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Phenylpropanolamine
Molecular formula: C9H14CINO
Molecular weight: 151.2
CAS Registry No.: 154-41 -6 (phenylpropanolamine hydrochloride),
14838-15-4 (phenylpropanolamine)

SAMPLE
Matrix: blood, urine
Sample preparation: Plasma. 1 rtiL Plasma + 50 |xL 1 M NaOH + 4 mL diethyl ether,
extract, repeat extraction. Combine organic layers and evaporate them to dryness at 40
under a stream of nitrogen. Dissolve the residue in 300 |xL mobile phase A, inject a 200
IxL aliquot. Urine. 100 |JLL Urine + 1 mL 100 mM NaOH + 4 mL diethyl ether, extract,
repeat extraction. Combine organic layers and evaporate them to dryness at 40 under a
stream of nitrogen. Dissolve the residue in 300 |JLL mobile phase A, inject a 100 \xL aliquot.
Inject onto column A and elute with mobile phase A, at an appropriate time (typically,
6.5 min) the effluent from column A was eluted onto column B using mobile phase A,
when all compounds of interest had passed onto column B (typically, 8.7 min) elution of
column B was continued with mobile phase B and the effluent from column B was
monitored.
HPLCVARIABLES
Column: A 70 X 4.6 5 ^m A type YMC ODS (Yamamura); B 100 X 4.6 5 pum A type YMC
ODS (Yamamura)
Mobile phase: A MeCN: buffer 5:95 containing 5 mM sodium butanesulfonate; B MeCN:
buffer 5:95 (Buffer was 20 mM KH2PO4 adjusted to pH 3.5 with 10% orthophosphoric acid.)
Column temperature: 40
Flow rate: 1
Injection volume: 100-200
Detector: UV 205
CHROMATOGRAM
Retention time: 13
Limit of detection: 0.4 ng/mL (plasma); 8 ng/mL (urine)
KEYWORDS
plasma; column-switching; heart cut
REFERENCE
Yamashita, K.; Motohashi, M.; Yashiki, T. High-performance liquid chromatographic determination of
phenylpropanolamine in human plasma and urine, using column switching combined with ion-pair
chromatography. J.Chromatogr., 1990, 527, 103-114

SAMPLE
Matrix: bulk
Sample preparation: Prepare a 10 mg/mL solution in 500 mM sodium bicarbonate solution, extract a 10 mL aliquot twice with 15 mL portions of dichloromethane. Combine the
extracts and add 10 |xL phenylisothiocyanate, evaporate to dryness under a stream of air,
reconstitute with 10 mL MeOH, inject a 10 |xL aliquot.
HPLCVARIABLES
Guard column: 70 X 2.1 CO:PELL ODS
Column: 300 X 3.9 ixBondapak C18

Mobile phase: MeOH: water: acetic acid 45:54:1

Flow rate: 2
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 10
OTHER SUBSTANCES

Simultaneous: ephedrine, lidocaine, pseudoephedrine

KEYWORDS

derivatization
REFERENCE
Noggle, F.T., Jr.; Clark, CR. Liquid chromatographic analysis of samples containing cocaine, local anesthetics, and other amines. J.Assoc.Off.Anal.Chem., 1983, 66, 151-157

SAMPLE

Matrix: formulations
Sample preparation: Grind ten tablets to a fine powder, add 15 mL MeCN: buffer 50:50,
sonicate for 15 min, make up to 20 mL with MeCN: buffer 50:50, filter, dilute tenfold,
inject a 10 |xL aliquot. (Buffer was 50 mM sodium perehlorate adjusted to pH 3.0 with
70% perchloric acid.)
HPLCVARIABLES

Column: 250 X 4 10 |xm Lichrosorb RP-18

Mobile phase: Gradient. MeCN: buffer from 15:85 to 95:5 over 20 min, re-equilibrate at
initial conditions for 10 min. (Buffer was 50 mM sodium perehlorate adjusted to pH 3.0
with 70% perchloric acid.)
Column temperature: 25
Flow rate: 2
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 2.5
OTHER SUBSTANCES

Simultaneous: buzepide (UV 220), clocinizine

KEYWORDS

tablets
REFERENCE
Cavazzutti, G.; Gagliardi, L.; De Orsi, D.; Tonelli, D. Simultaneous determination of buzepide, phenylpropanolamine, and clocinizine on pharmaceutical preparations by ion-pair reversed-phase HPLC.
J.Liq.Chromatogr., 1995, 18, 227-234

SAMPLE

Matrix: formulations
Sample preparation: Finely powder half a tablet, add 9 mL mobile phase, sonicate for 20
min, make up to 10 mL with mobile phase, filter (Whatman type 40 and 0.2 |xm Millipore),
inject an aliquot of the filtrate.

