphoenolpyruvate are converted to ATP and pyruvate, and termination of thiamin and riboflavin are discussed and a new
the pyruvate is converted to lactate with simultaneous loss method for the simultaneous determination of these two vi-
of NADH. Wakabayashi (106M) developed a scheme to tamins is proposed by Augustin (3N). The use of HPLC for
measure heterocyclic amines in cooked foods which incorpo- the simultaneous determination of thiamin and riboflavin was
rates treatment with blue cotton, partitioning against 0.1 N described by Finglas et al. (20N) in potatoes, by Mauro et al.
HC1, silica gel cleanup, and either reversed-phase or ion ex- (41N) in enriched cereal products, and by Wimalasiri et al.

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 237 R
FOOD

C62N) in foods using both UV and fluorescence detection. third in a series devoted to a discussion of recent developments
Wills et al. (61N) determined thiamin and riboflavin in foods in food analysis procedures, Analysis of Foods and Beverages,
by HPLC and fluorometric methods and preferred the HPLC Modern Techniques (4P), and The Use of Statistics to De-
method for speed and accuracy. velop and Evaluate Analytical Methods (21P).
The use of HPLC for the determination of riboflavin was A review discussion the application of near-infrared spec-
described by Ashoor et al. (2N) in milk and dairy products troscopy (near-IR) to the analysis of food and feed with em-
with UV detection, by Kneifel et al. (33N) in milk by phasis on the measurement of moisture, protein, oil, starch,
fluorescence detection, and by Watado et al. (60N) in fresh and fiber was published by Bernardini et al. (2P). Ronalds
fruits and vegetables with ion-pairing chromatography and and Miskelly (17P) also reviewed near-IR applications in food
fluorescence detection. On-line enrichment and isolation of analysis. Frankhuizen and Van der Veen (7P) reported on
vitamin B2 from large volumes of aqueous food samples fol- the calibration of near-IR reflectance apparatus for the rapid
lowed by ion-pairing chromatography was described by Jau- determination of major and minor constituents in milk powder
mann et al. (30N). Riboflavin was determined by Colugnati and cheese. Near-IR was investigated by Sato et al. (18P) for
(12N) in fortified powdered milk and infant formula using measuring major constituents of raw milk and found the
cyclic pulsed voltammetry. An ion-pair liquid chromato- technique useful for measuring fat, protein, lactose, and total
graphic method for the simultaneous determination of vita- milk solids. Nadai and Mihalyi-Kengyel (14P) investigated
mins B2 and B6 in infant formula products was proposed by equations for predicting moisture, fat, and protein of raw meat
Ayi et al. (6N). by near-IR and Valdes and Summers (20P) investigated this
A microbioassay procedure for the determination of free technique for the determination of protein and fat in raw
and bound niacin in cereals was described by Aki et al. (IN). poultry. The rapid analysis of food packaging laminates by
On-line generation of cyanogen chloride in the semiautomated near-IR was reported by Davies et al. (6P).
determination of niacin and niacinamide in food products was A review article was published by Giacin and Brzozowska
proposed by Ge et al. (21N). An automated photometric (8P) on the migration of substances from food-contacting
method for the determination of niacin and niacinamide in materials into foods and methods for determining such sub-
grain and cereal products was proposed by Holz (28N) using stances. Hischenhuber (9P) reviewed the use of HPLC for
1,3-dimethylbarbituric acid and on-line generation of cyanogen determining food components, additives, and contaminants.
chloride. A reversed-phase ion-pair HPLC method for the A review of the application of HPLC in dairy science was
determination of nicotinic acid in instant coffee was described reported by Miebs and Kirst (12P) including dairy product
by Trugo et al. (55N). components, mycotoxins, contaminants, impurities, and
The use of HPLC for the determination of pyridoxin and preservatives. Woollard (22P) reviewed the expanding use
the several forms of the vitamin in foods was described by of HPLC in the analysis of food for proteins, fats, carbohy-
Bognar et al. (8AO, by Vanderslice et al. (56N), and by Gregory drates, trace elements, amino acids, carboxylic acids, vitamins,
et al. (25N). A cation-exchange HPLC method for the seven additives, antioxidants, preservatives, coloring and flavoring
major metabolic forms of vitamin B6 was described by Cobum materials, contaminants, aflatoxins, and other constituents.
et al. (11N). Ekanayake et al. (18N) described an in vitro, The application of ion chromatography in the food and bev-
two-stage enzymic digestion system followed by HPLC to erage industry was reviewed by Cox et al. (5P); sample
determine biologically available vitamin B6 in vitamin Be preparation and apparatus are covered in the article.
fortified foods. Oesterdahl et al. (43N) compared a radioi- Apparatus and methods for analysis of foods by differential
sotope dilution (RID) method for the determination of vitamin thermal analyses and calorimetry were described by Raemy
B12 in cereal with a standard microbiological assay with et al. (16P) who discussed thermal properties of carbohydrates,
Lactobacillus leichmannii and found the RID method ac- lipids, and proteins. Instruments and sensors for carrying out
ceptable for routine determination of vitamin B12. Gregory chemical, biochemical, and immunological methods of food
(24N) reviewed liquid chromatographic methods for deter- analyses were reviewed by Kress-Rogers (IIP). A review on
mining folacin in food and other biological materials. Folic immunoassays in food analysis including principles of ELISA,
acid in commercial diets was determined by Schieffer et al. development of kits, and applications was published by Allen
(47N) using anion-exchange solid-phase extraction and sub- (IP). Test strips were compared by Schwedt and Altkofer
sequent reversed-phase HPLC. The determination of the (19P) with standard analytical procedures in the semiquan-
biotin content in foods by means of a protein binding assay titative determination of pH, K, nitrate, sulfite, tartaric acid,
was reported by Bitsch et al. (7N). A radiometric microbio- and ascorbic acid in several foods and alcoholic beverages;
logical assay using Kloeckera brevis was used by Guilarte results compared well although in some cases a simple cleanup
(26N) for the analysis of biotin in a variety of food samples. procedure was necessary. Methods for the surveillance of
Analytical methods for determining ascorbic acid in food radioactivity in food originating from natural and artificial
products, biological samples, and pharmaceuticals were re- sources were described by Boppel et al. (3P). A method to
viewed by Pachla et al. (44N) covering spectroscopic, elec- determine radionuclide levels in food by rapid pretreatment
trochemical, enzymic, and chromatographic techniques. Both of samples with a modified microwave dehydration apparatus
L-ascorbic and L-dehydroascorbic acid were determined by followed by radiation counting was developed by Nakaoka et
Kneifel et al. (32N) in milk and whey by paired-ion re- al. (15P).
