P-Anisidine Value

Anisidine value is the number of secondary products of lipid compounds oxidation

which are formed due to breakdown of peroxides and hydroperxides.

From: Food Chemistry, 2018

Related terms:

Wheat Flour, Doughnut, Batter, Sweet, Whipped Cream, Raisin

View all Topics

Measuring lipid oxidation

J.W. Irwin, N. Hedges, in Understanding and Measuring the Shelf-Life of Food, 2004

13.2.3 Anisidine value

Anisidine value (AV) is a measure of the aldehyde levels in an oil or fat, in particular
those that are unsaturated (and principally the 2–alkenals). To determine AV, a
solution of the oil or fat in iso-octane is reacted with p-anisidine in glacial acetic acid
to form yellowish reaction products. The AV is then determined from the absorbance
measured at 350 nm, both before and after reaction. By convention, AV is defined
as 100 times the increase in absorbance measured at 350 nm in a 1 cm cell of a
solution resulting from the reaction of 1 g oil or fat in 100 ml solvent with anisidine
reagent. The method is not suitable for highly coloured oils, particularly those that
have a high absorbance at this wavelength.

As a measure of secondary oxidation products, AV is used instead of, or together with,

PV to assess thermally stressed oils. It may be used, for instance, in studies carried
out to maximise the life of frying oils.14 AV may also be used to support sensory data,
as in an investigation of the storage of tortilla chips fried in various oils.15 In another
study, using accelerated storage tests to investigate commercial fat samples, it was
found to be a good predictor of storage stability.16

> Read full chapter

Sensory Evaluation of Oils/Fats and
Oil/Fat–Based Foods
Xiaoqing Yang, Robin A. Boyle, in Oxidative Stability and Shelf Life of Foods Con-
taining Oils and Fats, 2016

3.5.4 Anisidine Value

The anisidine value (AV) method measures the secondary oxidation compounds,
primarily 2-alkenals and 2,4-alkadienals generated due to hydroperoxide decom-
position, and it is more sensitive to unsaturated aldehydes (Gordon, 2001). Since
AV represents the content of secondary oxidation products, it is used instead of, or
together with, PV to evaluate fats and oils. For example, AV is considered to be a
more reliable indicator of oxidative rancidity than PV in a long-term storage stability
study of palm-olein oil (van der Merwe et al., 2004).

However, this does not mean AV is a better chemical marker than PV. For exam-
ple, in a study to evaluate the oxidative stability of omega-3–enriched fat spreads
(oil-in-water-in-oil emulsions), PV showed fair correlations with the sensory quality
of the samples while AV showed no correlations (O’Dwyer et al., 2013). In another
study to evaluate the shelf life of oatcakes, good correlations were found between
PV and several sensory parameters, such as aroma, flavor, and aftertaste of the
oatcakes (0.78, 0.75, and 0.72, respectively), while AV did not correlate with any of
these sensory parameters (Cognat et al., 2014). One hypothesis to explain this was
that nonvolatile aldehydes measured by the AV method were converted into volatiles
through the production of additional peroxides; it was also proposed that the n-3
polyunsaturated fatty acid content in oatcake is very low (on average, 4.3 mg/g of oil
extracted); therefore, the aldehyde level that can be measured by the AV method was
not sufficient to reflect the rancidity of oatcakes (Cognat et al., 2014).

Storage condition (temperature) also tends to have an influence on using AV as

a sensory quality indicator. When determining the effects of various oils on the
chemical and sensory stability of tortilla chips under a Schaal oven and practical
storage conditions, the PV and AV of oils extracted from chips tended to support
sensory data of Schaal oven–stored chips; however, they did not support that of
the practical stored chips (Cognat et al., 2014). This difference is probably due to
the altered oxidation pathways or reaction rates of lipids under different storage
conditions, but the mechanism remains unknown.

These observations suggested that although AV measures the secondary oxidation

products, its low specificity and tendency to be influenced by other factors, such as
the physical states of lipids, fatty acid compositions, and storage conditions, greatly
limit its application as a good chemical indicator to evaluate the sensory quality of
foods.

> Read full chapter

The Stability and Shelf Life of Fats and

Oils
G. Talbot, in The Stability and Shelf Life of Food (Second Edition), 2016

16.4.1.4 Other Methods

The PV, AV, and totox value are the most commonly used tests for oxidative status
of an oil but in addition to these are some other tests worth briefly noting that are
occasionally used.