HPLCVARIABLES
Column: 250 X 4 5 jxm LiChrospher 100 CN
Mobile phase: MeCN:THF:buffer 7:6:87 (Buffer was 0.8% acetic acid containing 5 mM
sodium hexanesulfonate, 10 mM di-n-butylamine, and 0.12% phosphoric acid, pH 3.3.)
Flow rate: 1
Injection volume: 20
Detector: UV 260
CHROMATOGRAM
Retention time: 3.5
Limit of detection: 15.1 |xg/mL
OTHER SUBSTANCES
Simultaneous: acetaminophen (UV 310), caffeine (UV 298), chlorpheniramine (UV 265),
guaifenesin (glycerylguaicolate) (UV 284)
KEYWORDS
tablets
REFERENCE
Indrayanto, G.; Sunarto, A.; Adriani, Y. Simultaneous assay of phenylpropanolamine hydrochloride,
caffeine, paracetamol, glycerylguaiacolate and chlorpheniramine in Silabat tablet using HPLC with
diode array detection. J.Pharm.Biomed.AnaL, 1995, 13, 1555-1559

SAMPLE
Matrix: formulations
Sample preparation: Dilute 10 mL to 50 mL with water, inject an aliquot.
HPLCVARIABLES
Column: 250 X 4.6 Whatman 10 jxm PXS SCX
Mobile phase: MeOH: 100 mM (NH4)H2PO4, apparent pH 6.2
Column temperature: 40
Flow rate: 2
Injection volume: 20
Detector: UV 263
CHROMATOGRAM
Retention time: 3.12
OTHER SUBSTANCES
Simultaneous: dextromethorphan
KEYWORDS
liquid formulations; stability-indicating
REFERENCE
Wilson, T.D.; Jump, W.G.; Neumann, W.C.; San Martin, T. Validation of improved methods for highperformance liquid ehromatographic determination of phenylpropanolamine, dextromethorphan,
guaifenesin and sodium benzoate in a cough-cold formulation. J.Chromatogr., 1993, 641, 241-248

SAMPLE
Matrix: formulations
Sample preparation: Tablets. One tablet + 50 mL MeOH, sonicate, make up to 100 mL
with MeOH, centrifuge for 15 min. Remove 1 mL supernatant, make up to 10 mL with
mobile phase, inject a 50 |xL aliquot. Drops. Dilute drops with the mobile phase so that

the concentration of phenylpropanolamine hydrochloride is 50 jjig/mL, inject a 50 JULL

aliquot.
HPLCVARIABLES
Column: 100 X 4.6 Cyclobond I (Advanced Separation Technologies)
Mobile phase: MeOH: 50 mM NaH2PO4 adjusted to pH 7.0 with 0.1 M NaOH 30:70
Column temperature: 35
Flow rate: 1.5
Injection volume: 50
Detector: UV 254
CHROMATOGRAM
Retention time: 1.8
OTHER SUBSTANCES
Simultaneous: pheniramine, pyrilamine (mepyramine)
KEYWORDS
tablets; drops
REFERENCE
el-Gizawy, S.M.; Ahmed, A. High-performance liquid chromatographic determination of mepyramine
maleate, pheniramine maleate and phenylpropanolamine hydrochloride in tablets and drops. Analyst, 1987, 112, 867-869

SAMPLE
Matrix: formulations
Sample preparation: Injections. Dilute 1.5 mL of a 20 mg/mL injection to 100 mL with
water, remove a 10 mL aliquot and add it to 8 mL 2% phenylpropanolamine hydrochloride,
make up to 100 mL with water, inject a 20 |xL aliquot. Tablets. Grind tablets to a fine
powder, weigh out amount equivalent to about 10 mg hydralazine, mix thoroughly with
2 mL 500 mM HCl, make up to 100 mL with water, shake for 2-3 min, filter, discard first
15 mL. 15 mL Filtrate + 4 mL 2% phenylpropanolamine hydrochloride, make up to 50
mL with water, inject a 20 |xL aliquot.
HPLCVARIABLES
Column: 300 X 3.9 ixBondapak C18
Mobile phase: MeOH: 15 mM KH2PO4: glacial acetic acid 2:97.9:0.1
Flow rate: 3
Injection volume: 20
Detector: UV 256
CHROMATOGRAM
Retention time: 3
Internal standard: phenylpropanolamine
OTHER SUBSTANCES
Simultaneous: hydralazine, hydrochlorothiazide
KEYWORDS
injections; tablets; phenylpropanolamine is IS
REFERENCE
Das Gupta, V. Quantitation of hydralazine hydrochloride in pharmaceutical dosage forms using highperformance liquid chromatography. J.Liq.Chromatogr., 1985, 8, 2497-2509