versed-phase HPLC. An HPLC method was used by Kodaka
et al. (35N) for the determination of total vitamin C after ACKNOWLEDGMENT
precolumn derivatization with 2,4-dinitrophenylhydrazine. The authors gratefully acknowledge the efforts of Carol
Seki et al. (48AO separated ascorbic acid, dehydroascorbic acid,
and diketoglulonic acid by HPLC with fluorometric detection Butler, Allen Oliver, and Nancy Ernst for computer assistance
and applied the method to the determination of ascorbic acid and Miriam Verner for typing the manuscript.
in fruit juice. A variety of foods were analyzed for vitamin LITERATURE CITED
C by Vanderslice et al. (57N) by HPLC and fluorometric
detection. Photometric flow injection methods for the de- ADDITIVES
termination of ascorbic acid were described by Hernandez- (1A) Ali, M. Sher, J. Assoc. Off. Anal. Chem., 1985, 68, 488. CA103-
Mendez et al. (27N) and by Lazaro et al. (40N). A UV method (1):5100n.
for the determination of ascorbic acid in soft drinks, fruit (2A) Argoudelis, Chris J., J. Chromatogr., 1984, 303, 256. CA101-
(25):228621t.
juices, and cordials was described by Lau et al. (38N). Dif- (3A) Arneth, Wolfgang; Witzgall, Adelheid, Fleischwirtschaft, 1985, 65, 637.
ferential pulsed polarography was used by Lau et al. (39N) CA103(9):69832k.
for the determination of ascorbic acid in vegetables and fruits. (4A) Barnett, Don, Food Technol. Aust., 1985, 37, 503. CA704(1):4680k.
Esaka et al. (19N) determined L-ascorbic acid in foods with (5A) Bauer, Friedrich; Stachelberger, Herbert, Chem., Mikrobiol., Technol.
immobilized ascorbate oxidase. Lebensm., 1984, 8, 129. CA101( 15):128969u.
(6A) Beutler, H. 0., Food Chem., 1984, 15, 157. CA 707(25):228591h.
(7A) Blanchflower, W. John; Rice, D. A.; Hamilton, J., Analyst (London),
MISCELLANEOUS 1985, 110, 1283. CA104( 17):147252h.
Books that are of use to the food analyst for review and (8A) Brumley, W. C.; Warner, C. R.; Daniels, D. H.; Andrzejewski, D.; White,
K.; Min, Z.; Cen, J.; Sphon, J., J. Agric. Food Chem., 1985, 33, 368.
reference are: Immunoassay in Food Analysis (13P), which CA 702(23):202671e.
covers principles of immunoassay and applications, Devel- (9A) Chester, T. L.; Innis, D. P.; Owens, G. D., Anal. Chem., 1985, 57,
opments in Food Analysis Techniques—3 (10P), which is the 2243. CA 103(15):121827d.

238 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
 FOOD

(10A) Chlkamoto, Takejl; Maitani, Takeshi, Shokuhin Elselgaku Zasshi, 1984, (56A) Yanai, Kiyoshi, Japan, 1985, CA 104(3):18867p.
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ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 239 R
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Sci., 1985, 36, 263. CA105( 15):132217x. (109B) Hatanaka, Hisakatsu; Kaneda, Yoshio, Shokuhin Eiseigaku Zasshi,
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244 R. ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
 FOOD

(68F) Hibino, Hidehiko; Fukudo, Nobuo; Kudo, Kiyoshi, Yukagaku, 1985, 34, (116F) Martovshchuk, V. I.; Mgebrishvili, T. F.; Martovshchuk, E., Maslo
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ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 245 R
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'

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(100J) Van Betteray-Kortekaas, A. M. G.; Vos, G., Fresenius’ Z. Anal. 649. CA 702(9):75031u.
Chem., 1986, 323, 493. CA104(23):205639c. (21L) Joex, E.; Pietzsch, W.; Froede, M., Lebensmittelindustrie, 1986, 33,
(101J) Van Eeden, C. H. Peter; De Jong, Alex W. J., Z. Lebensm. Forsch., 79. CA 705(7):59525q.
1985, 181, 412. CA 104 (3): 18704h. (22L) Kataglrl, Mltsuaki; Shimizu, Sumio; Kaihara, Hiromichi, Nippon Nogel
(102J) Viladrlch Gonzalbez, E.; Forcadell Berenguer, M.; Buxaderas San- Kagaku Kaishi, 1986, 60, 385. CA705(11):96108b.
chez, S.; Merine-Font, A., Grasa Aceltes, (Seville), 1986, 37, 77. (23L) Kayahara, Hisataka; Yasuhara, Hitomi, Miso no Kagaku to Gijutsu,
CA 105(9):77645h. 1984, 32, 242. CA707(15): 128962m.
(103J) Vlacil, F.; Vins, I., Nahrung, 1985, 29, 467. CA703(11):86580u. (24L) Kikunaga, Shigeshl; Takahashi, Masayuki, Nippon Eiyo, Shokuryo Gak-
(104J) Warner, C. R.; Daniels, D, H.; Joe, F. L.; Fazio, T., J. Assoc. Off. kaishi, 1985, 38, 123. CA703(17): 140393a.
Anal. Chem., 1986, 69, 3. CA104( 11):87228t. (25L) Kogure, Kyolchi; Enomoto, Yasunori; Shibata, H., Patent, 1985.
(105J) White, R. Thomas, Jr.; Douthit, Garnett E., J. Assoc. Off. Anal. CA 704(13): 108203y.
Chem., 1985, 68, 766. CA103( 13):101265r. (26L) Kok, W. T.; Groenendijk, G.; Brinkman, U. A. T.; Frei, R. W., J. Chro-
(106J) Wichman, M. D.; Fietkau, R.; Fry, R. C., Appl. Spectrosc., 1986, 40, matogr., 1984, 315, 271. CA 702(9):75060c.
233. CA104( 15):128359g. (27L) Lagemann, M.; Graef, V.; Anders, D., Dtsch. Lebensm .-Rundsch,
(107J) Wlechen, A.; Kock, B., Fresenius’ Z. Anal. Chem., 1984, 319, 569. 1985,67,140. CA703(13):1O3523x.
CA 702(5):44475n. (28L) Lamkin, W. M.; Luginsland, N. D.; Pomeranz, Y., Cereal Chem., 1985,
(108J) Yan, Daren; Stumpp, E.; Schwedt, G., Fredsenius’ Z. Anal. Chem., 62,6. CA 702(21):183829u.
1985,322,474. CA704(5):33137u. (29L) Lamkin, W. M.; Unruh, N. C.; Pomeranz, Y., Cereal Chem., 1986, 63,
(109J) Yan, Daren; Schwedt, Georg, Fresenius' Z. Anal. Chem., 1985, 372. CA 705(21):189558p.
320, 252. CA102(16):142487n. (30L) Lin, L.; Tanner, H., HRC CC, J. High Resolut. Chromatogr. Chroma-
MOISTURE togr. Commun., 1985, 6, 126. CA702( 19): 165344c.