• The thiobarbituric acid (TBA) (Kishida et al., 1993) test is also used to measure
the amount of aldehydes present in the oil but, unlike the AV, the test can be
carried out on foods without the need to extract the oil.
• The Kreis test is a colorimetric test involving the reaction of phloroglucinol
with oxidized fat to give a red color. A Lovibond colorimeter (often used to
measure the colors of oils themselves) is used to assess the intensity of the
red color. A colorimetric method to measure total carbonyl content using
2,4-dinitrophenylhydrazine (DNPH) is also used to determine the degree of
hydroperoxide breakdown products present in an oil.
• Electron paramagnetic resonance (EPR) or electron spin resonance (ESR) has
also been developed as an alternative to PV measurement, particularly for
extra virgin olive oils (Ranguelova and Barr, 2013). It specifically measures
the free radicals in samples, thus making it very useful in the early stages of
propagation.
• Various other spectrometric and chromatographic methods for determining
the levels of oxidized materials in oils have been developed. Headspace gas
chromatography, for example, is particularly useful in analyzing the volatile
components produced as a result of oxidation but, by and large, these are very
specific and nonstandard methods.

> Read full chapter

The stability and shelf life of fats and
oils
G. Talbot, in Food and Beverage Stability and Shelf Life, 2011

Other methods
The peroxide value, anisidine value and totox value are the most commonly used
tests for oxidative status of an oil, but in addition to these are some other tests worth
briefly mentioning that are occasionally used.

• The thiobarbituric acid (TBA) test is also used to measure the amount of
aldehydes present in the oil but, unlike the anisidine value, the test can be
carried out on foods without the need to extract the oil.
• The Kreis test is a colorimetric test involving the reaction of phloroglucinol
with oxidised fat to give a red colour. A Lovibond colorimeter (often used to
measure the colours of oils themselves) is used to assess the intensity of the
red colour.
• Various spectrometric and chromatographic methods for determining the
levels of oxidised materials in oils have been developed. Headspace gas
chromatography, for example, is particularly useful in analysing the volatile
components produced as a result of oxidation but, by and large, these are very
specific and non-standard methods.

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Focaccia Italian Flat Fatty Bread

Antonella Pasqualone, ... Tommaso Gomes, in Flour and Breads and their Fortificat-
ion in Health and Disease Prevention, 2011

Oxidative and hydrolytic degradation

Results of analyses of peroxide value (PV) and p-anisidine value (p-AV) provided initial
information about the oxidative degradation that affects olive oil during focaccia
baking. Total oxidation (TOTOX) of the oils from focaccias, calculated as 2 PV + p-AV,
was two- or threefold greater than in the unbaked oil, with particularly high values
in focaccias topped with onions and rosemary (Table 5.2).

Table 5.2. Mean Results of the Indices of Oxidative and Hydrolytic Degradation
Determined in Each Oil Samplea
Sample Extra-Virgin Olive Oil Oil Extracted From
Potato-Topped Focaccia Tomato-Topped Fo- Onion-Topped Focac- Rosemary-Topped Fo-
caccia cia caccia
TOTOXb 38.7 78.0 85.4 125.8 127.2

PCc 3.34 ± 0.10a 3.68 ± 0.06b 5.44 ± 0.10c 4.78 ± 0.12d 5.55 ± 0.12c

TGPc 0.08 ± 0.01a 0.13 ± 0.03b 0.22 ± 0.03c 0.27 ± 0.02d 0.25 ± 0.03c

ox-TGc 0.75 ± 0.04a 0.73 ± 0.05a 1.06 ± 0.13b 1.69 ± 0.19c 2.08 ± 0.32d

DGc 1.63 ± 0.16a 1.55 ± 0.01a 1.95 ± 0.05b 1.83 ± 0.06b 1.85 ± 0.05b

2 TGP% + 0.91 ± 0.04a 0.99 ± 0.06b 1.50 ± 0.12c 2.23 ± 0.15d 2.58 ± 0.31d
ox-TG%c

DG, diglycerides; ox-TG, oxidized triglycerides; p-AV, p-anisidine value; PC, polar
compounds; PV, peroxide value; TGP, triglyceride polymers; TOTOX = 2 PV + p-AV.

a This table shows the results of analyses performed on unbaked oil and on
oil extracted from four focaccia types, after baking, to assess the degree of
oxidative and hydrolytic degradation.
b Mean values of two independent repetitions.

c Results of statistical analysis at p < 0.05. Mean values of three independent

repetitions ± SD; one common letter following an entry indicates no signif-
icant difference.