SAMPLE

Matrix: formulations
Sample preparation: Grind tablets to a fine powder and weigh out powder equivalent to
about 80 mg metronidazole, add 90 mL water, heat to 60 with stirring, cool to room
temperature, make up to 100 mL with water, filter, reject first 20 mL of filtrate. Mix 12.5
mL filtrate with 7 mL 10 mg/mL phenylpropanolamine hydrochloride in water, make up
to 50 mL with water, inject a 20 |xL aliquot.
HPLCVARIABLES

Column: 300 X 4 10 |xm jxBondapak phenyl

Mobile phase: 20 mM KH2PO4, pH 4.2
Flow rate: 2.5
Injection volume: 20
Detector: UV 254
CHROMATOGRAM

Retention time: 3.5

Internal standard: phenylpropanolamine hydrochloride
OTHER SUBSTANCES

Simultaneous: metronidazole
Noninterfering: excipients, mannitol
KEYWORDS

tablets; stability-indicating; phenylpropanolamine is IS

REFERENCE
Das Gupta, V. Quantitation of metronidazole in pharmaceutical dosage forms using high-performance
liquid chromatography. J.Pharm.ScL, 1984, 73, 1331-1333

SAMPLE

Matrix: formulations
Sample preparation: Leach 200 or 300 mg ground capsule or tablet with water or mobile
phase and dilute to 50 mL, sonicate for 5 min, centrifuge at 2500 rpm for 5 min, inject
an aliquot. Dilute 4-25 mL of liquid formulations to 250 mL with water, inject an aliquot.
HPLCVARIABLES

Column: Partisil-10 C8
Mobile phase: MeOH:MeCN:water:PIC-B5 50:170:755:25 (PIC-B5 (Waters) is 200 mM
sodium pentanesulfonate in glacial acetic acid.)
Flow rate: 2
Injection volume: 20
Detector: UV 254
CHROMATOGRAM

Retention time: 4.5

OTHER SUBSTANCES

Simultaneous: impurities, degradation products, benzoic acid, guaifenesin, phenylephrine

KEYWORDS

tablets; capsules; liquid formulations; stability-indicating

REFERENCE
Schieffer, G.W.; Smith, W.O.; Lubey, G.S.; Newby, D.G. Determination of the structure of a synthetic
impurity in guaifenesin: modification of a high-performance liquid chromatographic method for
phenylephrine hydrochloride, phenylpropanolamine hydrochloride, guaifenesin, and sodium benzoate
in dosage forms. J.Pharm.ScL, 1984, 73, 1856-1858

SAMPLE

Matrix: formulations
Sample preparation: Capsules and Tablets. Leach 1 g of ground capsule or tablet with
250 mL 0.4 mg/mL 2,5-dihydroxybenzoic acid in water, sonicate for 10 min, centrifuge at
2500 rpm for 5 min, inject an aliquot. Liquid formulations. Dilute 4-25 mL of the formulation to 250 mL with 0.4 mg/mL 2,5-dihydroxybenzoic acid in water, inject an aliquot.
HPLCVARIABLES

Column: 250 X 4.6 Partisil 10 C8

Mobile phase: MeOH:water:PIC-B5 300:675:25 (PIC-B5 (Waters) is 200 mM sodium pentanesulfonate in glacial acetic acid.)
Flow rate: 2
Injection volume: 20
Detector: UV 254
CHROIUIATOGRAM

Retention time: 6
Internal standard: 2,5-dihydroxybenzoic acid (4.5)
OTHER SUBSTANCES

Simultaneous: impurities, degradation products, guaifenesin, phenylephrine

KEY WORDS

capsules; tablets; liquid formulations; stability-indicating

REFERENCE
Schieffer, G.W.; Hughes, D.E. Simultaneous stability-indicating determination of phenylephrine hydrochloride, phenylpropanolamine hydrochloride, and guaifenesin in dosage forms by reversed-phase
paired-ion high-performance liquid chromatography. J.Pharm.Sci., 1983, 72, 55-59

SAMPLE

Matrix: formulations
Sample preparation: Crush 10 tablets, add 250 mL 50 mM HCl in EtOH: water 50:50,
heat for 15 min on a steam bath, shake mechanically for 2 h, filter (glass fiber GF/A,
Whatman), inject a 30 |xL aliquot of the filtrate.
HPLCVARIABLES

Column: 250 x 4.6 10 |xm Partisil- 10-ODS

Mobile phase: MeCN: buffer 50:50 (Buffer was 2.85 mM ethylenediamine sulfate adjusted
to pH 7.44 0.02 with 1 M ammonium hydroxide.)
Flow rate: 3.8
Injection volume: 30
Detector: UV 216.5
CHROMATOGRAM
Retention time: 4
OTHER SUBSTANCES