(31L) Littmann, S., Dtsch. Lebensm. 1985, 81, 345. CA704(5):33146w.
(IK) Bussiere, G.; Serpelloni, M., NATO ASI Ser., 1985, Ser. E, 90(Prop. (32L) Malmberg, Alf G.; Theander, Olof, J. Agric. Food Chem., 1985, 33,
Water Foods), 627. CA 703(15):121826c. 549. CA 702(23):202686p.
(2K) Chin, H. B.; Kimball, J. R.; Hung, J.; Allen, B., J. Assoc. Off. Anal. (33L) Matsumoto, K.; Ishida, K.; Nomura, T.; Osajima, Y., Agric. Biol.
Chem., 1985, 68(6), 1081-3. Chem., 1984, 48, 2211. CA707(23):209213v.
(3K) Chirife, Jorge; Resnlk, Silvia L., J. Food Sci., 1984, 49, 1486. CA707- (34L) Matsuo, T.; Yorimitsu, A.; Kanehiro, M.; Kanamori, H.; Sakamoto, I.,
(25):228633y. Hiroshima ElseKenkyusho Kenkyu Hokoku, 1984, 37, 47. CA103-
(4K) Gambhir, P. N.; Agarwala, A. K., JAOCS, J. Am. Oil Chem. Soc., (23):195038u.
1985, 62, 103. CA 102(9):77350w. (35L) Mattluz, Edward L.; Perone, Ruth A., LC, 1986, 4, 552. CA105-
(5K) Gerschenson, Lia; Favetto, Guillermo; Chirife, J., Lebensm.-Wiss. (5):41355z.
Technol., 1984, 77,342. CA 702(9):77352y. (36L) Mazzola, E. P.; Phillippy, B. Q.; Harland, B.; Miller, T.; Potemra, J.;
(6K) Kitic, D.; Pereira Jardlm, D. C.; Favetto, G.; Resnick, S.; Chirife, J., J. Katsimpiris, E., J. Agric. Food Chem., 1986, 34, 60. CA104-
Food Sci., 1986, 57, 1037. CA705(13):113772h. (11 ):87191a.
(7K) Koizumi, Hideo; Yasui, Akemi; Tsutsumi, Chiuchi, Nippon Shokuhin Ko- (37L) Mentastl, E.; Gennaro, M. C.; Sarzanini, C.; Baiocchi, C.; Savigliano,
gyo Gakkalshi, 1984, 37, 465. CA 707(21): 189822n. M„ J. Chromatogr., 1985, 322, 177. CA702(21):183689y.
(8K) Labuza, T. P., Moisture Sorption: Practical Aspects of Isotherm Mea- (38L) Moellerlng, Hans, Methods Enzym. Anal., 1985, 7, 2. CA103-
surement and Use: Am. Assoc, of Cereal Chem. (AACC): St. Paul, MN, (21):174758p.
1984. (39L) Mollica, Joseph N.; Morselli, Maria Franca, J. Assoc. Off. Anal.
(9K) Rueegg, M.; Steiger, G.; Moor, Ursula, Mitt. Geb. Lebensmlttelunters. Chem., 1984, 67, 1125. CA702(13):111585p.
Hyg., 1986, 77, 139. CA705(31):113716t. (40L) Murakami, R.; Yamamoto, K.; Kamiya, T.; Komuro, M.; Kakefuda, S.;
(10K) Rueegg, M.; Moor, Ursula, Mitt. Geb. Lebensmlttelunters. Hyg., 1986, Takai, K., Shokuhin Elseigaku Zasshi, 1984, 25, 360. CA101-
77, 131. CA705(13):113715s. (25):228616v.
(11K) Sanna, Louis A., J. Assoc. Off. Anal. Chem., 1986, 69, 834. (41L) Oen, Haakon; Vestrheim, Sigbjoern, Acta Agric. Scand., 1985, 35,
CA 705(21):189577u. 145. CA 703(19): 159329f.
(12K) Smith, D. M.; Mitchell, J., Aquametry Part 2. Electrical and Electronic (42L) Ohkawa, Hironorl, J. Assoc. Off. Anal. Chem., 1985, 68, 108.
Methods: Wiley: Chicester, Sussex, 1984. CA 702(21):183841s.
(13K) Stamp, J. A.; Linscott, S.; Lomauro, C.; Labuza, T. P., J. Food Scl., (43L) Otsuka, K.; Horl, M.; Sugitani, A.; Yamada, F., GIFU EISEI KENKYU-
49, 1139. CA 707(19): 189202g. SHOHO, 1985, 30, 67. CA 705(17): 151616T.
(14K) Ueda, Masafumi; Shiba, Kamekichl, Moisture Humidity, Proc. Int. (44L) Panarl, G., Mllchwissenschaft, 1986, 47, 214. CA 705(1):5246f.
Symp., 1985, 751. ISA: Research Triangle Park, NC. CA104- (45L) Picha, David H., J. Agric. Food Chem., 1985, 33, 743. CA103-
(4):27977b. (5):36256x.
ORGANIC ACIDS (46L) Proohazka, Lubos; Kvasnicka, Frantisek; Stechova, Alena, Kvasny
Prum., 1986, 32, 33. CA705(7):59529u.
(IL) All, M. Sher, J. Assoc. Off. Anal. Chem., 1985, 68, 488. CA103- (47L) Puttemans, Marc L.; Dryon, Louis; Massart, Desire, J. Assoc. Off.
(1):5100n. Anal. Chem., 1984, 67, 880. CA707(23):209199v.

ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987 • 249 R
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(48L) Puttemans, Marc L.; Branders, Chris; Dryon, Louis; Massart, Desire, J. (42M) Gustine, David L., J. Chromatogr., 1985, 379 , 450. CA102-
Assoc. Off. Anal. Chem., 1985, 68, 80. CA 102(21):183839x. (15): 130537r.
(49L) Rabe, E., Veroeff. Arbeltsgem. Getreideforsch., 1984, 35, 114. (43M) Hach, Clifford, C.; Brayton, Scott V.; Kopelove, A., J. Agric. Food
CA 703(17):140403d. Chem., 1985, 33, 1117. CA703(25):210255h.
(50L) Rocklin, Roy D.; Slingsby, Rosanne, W.; Pohl, C. A., J. Uq. Chroma- (44M) Hayashl, Rlklmaru; Suzuki, Fukuko, Anal. Biochem., 1985, 149, 521.
togr., 1986, 9, 757. CA104(23):203300m. CA 703(17): 14041 Od.
(51L) Schaller, Karl Heinz; Triebig, Gerhard, Methods Enzym. Anal., 1984, (45M) Heppell, L. M. J., Immunoassays Food Anal., 1985, 521. CA103-
6, 668. CA 702(9):75O32v. (19): 159161 b.
(52L) Spahis, Khira; Kuzdzal-Savoie, Simone, Lait, 1984, 64, 579. CA103- (46M) Honlgs, D. E.; Hieftje, Gary M.; Mark, H. L.; Hlrschfeld, T. B., Anal.