Source: Modified with permission from Delcuratolo, D., Gomes, T., Paradiso, V. M.,
and Nasti, R. (2008). Changes in the oxidative state of extra-virgin olive oil used
in baked Italian focaccia topped with different ingredients. Food Chemistry, 106,
222–226.

High-performance size-exclusion chromatography (HPSEC) of the polar com-

pounds (PC) provided more detailed information about both the oxidative and
the hydrolytic degradation of oil via the determination of the following classes of
compounds: triglyceride oligopolymers (TGP), oxidized triglycerides (ox-TG), and
diglycerides (DG).

The HPSEC chromatograms of PC detected in the uncooked oil and in the oil
extracted from onion-topped focaccia are shown in Figure 5.4. The uncooked oil
already contained detectable amounts of TGP that were indicative of the inception of
the oxidative process, but the same oil, extracted from the baked focaccia, contained
substantially higher TGP and ox-TG levels.
Figure 5.4. HPSEC chromatograms of the polar compounds in unbaked extra-virgin
olive oil (A) and in the same oil extracted from onion-topped focaccia after baking
(B).The chromatogram shows the trend of the polar compound classes: 1, triglyc-
eride oligopolymers; 2, oxidized triglycerides; 3, diglycerides.Source: Reprinted with
permission from Delcuratolo, D., Gomes, T., Paradiso, V. M., and Nasti, R. (2008).
Changes in the oxidative state of extra-virgin olive oil used in baked Italian focaccia
topped with different ingredients. Food Chem. 106, 222–226.

The amount of PC in the oils from all the baked focaccias was significantly
greater than that in the uncooked oil. The smallest difference was observed in the
potato-topped focaccia (10% increase over the uncooked oil). In the oil from the
other three types of focaccia, increases in PC ranged from 43 to 66%.

The amount of ox-TG measured in the uncooked oil (0.75%) was not statistically
different from the amount measured in the oil extracted from the potato-topped
focaccia. By contrast, significant differences were registered in the other types of
focaccia, with ox-TG values ranging from 1.4 to 2.7 times the amount measured in
the uncooked oil. These findings confirm that the oils from the focaccias topped
with tomatoes, onions, and rosemary had undergone more intense oxidation, as
highlighted by the TOTOX values.

Baking led to the formation of oligopolymers in all the considered focaccias. The
amount of TGP in the uncooked oil was 0.08%, whereas in the potato-topped focac-
cia it was 0.13%, namely 1.5 times higher. In the other three cases, the amount of
TGP was almost threefold that of the starting oil. The evaluation of the oligopolymers
provided further evidence that the oil extracted from the potato-topped focaccia
presented a less intense degradation than the other baked oils.

The amount of DG in the uncooked oil was 1.63%. Significantly higher values were
found in the baked focaccias, except for the potato-topped type.

Table 5.2 shows the values of the sum of 2 TGP% + ox-TG%, a parameter that
provides a better evaluation of the overall oxidation (Gomes et al., 2003). Substantial
increases in overall oxidative degradation were found in oils extracted from focaccias
topped with tomato, onion, or rosemary. Again, the oil from the potato-topped
focaccia seemed to be less affected by the baking process, with an overall oxidation
index (2 TGP% + ox-TG%) of 0.99 compared to 0.91 for the uncooked oil. As already
shown by the TOTOX index, the oils from the focaccias topped with onions and
rosemary proved to be the most oxidized. TOTOX and (2 TGP% + ox-TG%) for all
samples showed a substantial agreement and a positive correlation (p < 0.05).