Simultaneous: aposcopolamine, methscopolamine, pheniramine, pyrilamine, tropic acid

KEYWORDS

tablets
REFERENCE
Heidemann, D.R. High-pressure liquid chromatographic determination of methscopolamine nitrate,
phenylpropanolamine hydrochloride, pyrilamine maleate, and pheniramine maleate in tablets.
J.Pharm.ScL, 1981, 70, 820-822

SAMPLE

Matrix: solutions
HPLC VARIABLES

Guard column: 30 X 2.1 Spheri-5 RP-8

Column: 220 X 2.1 Spheri-5 RP-8
Mobile phase: Gradient. A was 0.08% diethylamine and 0.09% phosphoric acid in water,
pH 2.3. B was MeCN: water 90:10 containing 0.08% diethylamine and 0.09% phosphoric
acid. A:B 95:5 for 2 min, to 0:100 over 15 min (?), maintain at 0:100 for 5 min.
Column temperature: 50
Flow rate: 0.5
Detector: UV 200
CHROMATOGRAM

Retention time: 4.5

OTHER SUBSTANCES

Simultaneous: amphetamine, diethylpropion, ephedrine, fenfluramine, methamphetamine, phentermine

Also analyzed: amitriptyline, chlordiazepoxide, chlorpromazine, desalkylflurazepam, desipramine, desmethyldoxepin, diazepam, doxepin, flurazepam, imipramine, mesoridazine,
norchlordiazepoxide, nordiazepam, nortriptyline, oxazepam, prazepam, promazine, thioridazine, thiothixene, trifluoperazine
REFERENCE
Rainin Catalog Ci-94, 1994-5, Rainin Instrument Co., Woburn MA, p. 7.24

SAMPLE

Matrix: solutions
HPLCVARIABLES

Column: 250 X 4.6 Zorbax RX

Mobile phase: Gradient. A was 10 mL concentrated orthophosphoric acid and 7 mL triethylamine in 1 L water. B was 10 mL concentrated orthophosphoric acid and 7 mL triethylamine in 200 mL water, make up to 1 L with MeCN. A:B from 100:0 to 0:100 over
30 min, maintain at 0:100 for 5 min.
Column temperature: 30
Flow rate: 2
Detector: UV 210
OTHER SUBSTANCES

Also analyzed: acepromazine, acetaminophen, acetophenazine, albuterol, aminophylline,

amitriptyline, amobarbital, amoxapine, amphetamine, amylocaine, antipyrine, aprobarbital, aspirin, atenolol, atropine, avermectin, barbital, benzocaine, benzoic acid, benzotropine, benzphetamine, berberine, bibucaine, bromazepan, brompheniramine, buprenorphine, buspirone, butabarbital, butacaine, butethal, caffeine, carbamazepine, carbromal,
chloramphenicol, chlordiazepoxide, chloroquine, chlorothiazide, chloroxylenol, chlorphe-

nesin, chlorpheniramine, chlorpromazine, chlorpropamide, chlortetracycline, cimetidine,