(15): 121852h. Chem., 1985, 57, 2299. CA703(15):121814x.
(53L) Tsuda, T.; Nakanishi, H.; Morita, T.; Takebayashi, J., J. Assoc. Off. (47M) Horvath, L.; Norris, K.; Horvath-Mosonyi, M., Acfa Aliment., 1985, 74,
Anal. Chem., 1985, 68, 902. CA 103(19): 159201h. 113. CA 703(25):213508d
(54L) Tsuji, Masao; Harakawa, Mamoru; Komiyama, Y., Nippon Shokuhln (48M) Huet, Jean Claude; Pernollet, Jean Claude, J. Chromatogr., 1986,
Kogyo Gakkaishi, 1985, 32, 661. CA 704(13): 108143d. 355, 451. CA704(21): 184964n.
NITROGEN (49M) Ihekoronye, A. I., J. Food Technol., 1986, 27, 81. CA104-
(17):147276u.
(1M) Abd El-Salam, M. H.; Al-Khamy, A. F.; El-Etriby, H., Food Chem., 1986, (50M) Ihekoronye, A. I., J. Sci. Food Agric., 1985, 36, 1004. CA104-
19,213. CA 104(19): 167011g. (7):49931j.
(2M) Aiyar, K. R.; Greig, R. I. W.; Sangrouber, M. J. Food Sci., 1986, 51, (51M) Ingles, D. L.; Black, J. F.; Gallimore, D.; Tindale, R„ J. Sci. Food
856, 858. CA 105(9):77653j.
Agric., 1985, 36, 402. CA703(7):52784y.
(3M) Andrews, A. T.; Taylor, M. D.; Owen, A. J., J. Chromatogr., 1985,
(52M) Ingles, David L.; Gallimore, David, J. Chromatogr., 1985, 325, 346.
348, 177. CA104(5):33126q. CA 703(3):21332v.
(4M) Arneth, Wolfgang, Fleischwirtschaft, 1984, 64, 1086. CA 707-
(53M) James, N. A.; Ryley, Janice, J. Sci. Food Agric., 1986, 37, 151.
(25):228610p. CA 704 (17): 147291v.
(5M) Arnstadt, Klaus, Bangladesh J. Sci. Ind. Fles., 1984, 191, 210.
CA104( 13):108225g. (54M) Kaffka, K. J.; Martin, A. P„ Acfa Aliment., 1985, 74 309. CA104-
,

(6M) Banu, Laila Arjumand; Ahmed, M. Kablr; Gomes, Joseph, Bangladesh J. (15):128365f.
Sci. Ind. Fles., 1984, 19(1-4), 210-19. CA 104(23):205777w. (55M) Kane, Peter F., J. Assoc. Off. Anal. Chem., 1986, 69, 664.
CA 705(11):96135h.
(7M) Barnett, Don; Howden, Marlin E. H., Food Technol. Aust., 1984, 36,
510. CA102(3):22937n. (56M) Keller, Ronald P.; Neville, Margaret C., Clin. Chem. (Winston N.C.),
(8M) Beattie, I. G.; Games, D. E.; Startin, J.; Gilbert, J., Blomed. Mass 1986, 32(Pt. 1), 120. CA 704(11):87221k.
Spectrom., 1985, 12, 616. CA103(25):213521c. (57M) Kirst, E.; Tschop, J.; Jahn, D.; Jacobi, U., MF Milchforsch., 1985, 27,
(9M) Bender, A. E., Kiel. Milchwirtsch. Forschungsber., 1983, 35, 267. 12. CA 703(3):21300h.
CA 103(11):86568w. (58M) Kojima, Misao; Hamada, Hiroshi; Toshimitsu, Norik, Nippon Shokuhln
(10M) Bengtsson, Lena, Fette, Seifen, Anstrichm., 1985, 87, 262. CA103- Kogyo Gakkaishi, 1986, 33, 155. CA705(13):113713q.
(11):86612f. (59M) Kuninori, T.; Nishiyama, J., J. Chromatogr., 1986, 362, 255. CA105-
(11M) Bergmeyer, Hans Ulrich; Beutler, Hans Otto, Methods Enzym. Anal., (13); 113737a.
1985,8,454. Edited by Bergmeyer, H. U. CA 104(9):65052d. (60M) Lei, Mei Guey; Reeck, Gerald R„ Cereal Chem., 1986, 63, 111.
(12M) Beutler, Hans Otto, Methods Enzym. Anal., 1985, 8, 369. Edited by CA ?04(23):205801z.
Bergmeyer, H. U. CA 104 (9):65045d. (61M) Lei, Mei Guey; Reeck, Gerald R., Cereal Chem., 1986, 63, 116.
(13M) Beveridge, T.; Harrison, J. E„ Can. Inst. Food Sci. Technol. J., CA 704 (23):204040p.
1985, 18, 259. CA 103(21):177052q. (62M) Lookhart, George L.; Jones, Berne L., Cereal Chem., 1985, 62, 97.
(14M) Bican, P„ Exper/ent/a, 1985, 41, 958. CA703(13):103562j. CA 703 (7):52788c.
(15M) Bueser, W.; Erbersdobler, H. F., J. Chromatogr., 1984, 303, 234. (63M) Lookhart, G. L.; Cooper, D. B.; Jones, B. L„ Cereal Chem., 1985, 62,
CA 707(25):228620s. 19. CA 702(21):183830n.
(16M) Burns, B. Garth; Ke, Paul J., J. Assoc. Off. Anal. Chem., 1985, 68, (64M) MacDonald, John L.; Krueger, Mark W.; Keller, J., J. Assoc. Off.
444. CA 703(1):5O97s. Anal. Chem., 1985, 68, 826. CA703(19): 159200g.
(17M) Carisano, A., J. Chromatogr., 1985, 318, 132. CA102(7):60872v. (65M) Mack, D. O.; Reed, V. L.; Smith, L. D., J. Liq. Chromatogr., 1985, 8,
(18M) Carpenter, Robert N.; Brown, Rodney, J., J. Dairy Sci., 1985, 68, 591. CA 703 (5):34377p.
307. CA 702(19): 165333y.
(66M) Manji, B.; Hill, A.; Kakuda, Y.; Irvine, D. M„ J. Dairy Sci., 1985, 68,
(19M) Chang, Shyi Feu; Ayres, James W.; Sandine, W., J. Dairy Scl., 1985, 3176. CA 704 (13): 108019t.
68, 2840. CA704(5):33131n.
(67M) Matschiner, Hermann; Heberer, Henning; Guennel, G., Dev. Food Sci.,
(20M) Chong, C. S.; Kostalas, H.; Jervis, R. E., J. Radloanal. Nucl. Chem.,
1986, 120. CA 705(2): 17616q.