> Read full chapter

Chemical and physical deterioration of

bulk oils and shortenings, spreads and
frying oils
F.D. Gunstone, S. Martini, in Chemical Deterioration and Physical Instability of Food
and Beverages, 2010

14.3.2 Changes occurring during frying

Frying oils are monitored by measuring properties such as: acid value, peroxide
value, anisidine value, conjugated diene, total polar materials (TPM), and polymeric
triglycerides (PTG). The oil should be replaced when these parameters are above
specification. It has been recommended that TPM should not exceed 24% and PTG
should not exceed 12%. In most European countries, frying oil should be discarded
when TPM and PTG together exceed 24–27% (Gertz, 2006). It would be better if these
standards could be made more uniform. These measurements require laboratory
procedures and a number of simple instrumental methods suitable for use in a
food-processing factory have also been examined. It must be remembered that
oxidative deterioration starting during food processing can continue during storage
even at − 5 to + 10 °C.

The level of polar compounds is determined through elution of non-polar material

from a silica column using toluene and polymeric triacylglycerols are measured by
high-performance liquid chromatography (HPLC) of the frying oil in tetrahydrofuran
solution using a refractive index detector. These procedures require a laboratory
and other simpler robust procedures that can be used in the kitchen have been
developed. Stier (2004) has reviewed tests available to monitor deep-frying fats and
oils.

As a consequence of changes occurring during frying triacylglycerols containing

octanoic and other short chain acids are produced. These can be identified and
measured by GC after methanolysis and it is suggested that this becomes the basis
of a method of detecting the onset of rancidity at an early stage (Bruhl et al., 2008;
Velaso et al., 2004).

> Read full chapter

Understanding and reducing oxidative

flavour deterioration in foods
C. Jacobsen, in Oxidation in Foods and Beverages and Antioxidant Applications:
Understanding Mechanisms of Oxidation and Antioxidant Activity, 2010

6.2.3 Correlations between sensory data and data from chemi-

cal or instrumental measurements of lipid oxidation
Measurement of lipid oxidation can be carried out by a wide range of methods such
as peroxide value (PV), anisidine value (AV), thiobarbituric acids reacting substances
(TBARS) as well as instrumental methods such as HPLC, GC-MS, NIR, FTIR and
DSC. In lipid oxidation studies on meat products there seems to be a fairly good
correlation between TBARS and sensory data, although in many cases no attempts
have been made to statistically evaluate the correlations (Eckert et al. 1997; Murano
et al. 1998; Winne and Dirinck 1997). However, newer data indicate that even in
meat, TBARS may be an unreliable method (see further discussion in Section 6.3.6
below; Summo et al. 2010).

PV and AV are often used to evaluate the lipid oxidation in oils and the author’s
experience is that AV, at least to some extent, correlates with sensory data in oxidized
oils. In contrast, findings from fish oil enriched spreads clearly showed that the
correlation between sensory and PV and AV data was very poor (Jacobsen 1999b)
(Fig. 6.1(a) and (b)). It is not surprising that PV did not correlate with sensory data
since lipid hydroperoxides are not responsible for off-flavour formation. Moreover,
since lipid hydroperoxides are broken down to secondary oxidation products during
the later stages of oxidation a low PV does not necessarily indicate that the lipids are
not oxidized. However, it was surprising that AV in some samples was high despite
a high flavour acceptability score. This finding could indicate that the sensitivity
and specificity of this method is too low to detect the changes in concentrations of
volatiles responsible for the off-flavour formation. Therefore, AV cannot be recom-
mended for measurement of lipid oxidation in complex food systems.

Fig. 6.1. (a) Peroxide values, and (b) Anisidine values versus overall flavour acceptabil-
ity obtained from storage experiment with fish oil enriched spreads.Reprinted from
Jacobsen, C. (1999b). Sensory impact of lipid oxidation in complex food systems.
Fett/Lipid 12, 484–492 with permission from Wiley.

Good correlations between sensory data and data from headspace GC analysis
on a range of different oxidized products such as boiled potatoes (Blanda et al.
2010), fish (Milo and Grosch 1995), fish oil enriched milk (Venkateshwarlu et al.
2004a), mayonnaise and dressing (Hartvigsen et al. 2000; Let et al. 2007) have been
extensively reported in the literature and headspace GC analysis can therefore be
recommended as one of the best methods to chemically assess oxidative flavour
deterioration. Different headspace methods are available for collection of the volatile
oxidation products including static headspace, dynamic headspace and solid phase
micro extraction. It is beyond the scope of this chapter to discuss these methods in
further details, but the reader may refer to Chapter 8 for more information.