cinchonidine, cinchonine, clenbuterol, clonazepam, clonixin, clorazepate, cocaine, codeine,
colchicine, cortisone, coumarin, cyclazocine, cyclobenzaprine, cyclothiazide, cyheptamide,
cymarin, danazol, danthron, dapsone, debrisoquine, desipramine, dexamethasone, dextromethorphan, dextropropoxyphene, diamorphine, diazepam, diclofenac, diethylpropion,
diethylstilbestrol, diflunisal, digitoxin, digoxin, diltiazem, diphenhydramine, diphenoxylate, diprenorphine, dipyrone, disulfiram, dopamine, doxapram, doxepin, dronabinol,
ephedrine, epinephrine, epinine, estradiol, estriol, estrone, ethacrynic acid, ethosuximide,
etonitazene, etorphine, eugenol, famotidine, fenbendazole, fencamfamine, fenoprofen, fenproporex, fentanyl, flubendazole, flufenamic acid, flunitrazepam, 5-fluorouracil, fluoxymesterone, fluphenazine, furosemide, gentisic acid, gitoxigenin, glipizide, glunixin,
glutethimide, glybenclamide, guaiacol, halazepam, haloperidol, hydrochlorothiazide,
hydrocodone, hydrocortisone, hydromorphone, hydroxyquinoline, ibogaine, ibuprofen, iminostilbene, imipramine, indomethacin, isocarbostyril, isocarboxazid, isoniazid, isoproterenol, isoxsuprine, ivermectin, ketamine, ketoprofen, kynurenic acid, levorphanol, Iidocaine, lorazepam, lormetazepam, loxapine, mazindol, mebendazole, meclizine,
meclofenamic acid, medazepam, mefenamic acid, megestrol, mepacrine, meperidine, mephentermine, mephenytoin, mephesin, mephobarbital, mepivacaine, mescaline, mesoridazine, methadone, methamphetamine, methapyrilene, methaqualone, methazolamide,
methocarbamol, methoxamine, methsuximide, methyl salicylate, methyldopa, methyldopamine, methylphenidate, methylprednisolone, methyltestosterone, methyprylon, metoprolol, mibolerone, morphine, nadolol, nalorphine, naloxone, naltrexone, naphazoline,
naproxen, nefopam, niacinamide, nicotine, nicotinic acid, nifedipine, niflumic acid, nitrazepam, norepinephrine, nortriptyline, noscapine, nylidrin, oxazepam, oxycodone, oxymorphone, oxyphenbutazone, oxytetracycline, papaverine, pargyline, pemoline, pentazocine,
pentobarbital, persantine, phenacetin, phenazocine, phenazopyridine, phencyclidine,
phendimetrazine, phenelzine, pheniramine, phenobarbital, phenothiazine, phensuximide,
phentermine, phenylbutazone, phenylephrine, piperocaine, prazepam, prednisolone,
primidone, probenecid, progesterone, propiomazine, propranolol, propylparaben, pseudoephedrine, puromycin, pyrilamine, pyrithyldione, quazepam, quinaldic acid, quinidine, quinine, ranitidine, recinnamine, reserpine, resorcinol, saccharin, albuterol, salicylamide,
salicylic acid, scopolamine, scopoletin, secobarbital, strychnine, sulfacetamide, sufadiazine, sulfadimethoxine, sulfaethidole, sulfamerazine, sulfamethazine, sulfamethoxizole,
sulfanilamide, sulfapyridine, sulfasoxizole, sulindac, tamoxifen, temazepam, testosterone,
tetracaine, tetracycline, tetramisole, thebaine, theobromine, theophylline, thiabendazole,
thiamine, thiamylal, thiobarbituric acid, thioridazine, thiosalicylic acid, thiothixene, thymol, tolazamide, tolazoline, tobutamide, tolmetin, tranylcypromine, triamcinolone, tribenzylamine, trichloromethiazide, trifluoperazine, trihexyphenidyl, trimethoprim, tripelennamine, triprolidine, tropacocaine, tyramine, verapamil, vincamine, warfarin, yohimbine,
zoxazolamine
REFERENCE
Hill, D.W.; Kind, A. J. Reversed-phase solvent-gradient HPLC retention indexes of drugs. J.Anal. ToxicoL,
1994, 18, 233-242

SAMPLE
Matrix: solutions
HPLCVARIABLES
Column: 150 X 4.6 Supelcosil LC-ABZ
Mobile phase: MeCN: 25 mM pH 6.9 potassium phosphate buffer 35:65
Flow rate: 1.5
Injection volume: 25
Detector: UV 254
CHROMATOGRAM
Retention time: 1.312

OTHER SUBSTANCES

Also analyzed: 6-acetylmorphine, amiloride, amphetamine, benzocaine, benzoylecgonine,

caffeine, cocaine, codeine, doxylamine, fluoxetine, glutethimide, hexobarbital, hypoxanthine, levorphanol, LSD, meperidine, mephobarbital, methadone, methylphenidate, methyprylon, N-norcodeine, oxazepam, oxycodone, prilocaine, procaine, terfenadine
REFERENCE
Ascah, T.L. Improved separations of alkaloid drugs and other substances of abuse using Supelcosil LCABZ column. Supelco Reporter, 1993, 12(3), 18-21

SAMPLE

Matrix: solutions
Sample preparation: Prepare a 0.5 mg/mL solution in water, inject a 5 JJLL aliquot.
HPLCVARIABLES

Column: 250 X 4.6 Zorbax RX

Mobile phase: Gradient. A was 150 mM phosphoric acid and 50 mM triethylamine. B was
MeCN:water 80:20 containing 150 mM phosphoric acid and 50 mM triethylamine. A:B
100:0 for 2.2 min then to 0:100 over 30 min.
Column temperature: 30
Flow rate: 2
Injection volume: 5
Detector: UV 210
CHROMATOGRAM
Retention time: 4.9
OTHER SUBSTANCES

Simultaneous: acetaminophen, aprobarbital, butabarbital, chlordiazepoxide, chloroxylenol,

chlorpromazine, clenbuterol, cortisone, danazol, diflunisal, doxapram, estrone, fluoxymesterone, mefenamic acid, methyltestosterone, nicotine, oxazepam, phentermine, progesterone, sulfamethazine, sulfanilamide, testosterone, testosterone propionate, tranylcypromine, tripelennamine
REFERENCE
Hill, D.W.; Kind, A.J. The effects of type B silica and triethylamine on the retention of drugs in silica
based reverse phase high performance chromatography. J.Liq.Chromatogr., 1993, 16, 39413964