1986, 99, 359. CA105(6):53869h.
(68M) Matsuda, Tsukasa; Kato, Yasuko; Watanabe, Kenji; Nakamura, Ryo,
(21M) Chung, Slyin; Swaisgood, Harold E.; Catignani, G., J. Agric. Food Dev. Food Scl., 1986, 73 , 411-19. CA 705(15):132239f.
Chem., 1985, 33, 201. CA702( 13): 111548d.
(22M) Constantlnescu, B.; Ivanov, E.; Plostinaru, D.; Popa-Nemoiu, A.; Pas- (69M) Mayanna, S. M.; Jayaram, B., J. Indian Chem. Soc., 1986, 63, 329.
CA103- CA ?05(18):164220j.
covici, G., J. Radloanal. Nucl. Chem., 1985, 97, 389.
(13): 103561 h. (70M) McKillop, D. F.; Gosling, J. P.; Stevens, F. M.; Fotrell, P. F., Biochem.
(23M) Csiba, A„ Acfa Aliment., 1984, 73,189. CA707(23):209183k.
Soc. Trans., 1985, 486-7. CA703(3):21324u.
(24M) Cuq, J. L.; Cheftel, J. C., Cah. Nutr. Diet., 1985, 20, 373, 377. (71M) Menger, Anita; Baudner, S.; Guenther, H. 0., Getrelde, MeN Brot,
CA 704 (5):33079b. 1985, 39, 149. CA703(11):86591y.
(25M) Dennison, C.; Gous, R. M., S. Afr. J. Anlm. Sci., 1984, 14, 64. (72M) Moellering, Hans, Methods Enzym. Anal., 1985, 8, 350. Edited by
CA 101 (17): 149934u. Bergmeyer, H. U. CA 704(11):84573r.
(26M) Eka, O. U.; Oyeleke, O., Niger, J. Biochem., 1984, 7, 56. CA104- (73M) Molnar-Perl, I.; Plnter-Szakacs, M.; Kovago, A.; Petroczy, I.; Kralo-
(7):49914f. vanszky, U.; Matyas, J., Food Chem., 1986, 20, 21. CA104-
(27M) Ekstrand, B.; Larsson-Raznikiewicz, M., Milchwissenschaft, 1984, 39, (25):223712a.
591. CA 702(7):6O853q. (74M) Nadirov, B. T„ Vestn. SNaukiKaz., 1985, 30. CA703(15): 119268d.
(28M) Elkin, Robert G.; Griffith, Joseph, E., J. Assoc. Off. Anal. Chem., (75M) Nguyen, T. T.; Sporns, P.; Hadziyev, D., J. Chromatogr., 1986, 363,
1985, 68, 1117. CA 704(5):33160w. 361. CA 705(19):17O732u.
(29M) Elkin, Robert, G.; Griffith, Joseph E., J. Assoc. Off. Anal. Chem., (76M) Nielsen, Henrik K.; Hurrell, Richard F., J. Sci. Food Agric., 1985, 36,
1985, 68, 1028. CA 703(17): 140444t. 893. CA 704 (3): 18740s.
(30M) Elkin, Robert G„ J. Assoc. Off. Anal. Chem., 1984, 67, 1024. (77M) Oltner, Roland; Bengtsson, Staffan; Larsson, Kjel, Acta Vet. Scand.,
CA 707 (23):209211t. 1985, 26, 396. CA704(13): 108004j.
(31M) Findlay, C. J.; Parkin, K. L.; Stanley, D. W., J. Food Biochem., 1986, (78M) Osborne, B. G., Dev. Food Proteins, 1986, 4, 247. CA105-
10, 1. CA 704(17):147357w. (21):189509y.
(32M) Fischer, Beatrice; Luethy, Juerg; Schlatter, C., Z. Lebensm. Forsch., (79M) Overlay, J. C., Int. J. Appl. Radiat. Isot., 1985, 36, 185. CA103-
1984, 779, 218. CA707(23):2O9225a.
(3):21301j.
(33M) Friedman, Mendel; Noma, Amy T., J. Agric. Food Chem., 1988, 34, (80M) Paredes-Lopez, O.; Guevara, L. F., Tecnol. Aliment. (Mexico City),
497. CA 704(23):205612p.
1985,20,10. CA703(15): 121854k.
(34M) Fritschy, F.; Windemann, H.; Baumgartner, E., Z. Lebensm. Forsch.,
1985,787,379. CA 103(25):213522d. (81M) Patterson, P. L„ Lipids, 1985, 20, 503, CA703(17):140412f.
(35M) Gavlnelli, M.; Airoldi, L.f Fanelli, R., HRC CC, J. High Resolut. Chro- (82M) Peleran, J. C.; Bories, G. F., J. Chromatogr., 1985, 324, 469.
CA 703 (3):21307r.
matogr. Chromatogr. Commun., 1986, 9. CA704(25):223723e.
(36M) Gehrke, C. W.; Wall, L. L.; Absheer, J.; Kaiser, F.; Zumwalt, R., J. (83M) Perl, I. M.; Szakacs, M. P.; Kovago, A.; Petroczy, J., Food Chem.,
Assoc. Off. Anal. Chem., 1985, 68, 811. CA 103(17): 140442r. 1985 16, 163. CA 702(19): 165314t.
(37M) Grassl, Mariannae; Supp, Martin, Methods Enzym. Ana!., 1985, 7, (84M) Popineau, Y.; Pineau, Florence, Lebensm. Technol., 1985, 18, 133.
426. Edited by Bergmeyer, H. U. CA 703(21):174766q. CA 703(3):213O5p.
(38M) Green, Marie R., J. Histochem. Cytochem., 1986, 34, 147. CA104- (85M) Porras, O.; Carlsson, B.; Faellstroem, S. P.; Hanson, L., Int. Arch.
(13):108120u. Allery Appl. Immunol., 1985, 78, 30. CA 703(21):177113k.
(39M) Greiner, Steven P.; Kellen, G. J.f Carpenter, D., J. Food Sci., 1985, (86M) Raghavan, Sree K.; Ho, Chi Tang; Daun, Henryk, J. Chromatogr.,
50, 1106. CA 703(9):69891d. 1986, 351, 195. CA104(11):87220j.
(40M) Grimmer, Gernot; Naujack, Klaus Werner, J. Assoc. Off. Anal. (87M) Ray, P. K„ Indian J. Nutr. Diet., 1985, 22, 201. CA104-
Chem., 1986, 69, 537. CA705(3):23186r. (17):147283u.
(41M) Grivas, Spiros; Nyhammar, Tomas, Mutat. Res., 1985, 142, 5. (88M) Ribarova, F.; Shishkov, S„ NAHRUNG, 1986, 303, 449. CA105-
CA 702(19):165312r. (15): 132236c.