> Read full chapter

Volume 2
Charlotte Jacobsen, in Encyclopedia of Food Chemistry, 2019

Analytical Methods for Lipid Oxidation Measurements

Measurement of lipid oxidation can be carried out by a wide range of methods such
as peroxide value (PV), anisidine value (AV), thiobarbituric acids reacting substances
(TBARS) as well as instrumental methods such as HPLC, GC-MS, NIR, FTIR and DSC.
Sensory evaluation of oxidative flavor deterioration is another important method
that should always be included at some stage to understand how lipid oxidation
has impacted sensory properties of the food product in question. In lipid oxidation
studies on meat products there seems to be a fairly good correlation between TBARS
and sensory data, although in many cases no attempts have been made to statistically
evaluate the correlations (Eckert et al., 1997; Murano et al., 1998; Winne and Dirinck,
1997). However, newer data indicate that even in meat TBARS may be an unreliable
method (Summo et al., 2010). This may be related to the fact mentioned above
that lipid oxidation have several alternate pathways. Therefore, if lipid oxidation is
only followed by measuring formation of TBARS and peroxide values the major
proportions of oxidation products may be missed and the extent of lipid oxidation
may be significantly underestimated. The same is the case for studies, which only
measure peroxide value and hexanal (aldehyde formed from n-6 fatty acids). In
complex foods, the situation is even more complicated. This is due to the fact that
lipid oxidation radicals and secondary oxidation products (e.g. aldehydes) react with
other components in foods, particularly proteins. This type of co-oxidation will thus
consume lipid oxidation intermediates and products and at the same time other
substrates in the food system will be affected by oxidation. If only the traditional
lipid oxidation measurements are performed, lipid oxidation can be underestimated,
when it is in fact spread to other components of the food system, which may have
an effect on sensory properties of the food product.

Good correlations data from headspace GC analysis and sensory data have been
found for a range of different oxidized products such as fish (Milo and Grosch,
1995), fish oil enriched milk (Venkateshwarlu et al., 2004), mayonnaise and dressing
(Hartvigsen et al., 2000; Let et al., 2007a) and boiled potatoes (Blanda et al., 2010).
Headspace GC analysis can therefore be recommended as one of the best methods
to chemically assess oxidative flavor deterioration. Different headspace methods are
available for collection of the volatile oxidation products including static headspace,
dynamic headspace, purge and trap and solid phase microextraction. Thomsen et al.
(2016) have recently compared the performance of some of these methods in neat
oil and two emulsified systems.

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Quality and Control
H.B.W. Patterson, in Hydrogenation of Fats and Oils (Second Edition), 2011

Specifications of Present Oxidative State and Resistance to Fur-

ther Oxidation
The specification of the processed oil may state that it shall not already be oxidized
beyond a certain level [PV and AnV (anisidine value) tests], and that it shall have
a certain minimal resistance to further oxidation (Swift, Quality, E232/E268 tests).
Side-by-side with these tests must go the realization that a minute concentration of
off-flavor precursor can cause an easily detected bad flavor following a very modest
degree of further oxidation. If absent, the possibility that considerable oxidation may
occur before the evil flavor due to rancidity is recognizable, since if due to fatty
acid alone, this may reach 0.3% w/w before becoming objectionable. Deodorization
reduces FFA% to one-tenth of this level initially.

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Measuring antioxidant activity

DrMichael H. Gordon, in Antioxidants in Food, 2001

4.3.5 Para-anisidine value

Para-anisidine is a reagent that reacts with aldehydes to give products that absorb
at 350nm (Fig. 4.2). The p-anisidine value is defined as the absorbance of a solution
resulting from the reaction of 1 g fat in isooctane solution (100ml) with p-anisidine
(0.25% in glacial acetic acid). The products formed by reaction with unsaturated alde-
hydes (2-alkenals) absorb more strongly at this wavelength, and consequently the
test is particularly sensitive to these oxidation products. Although the test does not
distinguish between volatile and non-volatile products, the palate is generally more
sensitive to unsaturated volatile aldehydes than to saturated volatile aldehydes, so
the test is a reasonable way to assess secondary oxidation products. Measurements
of p-anisidine value are commonly used together with peroxide value measurements
in describing the total extent of oxidation by the Totox value, which equals the sum
of the p-anisidine value plus twice the peroxide value. However, the Totox value is
an empirical parameter since it corresponds to the addition of two parameters with
different units.
 4.2. The reaction of p-anisidine with alkenals.

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