SAMPLE

Matrix: solutions
Sample preparation: Prepare a 1.5 mg/mL solution, inject a 10 |xL aliquot.
HPLCVARIABLES

Guard column: Supelguard LC-8-DB (Supelco)

Column: 50 x 4.6 Supelcosil LC-8-DB
Mobile phase: MeCN: buffer 10:90 containing 0.02% triethylamine (Buffer was KH2PO4
adjusted to pH 2.0 with phosphoric acid.)
Column temperature: 35
Flow rate: 2
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 1

OTHER SUBSTANCES
Simultaneous: chlorpheniramine, methscopolamine, pseudoephedrine, triprolidine
REFERENCE
Supelco Catalog, Supelco, Inc., Bellefonte MA, 1992, p. 179

SAMPLE
Matrix: solutions
Sample preparation: Dissolve in MeOH: water 1:1 at a concentration of 50 jxg/mL, inject
a 10 jxL aliquot.
HPLC VARIABLES
Column: 300 X 3.9 10 |xm jxBondapak C18
Mobile phase: MeOH: acetic acid: triethylamine: water 5:1.5:0.5:93
Flow rate: 1.5
Injection volume: 10
Detector: UV 254
CHROMATOGRAM
Retention time: 7
OTHER SUBSTANCES
Simultaneous: ephedrine, pseudoephedrine
REFERENCE
Roos, R.W.; Lau-Cam, CA. General reversed-phase high-performance liquid chromatographic method
for the separation of drugs using triethylamine as a competing base. J.Chromatogr., 1986, 370,
403-418

SAMPLE
Matrix: solutions
Sample preparation: Dissolve in MeOH at a concentration of 1 mg/mL, inject a 20 |xL
aliquot.
HPLCVARIABLES
Column: 250 X 5 Spherisorb S5W
Mobile phase: MeOH:buffer 90:10 (Buffer was 94 mL 35% ammonia and 21.5 mL 70%
nitric acid in 884 mL water, adjust the pH to 10.1 with ammonia.)
Flow rate: 2
Injection volume: 20
Detector: UV 254
CHROMATOGRAM
Retention time: 2.25
OTHER SUBSTANCES
Simultaneous: amphetamine, benzphetamine, benzylmorphine, bromo-STP, buprenorphine, caffeine, codeine, codeine-N-oxide, dextromoramide, dextropropoxyphene, diethylpropion, dihydrocodeine, dihydromorphine, dimethylamphetamine, ephedrine, ethoheptazine, ethylmorphine, etorphine, fenethyline, fenfluramine, fentanyl, hydrocodone,
4-hydroxyamphetamine, hydroxypethidine, levallorphan, levorphanol, mazindol, mephentermine, mescaline, methadone, methamphetamine, methylenedioxyamphetamine, methylephedrine, methylphenidate, morphine, morphine-3-glucuronide, morphine-N-oxide,
nalorphine, naloxone, norcodeine, norlevorphanol, normetanephrine, normethadone, normorphine, norpethidine, norpipanone, noscapine, papaverine, pemoline, phenazocine,
phendimetrazine, phenelzine, 2-phenethylamine, phenoperidine, phenylephrine, pholcodeine, piritramide, prolintane, pseudoephedrine, STP, thebaine, tranylcypromine, trimethoxyamphetamine, tyramine

Noninterfering: dopamine, levodopa, methyldopa, methyldopate, norepinephrine

Interfering: acetylcodeine, chlorphentermine, diamorphine, dipipanone, epinephrine, fencamfamin, meperidine, monoacetylmorphine, norpseudoephedrine, oxycodone, pentazocine, phentermine, pipradol, thebacon
REFERENCE
Law, B.; Gill, R.; Moffat, A.C. High-performance liquid chromatography retention data for 84 basic drugs
of forensic interest on a silica column using an aqueous methanol eluent. J.Chromatogr., 1984, 301,
165-172