250 R • ANALYTICAL CHEMISTRY, VOL. 59, NO. 12, JUNE 15, 1987
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(89M) Ryder, John M., J. Agric. Food Chem., 1985, 33 678. CA103-
, (26N) Guilarte, Tomas R., Nutr. Rep. Int., 1985, 32, 837. CA104-
(5):36250r. (3): 18726s.
(90M) Samel, M. B. A.; Elshafie, M. A.; Hanna, M.; Csikai, J.; Juhasz, M., J. (27N) Hernandez-Mendez, J.; Alonso Mateos, A.; Almendral Parra, M. J.;
Radloanal. Ncl. Chem., 1986, 103, 81. CA 704(17): 147455b. Garcia de Maria, C., Anal. Chim. Acta, 1986, 184, 243. CA105-
(91M) Saura-Callxto, F.; Cannellas Lourdes Soler, J., An. Bromatol., 1984, (18):164225q.
1985,36,89. CA103(9):69874a. (28N) Holz, Friedhetm, Z. Lebensm. Unters. Forsch., 1984, 179, 29.
(92M) Sen, Nrlsinha P.; Seaman, Stephen W.; Kushwaha, S., Analyst (Lon- CA101(23):209169k.
don), 1986, 111, 139. CA 104(25):223685u. (29N) Hwang, Shie Ming, J. Assoc. Off. Anal. Chem., 1985, 66(4), 684-9.
(93M) Sjaunja, L. 0.; Andersson, I., Acta Agric. Scand., 1985, 35, 345. CA 103(11):86598f.
CA 104(25):223694w. (30N) Jaumann, G.; Engelhardt, H., Chromatographia, 1985, 20, 615.
(94M) Skerritt, John, H.; Diment, John A.; Wrigley, C., J. Sci. Food Agric., CA104(3):18718r.
1985, 36, 995. CA 104(7):49930h. (31N) Kneifel, W.; Sommer, R„ Ernaehrung (Vienna), 1986, 10, 459.
(95M) Skerritt, John H„ J. Scl. Food Agric., 1985, 36, 987. CA104- CA 105(19):170741w.
(7):49929q. (32N) Kneifel, Wolfgang; Sommer, Regina, Z. Lebensm. Unters. Forsch.,
(96M) Skerritt, John H.; Smith, Robyn A., J. Scl. Food Agric., 1985, 36, 1985, 181, 107. CA 103(19):159191e.
980. CA104(7):49928p. (33N) Kneifel, W„ Dtsch. Molk. Zig., 1986, 107, 212, 216. CA104-
(97M) Slump, P.; Bros, K. D., Poult. Sci., 1985, 64, 705. CA103( 1):5087p. (25):223720b.
(98M) Tomarelli, R. M.; Yuhas, R. J.; Fisher, A.; Weaber, J., J. Agric. Food (34N) Kobayashl, Tadashi; Okano, Toshio; Takeuchi, Atsuko, J. Micronutr.
Chem., 1985, 33, 316. CA102(13):111590m. Anal., 1986, 2, 1. CA105(3):23134x.
(99M) Tuffnell, Janet M.; Payne, J. W., J. Appl. Bacteriol., 1985, 58, 333. (35N) Kodaka, Kaname; Inagaki, Setsuko; Ujiie, Takashi; Ueno, Toshio;
CA102(25):219691j. Suda, Hiroyuki, Bitamln, 1985, 59, 451. CA 103(25):213475r.
(100M) Tummuru, M. K.; Sastry, C. S. P., J. Inst. Chem. (India), 1985, 57, (36N) Lambertsen, Georg, Proc. Scand. Symp. Lipids, 12th, 1983, Maruse,
167. CA 104(11):87222m. R. Ed., Llpidforum: Goeteborg, Swed, 1984. CA104( 1):4720y.
(101M) Ukeda, H.; Miyazaki, E.; Matsumoto, K.; Osajima, Y., Anal. Chem., (37N) Landen, William O., Jr., J. Assoc. Off. Anal. Chem., 1985, 68, 183.
1988, 58, 2975. CA105(21):189548k. CA102(21):183823n.
(102M) Valdes, E. V.; Summers, J. D., Poult. Scl., 1986, 65, 485. CA105- (38N) Lau, Oi Wah; Luk, Shlu Fai; Wong, Kit Sum, Analyst (London), 1986,
(1):5236c. 111,665. CA 105(17): 151620q.
(103M) Venter, B. G.; Scheepers, H. P.; Floor, J.; Snymaman, J., S. Afr. J. (39N) Lau, OI Wah; Shiu, Kwok-Keung; Chang, Shu Ting, J. Sci. Food
Dairy Technol., 1985, 17, 107. CA105( 1):5249j. Agric., 1985, 36(8), 733-739. CA103(25):213491t.
(104M) Viroben, G„ Rev. Aliment. Anim., 1985, 385, 45, 47. CA103- (40N) Lazaro, F.; Rios, A.; Luque de Castro, M.; Valcarcel, M., Analyst (Lon-
(15): 121836f. don), 1986, 111, 163. CA 104(24):213343s.
(105M) Wahlefeld, August Wilhelm; Sledel, Joachim, Methods Enzym. Ana!., (41N) Mauro, D. J.; Wetzel, D. L„ J. Chromatogr., 1984, 299(1), 281-7.
1985, 6, 488. Edited by Bergmeyer, H. U. CA 104(9):65054f. CA101(19):169218s.
(106M) Wakabayashi, K.; Takahashi, M.; Nagao, M.; Sato, S.; Kinae, N.; (42N) Mills, Robert S., J. Assoc. Off. Anal. Chem., 1985, 68, 56. CA102-
Tomita, I.; Suglmua, T., Dev. Food Scl., 1986, 363-371. CA105- (21):183836u.
(19): 170696k. (43N) Oesterdahl, B. G.; Janne, K.; Johansson, E.; Johnsson, H., Int. J.
(107M) White, W. J. P.; Lawrie, R. A., Meat Sci., 1985, 12, 117. CA102- Vttam. Nutr. Res., 1986, 56, 95. CA105( 1):5257k.
(17): 147631 q. (44N) Pachla, Lawrence A.; Reynolds, D. L.; Kissinger, P., J. Assoc. Off.
(108M) Williams, Philip C.; Mackenzie, S. L.; Starkey, P„ J. Agric. Food Anal. Chem., 1985, 68, 1. CAI02(23):202632t.
Chem., 1985, 33, 811. CA103(25):213497z. (45N) Rammell, Colin G.; Hoogenboom, Jacobus J. I., J. Liq. Chromatogr.,
(109M) Zee, J. A.; Simard, R. E.; L'Heureux, L., Lebensm. Technol., 1985, 1985,6,707. CA 103(9):67656g.
18, 245. CA 703(17): 140437t. (46N) Rychener, M.; Walter, P., Mitt. Geb. Lebensmittelunters. Hyg., 1985,
(110M) Zomborszky, Kovacs Melinda, Magy. Allatorv. Lapja, 1985, 40, 76, 112. CA 103(1):5086n.
539. CA 104 (5):33097f. (47N) Schieffer, Gary W.; Wheeler, Glenn P.; Cimino, C., J. Liq. Chroma-
togr., 1984, 7, 2659. CA 102(5):44491q.
VITAMINS (48N) Seki, Tokuichiro; Yamaguchi, Yoshihisa; Noguchi, Kohji; Yanagihara,
(1N) Aki, Hidemi; Miyamoto, Teljiro, Kaseigaku Zasshi, 1985, 36, 929. Y., J. Chromatogr., 1985, 332, 283. CA 103(19): 159217t.
CA104( 17):147281s. (49N) Sell, U., Fresenlus’ Z. Anal. Chem., 1984, 316(3-4), 287. CA101-
(2N) Ashoor, S. H.; Knox, M. J.; Olsen, J.; Deger, D. A., J. Assoc. Off. (15): 128973r.
Anal. Chem., 1985, 68, 693. CA 103(11):86599g. (50N) Sertl, David C.; Molitor, Bruce E., J. Assoc. Off. Anal. Chem., 1985,
(3N) Augustin, Jorg, J. Assoc. Off. Anal. Chem., 1984, 67, 1012. CA101- 68, 177. CA102(21):183822m.
(23):209208x. (51N) Shen, C. S. J.; Sheppard, A. J., J. Micronutr. Anal., 1986, 2, 43.
(4N) Augustine, J., Klein, B. P., Becker, D., Venugopal, P. B., Eds.; Methods CA 105(3):23166j.
of Vitamin Assay-Fourth Edition-, Wiley: New York, 1985. (52N) Speak, A. J.; Schrijver, J.; Schreurs, W. H. P., J. Food Scl., 1985,
(5N) Ayi, Benjamin K.; Yuhas, David A.; Moffett, K. S.; Joyce, D.; Deangelis, 50, 121. CAI02(13):111765x.
N. J., J. Off. Anal. Chem., 1985, 68, 1087. CA 104(7):49918k. (53N) Speek, A. J.; Temalilwa, C. R.; Schrijver, J., Food Chem., 1986, 19,
(6N) Ayi, Benjamin K.; Yuhas, David A.; Deangelis, N. J., J. Assoc. Off. 65. CA 104 (13): 108021n.
Anal. Chem., 1986, 69, 56. CA 104(11):87237v. (54N) Stancher, Bruno; Zonta, Fabio, J. Chromatogr., 1984, 312, 423.
(7N) Bltsch, R.; Salz, I.; Hoetzel, D., Dtsch. Lebensm.-Rundsch., 1986, 82, CA 102(7):60844n.
80. CA 104 (25):223700v. (55N) Trugo, L. C.; Macrae, R.; Trugo, N. M. F„ J. Micronutr. Anal., 1985,
(8N) Bognar, Antal, Z. Lebensm. Unters. Forsch., 1985, 181, 200. 1,55. CA 104 (7):49909h.
CA 103 (23): 195029s. (56N) Vanderslice, J. T.; Brownlee, S. R.; Cortissoz, M. E., J. Assoc. Off.
(9N) Bognar, Antal, Z. Lebensm. Unters. Forsch., 1986, 182, 492. Anal. Chem., 1984, 67, 999. CA 101 (25):228589p.
CA 105(11):96139n. (57N) Vanderslice, Joseph T.; Higgs, Darla J., J. Chromatogr. Sci., 1984,
(10N) Brubacher, G.; Mueller-Mulot, W.; Eds., Elsevier: London, UK, 1985; 22, 485. CA102(3):22934j.
166 pp. CA 103(19): 159399d. (58N) Velisek, Jan; Davidek, Jiri; Mnukova, J.; Pistek, T., J. Micronutr. Anal.,
(11N) Coburn, Stephen P.; Mahuren, J. Dennis, Methods Enzymol., 1986, 1986,2,73. CA105( 15):132231x.
122(Vitam. Coenzymes, Pt. G), 102. CA 105(11):93896w. (59N) Velisek, Jan; Davidek, Jiri, J. Micronutr. Anal., 1986, 2, 25. CA105-
(12N) Colugnati, Luigi, Boll. Chlm. Ig., Parte Sci., 1985, 36(S2), 65. (3):23135y.
CA103(21):177036n. (60N) Watada, Alley E.; Tran, Tony T., J. Liq. Chromatogr., 1985, 8, 1651.
(13N) Coverly, Stephen C., J. Micronutr. Anal., 1985, 1, 65. CA104- CA103( 17):140432n.
(7):49910b. (61N) Wills, R. B. H.; Wimalasiri, P.; Greenfield, H., J. Micronutr. Anal.,
(14N) Davidek, Jiri; Velisek, Jan, J. Mlcronutr. Anal., 1988, 2, 81. CA105- 1985, 1,23. CA 104 (7):49906e.
(13):113699q. (62N) Wimalasiri, P.; Wills, R. B. H„ J. Chromatogr., 1985, 318, 412.
(15N) DeLeecher, A. P.; Lambert, W. E.; DeRuyter, M. G. M., Eds., Modern CA 102(11):94420m.
Chromatographic Analysis of Vitamins-, Marcel Dekker: New York, 1985. (63N) Woollard, D. C.; Blott, A. D., J. Micronutr. Anal., 1986, 2, 97.
(16N) Echols, Richard E.; Miller, Robert H.; Thompson, L., J. Chromatogr., CA 105 (15): 132232y.
1985, 347, 89. CA103(25):213502x. (64N) Zonta, Fabio; Stancher, Bruno, J. Chromatogr., 1985, 329, 257.
(17N) Echols, Richard E.; Miller, Robert H.; Foster, William, J. Dairy Sci., CA 103(11):86587b.
1986, 69(5), 1246-1249. CA 105(7):59511g. MISCELLANEOUS
(18N) Ekanayake, Athula; Nelson, Philip E., Br. J. Nutr., 1986, 55, 235.
CA104( 17):147266r. (1P) Allen, J. C„ BNF Nutr. Bull. 1986, 11, 46. CA 104(23):205599q.
(19N) Esaka, Muneharu; Suzuki, Kanichi; Kubota, Kiyoshi, Agric. Biol. (2P) Bernardini, M.; Fedeli, E.; Baronl, D., Rlv. Ital. Sostanze Grasse, 1985,
Chem., 1985, 49, 2955. CA103(25):213510y. 62, 357. CA104(9):67550p.
(20N) Finglas, P. M.; Faulks, R. M„ Food Chem., 1984, 15, 37. CA101- (3P) Boppel, B.; Fischer, E.; Frindik, O.; Kalus, W.; Mueller, H.; Schelenz, R.,
(21): 189810g. Report, 1984. CA104(13):105094c.