SAMPLE
Matrix: urine
Sample preparation: 1 mL Urine + 0.5 mL 1% trichloroacetic acid, centrifuge at 5200 g
for 10 min, filter (0.2 |xm), inject 20 |xL aliquot.
HPLCVARIABLES
Column: 250 X 4 Lichrospher 5 |xm 60 RP-select B
Mobile phase: Gradient. MeCN: 50 mM pH 3.2 potassium phosphate buffer from 10:90 to
50:50 over 15 min.
Flow rate: 1.5
Injection volume: 20
Detector: UV 190-370
CHROMATOGRAM
Retention time: 4
OTHER SUBSTANCES
Extracted: benzoylecgonine, cocaine, diphenhydramine, ephedrine, lidocaine, morphine,
nordiazepam, norpropoxyphene, nortriptyline
Also analyzed: amitriptyline, amphetamine, codeine (different gradient), meperidine
REFERENCE
Li, S.; Gemperline, RJ.; Briley, K.; Kazmierczak, S. Identification and quantitation of drugs of abuse in
urine using the generalized rank annihilation method of curve resolution. J.Chromatogr.B, 1994,
655, 213-223

SAMPLE
Matrix: urine
Sample preparation: 5 mL Urine + 25 |xL mobile phase + 100 |xL 10 M NaOH + 2 mL
diethyl ether + 3 g sodium sulfate, shake for 20 min, centrifuge at 1200 g for 5 min.
Remove the organic layer and evaporate it to dryness under a stream of nitrogen, reconstitute the residue in 100 |xL mobile phase, inject a 20 |xL aliquot.
HPLCVARIABLES
Column: 125 X 4 5 |jim LiChrospher 60 RP Select B
Mobile phase: 200 mM pH 5.5 phosphate buffer containing 150 mM triethylamine (Wash
column with MeOH for 15 min and with water for 15 min at the end of each day.)
Column temperature: 40
Flow rate: 1.3
Injection volume: 20
Detector: UV 215
CHROMATOGRAM
Retention time: 11.5
Internal standard: phenylpropanolamine
Limit of detection: 500 ng/mL

OTHER SUBSTANCES
Extracted: ephedrine, ethylephedrine, N-methylephedrine, norephedrine, norpseudoephedrine, pseudoephedrine
Noninterfering: amfepramone, amphetamine, caffeine, chlorphentermine, cocaine, codeine, cropropamide, crotethamide, dimethylamphetamine, etamivan, fencamfamine,
heptaminol, leptazol, lidocaine, meperidine, methoxamine, methylamphetamine, methylphenidate, nicotine, niketamine, phendimetrazine, phenmetrazine, pipradol, procaine,
prolintane, strychnine
KEYWORDS
phenylpropanolamine is IS
REFERENCE
Imaz, C; Carreras, D.; Navajas, R.; Rodriguez, C; Rodriguez, A.F.; Maynar, J.; Cortes, R. Determination
of ephedrines in urine by high-performance liquid chromatography. J.Chromatogr., 1993, 631, 2 0 1 205

SAMPLE
Matrix: urine
Sample preparation: 500 |xL Urine + chlorpheniramine + 100 |xL buffer, centrifuge at
11000 g for 30 s, inject a 500 |xL aliquot onto column A with mobile phase A, after 0.6
min backflush column A with mobile phase A to waste for 1.6 min, elute column A with
250 |xL mobile phase B, with 200 |JLL mobile phase C, and with 1.15 mL mobile phase D.
Elute column A to waste until drugs start to emerge then elute onto column B. Elute
column B to waste until drugs start to emerge, then elute onto column C. When all the
drugs have emerged from column B, remove it from the circuit. Elute column C with
mobile phase D, monitor the effluent from column C. Flush column A with 7 mL mobile
phase E, mobile phase D, and mobile phase A. Flush column B with 5 mL mobile phase
E then with mobile phase D. (Buffer was 6 M ammonium acetate adjusted to pH 8.0 with
2 M KOH.)
HPLCVARIABLES
Column: A 10 X 2.1 12-20 |xm PRP-I spherical poly(styrene-divinylbenzene) (Hamilton); B
10 X 3.2 11 jim Aminex A-28 (Bio-Rad); C 25 X 3.2 5 |xm C8 (Phenomenex) + 150 X 4.6
5 |xm silica (Macherey-Nagel)
Mobile phase: A 0.1% pH 8.0 potassium borate buffer; B 6 mM KH2PO4 containing 5 mM
tetramethylammonium hydroxide, and 2 mM dimethyloctylamine, pH adjusted to 6.50
with phosphoric acid; C MeCN: buffer 40:60 (Buffer was 6 mM KH2PO4 containing 5 mM
tetramethylammonium hydroxide, and 2 mM dimethyloctylamine, pH adjusted to 6.50
with phosphoric acid.); D MeCN: buffer 33:67 (Buffer was 6 mM KH2PO4 containing 5
mM tetramethylammonium hydroxide, and 2 mM dimethyloctylamine, pH adjusted to
6.50 with phosphoric acid.); E MeCN:buffer 70:30 (Buffer was 6 mM KH2PO4 containing
5 mM tetramethylammonium hydroxide, and 2 mM dimethyloctylamine, pH adjusted to
6.50 with phosphoric acid.)
Column temperature: 40 (B, C)
Flow rate: A 5; B-E 1
Injection volume: 500
Detector: UV 210; UV 235
CHROMATOGRAM
Retention time: k' 2.2
Internal standard: chlorpheniramine (k' 5.9)
Limit of detection: 300 ng/mL