(21N) Ge, Hoon; Oman, Gary N.; Ebert, Frank J., J. Assoc. Off. Anal. (4P) Charalambous, G„ Analysis of Foods and Beverages, Modern Tech-
Chem., 1986, 69, 560. CA 105(5):41347y. niques-, Academic: Orlando, FL, 1985.
(22N) Gerstenberg, Holger, Lebensmlttelchem. Oerichtl. Chem., 1985, 39, (5P) Cox, D.; Harrison, G.; Jandlk, P.; Jones, W., Food Technol. (Chicago),
1. CA 102(15):130543q. 1985, 39, 41. CA 103(17): 140400a.
(23N) Grace, Margaret L.; Bernhard, Richard A., J. Dairy Sci., 1984, 67, (6P) Davies, A. M. C.; Grant, A.; Gavrel, G.; Steeper, R., Analyst (London)
1646. CA 101 (21): 189807m. 1985,110,643. CA 104(13): 108041U.
(24N) Gregory, Jesse F., Ill, J. Assoc. Off. Anal. Chem., 1984, 67, 1015. (7P) Frankhuizen, R.; Van der Veen, N. G., Neth. Milk Dairy J. 1985, 39,
CA101(23):209131s. 191. CA104( 11):87209n.
(25N) Gregory, Jesse F., Ill; Feldstein, Debra, J. Agric. Food Chem., 1985, (8P) Giacln, Jack R.; Brzozowska, Anna, J. Plast. Film Sheeting, 1985, 1,
33(3), 359-63. CA 702(23):202672f. 292. CA 104 (23):205583e.

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(9P) Hischenhubpr, Claudia, Lebensm. Blotechnol., (Sonderh.), 1985, 4, 7. (17P) Ronalds, John A.; Miskelly, Diane, Chem. Aust., 1985, 52, 302.
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Petroleum
F. C. Trusell

Consulting Analytical Chemist, 6910 South Prince Way, Littleton, Colorado 80120

This is the 18th review in a series dating back to 1953. detecting H20, C02, CH4, and hydrocarbons during oil and
Complete bibliographic references to previous articles will be gas prospecting (IB). Colling and colleagues used multidi-
found in the 1985 review (1A). Abstracts of papers covered mensional pyrolysis gas chromatography for studying organic
in this present work appeared in Chemical Abstracts, Ana- geopolymers found in sedimentary rocks (2B). Pyrolysis GC
lytical Abstracts (London), and the American Petroleum techniques on kerogens, rocks, asphaltenes, and petroleum
Institute Refining Literature Abstracts during the period July were used by Behar (3B). Schaefer trapped and analyzed gases
1984 through June 1986. Reviews of this scope are the work from heated rock samples on an SE-54 fused silica column
of many hands. Each author is named at the beginning of using a commercial thermodesorption unit as an injection
their section. Two significant contributors, however, work in device (4B, 5B). Shang studied the geochemical characteristics
relative anonymity. C. H. Simpson, at Mobil, and D. K. and applications of aromatics in crude oil (6B, 7B). The course
Albert, at Amoco, search the abstract literature during the of biodegradation was followed by gas and liquid chroma-
biennium between reviews to provide the references reported tography detection on crude oil from the Gifhorn trough,
herein. The authors’ tasks are made much easier by their Lower Saxony region. The work was done by Teschner and
work. Wehner (8B). Seifert et al. found that surviving biomarkers
Frequently a paper appears to fit, with equal validity, into in biodegraded oils could be used for source correlation (9B).
either of two (or more) categories. The decision about where Gilmour, Swart, and Pillinger used GC and stable isotope mass
to place it is made thus: spectrometry to investigate the carbon isotope composition
If the principal thrust of the paper seems to be to char- of individual petroleum alkanes (10B). Killops and Readman
acterize the sample, the paper goes in determined aromatic hydrocarbons from oils and sediments
Section B—Crude Oil by HPLC fractionation and GC/MS (11B). GC/MS was used
Section C—Fuels, Gaseous and Liquid to analyze volatiles in fluid and gas inclusions, which provided
Section D—Lubricants, Greases, and Specialty Oils information on process history (12B). Gurko and colleagues
Section E—Asphalts, Bitumens, Tars, Pitches, and Waxes found that petroleums diffusing through geological strata
If the principal thrust of the paper seems to be to analyze fractionated (13B, 14B). Rullkoetter, Spiro, and Nissenbaum
the sample for a particular element, compound, class of com- examined the biological marker characteristics of oils from
pounds, or physical property, the paper goes in carbonate source rocks in the Dead Sea, Israel (15B). Curiale
Section F—Hydrocarbons and associates studied biomarkers in oils and rocks of the
Section G—Physical Properties Monterey Formation, California (16B), and Li and associates
Section H—Metals in Oil studied the biomarkers in source rocks of Nanyang, Damintun,
Section I—Nonmetal Elements and Compounds and Liaohe Basin in China (17B). Meyer, Christie, and Brooks
If the paper appears to emphasize the method or equipment used GC/MS with selective metastable-ion monitoring to
used, it goes in obtain biomarker information (18B, 19B). Similar work was
Section J—Analytical Methods and Apparatus also done by Philip and Gilbert (20B).
Srivastava characterized petroleum crudes of Indian origin
CRUDE OILS by NMR (2IB). Kvalheim and Telnaes also used NMR to
T. Yonko characterize Norwegian North Sea oils (22B). Zav’yalov and
Marathon Oil Company, Littleton, Colorado 80160 Razumova geochemically compared petroleums and natural
bituminoids by inrared spectroscopy (23B). Tezuka and
The petroleum industry has seen considerable changes over colleagues characterized Abu Dhabi and Japanese domestic
the past several years affecting all areas of the industry. crudes by using NMR and IR (24B). IR and X-ray diffraction
However, analytical testing in support of the petroleum in- helped in the analysis of insoluble organic matter in Domanik
dustry has not waned. Although analytical developments for rock of the Timan-Pechora region from depths <3400 m (25B).
oil shale and shale oil have decreased, new methods and Pankina and associates determined formation conditions of
techniques for petroleum and coal flourish petroleum deposits in the Timan-Pechora oil and gas provinces
The following section is divided into three major subsections (26B). Babaev and others detected characteristics of crude
(Crude Petroleum, Coal Oil, and Shade Oil). Major topics and oils from offshore fields in Azerbaijan by IR and isotopic
advances over the last few years are presented.
techniques (27B-29B). Wen found EPR to be one of the most
CRUDE PETROLEUM important tools for studying type and maturation of kerogen
(30B). Landais and Dereppe used solid-state carbon-13 NMR
Exploration. General. Mironova, Rostotskaya, and to examine Oil degradation in the Lodeve Basin, France (31B).
Naumov developed a pyrochromatographic technique for Gnoevykh, Khozyainov, and Khromov used X-ray spectros-

252 R 0003-2700/87/0359-252R$06.50/0 &copy; 1987 American Chemical Society