OTHER SUBSTANCES
Extracted: amitriptyline, benzoylecgonine, caffeine, codeine, cotinine, desipramine, diazepam, diphenhydramine, ephedrine, flurazepam, hydrocodone, hydromorphone, imipramine, lidocaine, methadone, methamphetamine, morphine, nordiazepam, nortriptyline,
oxazepam, pentazocine, phenmetrazine, phenobarbital, secobarbital
Interfering: amphetamine, phentermine
KEYWORDS
column-switching
REFERENCE
Binder, S.R.; Regalia, M.; Biaggi-McEachern, M.; Mazhar, M. Automated liquid chromatographic analysis of drugs in urine by on-line sample cleanup and isocratic multi-column separation.
J.Chromatogr., 1989, 473, 325-341

ANNOTATED BIBLIOGRAPHY
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Stockley, CS.; Wing, L.M.; Miners, J.O. Stereospecific high-performance liquid chromatographic assay
for the enantiomers of phenylpropanolamine in human plasma. Ther.Drug Monit., 1991, 13, 332338
Shi, R.J.Y.; Gee, W.L.; Williams, R.L.; Benet, L.Z.; Lin, E.T. Ion-pair liquid chromatographic analysis of
phenylpropanolamine in plasma and urine by post-column derivatization with o-phthalaldehyde.
J.Liq.Chromatogr., 1985, 8, 1489-1500 [post-column reaction; fluorescence detection; LOD 2 ng/mL;
a-methylbenzylamine (IS)]
Costanzo, S. J. Selection of mixed ion-pair modifiers for high-performance liquid chromatographic mobile
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Das Gupta, V.; Heble, A.R. Quantitation of acetaminophen, chlorpheniramine maleate, dextromethorphan hydrobromide, and phenylpropanolamine hydrochloride in combination using high-performance
liquid chromatography. J.P/iarra.ScL, 1984, 73, 1553-1556
Dye, D.; East, T.; Bayne, WF. High-performance liquid chromatographic method for post-column, in-line
derivatization with o-phthalaldehyde and fluorometric detection of phenylpropanolamine in human
urine. J.Chromatogr., 1984, 284, 457-461
Dowse, R.; Haigh, J.M.; Kanfer, I. Determination of phenylpropanolamine in serum and urine by highperformance liquid chromatography. J.Pharm.ScL, 1983, 72, 1018-1020
Tan, H.S.; Salvador, G.C Assay of mixtures of phenylpropanolamine hydrochloride and caffeine in appetite suppressant formulations by high-performance liquid chromatography. J.Chromatogr., 1983,
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Lurie, LS. Improved isocratic mobile phases for the reverse phase ion-pair chromatographic analysis of
drugs of forensic interest. J.Liq.Chromatogr., 1981, 4, 399-408 [also amobarbital, amphetamine,
butabarbital, caffeine, diamorphine, ephedrine, methamphetamine, pentobarbital, phenobarbital,
phentermine, secobarbital]
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acetaminophen, acetylcodeine, acetylmorphine, aminopyrene, aminopyrine, amobarbital, amphetamine, antipyrine, benzocaine, butabarbital, caffeine, cocaine, codeine, diamorphine, diazepam, diethylpropion, DMT, ephedrine, glutethimide, Lampa, lidocaine, LSD, MDA, mecloqualone, mescaline,
methamphetamine, methapyrilene, methaqualone, methpyrilene, methylphenidate, morphine, narcotine, papaverine, PCP, pentobarbital, phencyclidine, phendimetrazine, phenmetrazine, phenobarbital, phentermine, procaine, quinidine, quinine, secobarbital, strychnine, TCP, tetracaine, thebaine,
theophylline]
Mason, WD.; Amick, E.N. High-pressure liquid chromatographic analysis of phenylpropanolamine in
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Achari, R.G.; Jacob, J.T. A study of the retention behavior of some basic drug substances by ion-pair
HPLC. J.Liq.Chromatogr., 1980, 3, 81-92 [also N-acetylprocainamide, antazoline, atropine, caffeine,
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Muhammad, N.; Bodnar, J.A. Quantitative determination of guaifenesin, phenylpropanolamine hydrochloride, sodium benzoate & codeine phosphate in cough syrups by high-pressure liquid chromatography. J.Liq.Chromatogr., 1980, 3, 113-122