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UNIT F1.2
Anthocyanin pigment content has a critical role in the color quality of many fresh and
processed fruits and vegetables. Thus, accurate measurement of anthocyanins, along with
their degradation indices, is very useful to food technologists and horticulturists in
assessing the quality of raw and processed foods. Since many natural food colorants are
anthocyanin derived (e.g., grape-skin extract, red-cabbage extract, purple-carrot extract),
the same measurements can be used to assess the color quality of these food ingredients.
In addition, there is intense interest in the anthocyanin content of foods and nutraceuticals
because of possible health benefits such as reduction of coronary heart disease (Bridle
and Timberlake, 1996), improved visual acuity (Timberlake and Henry, 1988), antioxidant
activities (Takamura and Yamagami, 1994; Wang et al., 1997), and anticancer activities
(Karaivanova et al., 1990; Kamei et al., 1995). Substantial quantitative and qualitative
information can be obtained from the spectral characteristics of anthocyanins. The
protocols described in this unit rely on the structural transformation of the anthocyanin
chromophore as a function of pH, which can be measured using optical spectroscopy. The
pH-differential method, a rapid and easy procedure for the quantitation of monomeric
anthocyanins, is first described (see Basic Protocol 1). In addition, other auxiliary
spectrophotometric techniques are used to measure the extent of anthocyanin polymerization and browning (see Basic Protocol 2).
TOTAL MONOMERIC ANTHOCYANIN BY THE pH-DIFFERENTIAL
METHOD
Anthocyanin pigments undergo reversible structural transformations with a change in pH
manifested by strikingly different absorbance spectra (Fig. F1.2.1). The colored oxonium
form predominates at pH 1.0 and the colorless hemiketal form at pH 4.5 (Fig. F1.2.2).
The pH-differential method is based on this reaction, and permits accurate and rapid
measurement of the total anthocyanins, even in the presence of polymerized degraded
pigments and other interfering compounds.
BASIC
PROTOCOL 1
2.0
1.8
Absorbance
1.6
1.4
pH 1.0
1.2
1.0
0.8
0.6
0.4
0.2
0.0
260
pH 4.5
360
460
Wavelength (nm)
560
660
760
Figure F1.2.1 Spectral characteristics of purified radish anthocyanins (acylated pelargonidin-3sophoroside-5-glucoside derivatives) in pH 1.0 and pH 4.5 buffers.
Contributed by M. Mnica Giusti and Ronald E. Wrolstad
Current Protocols in Food Analytical Chemistry (2001) F1.2.1-F1.2.13
Copyright 2001 by John Wiley & Sons, Inc.
Anthocyanins
F1.2.1
R1
R1
OH
OH
O
HO
R2
H+
HO
R2
O-gly
O-gly
O-gly
O-gly
H+
+H2O
R1
H
O
R1
HO
OH
O-gly
OH
HO
chalcone: colorless
pH = 4.5
R2
O-gly
R2
O-gly
OH
O-gly
carbinol pseudo-base (hemiketal form): colorless
pH = 4.5
Materials
0.025 M potassium chloride buffer, pH 1.0 (see recipe)
0.4 M sodium acetate buffer, pH 4.5 (see recipe)
1. Turn on the spectrophotometer. Allow the instrument to warm up at least 30 min
before taking measurements.
2. Determine the appropriate dilution factor for the sample by diluting with potassium
chloride buffer, pH 1.0, until the absorbance of the sample at the vis-max (Table F1.2.1)
is within the linear range of the spectrophotometer (i.e., for most spectrophotometers
the absorbance should be less than 1.2). Divide the final volume of the sample by the
initial volume to obtain the dilution factor (DF; for example see step 7).
IMPORTANT NOTE: In order to not exceed the buffers capacity, the sample should not
exceed 20% of the total volume.
3. Zero the spectrophotometer with distilled water at all wavelengths that will be used
(vis-max and 700 nm).
Many spectrophotometers will allow for a rapid baseline correction to zero by using
baseline adjust.
Characterization
and Measurement
of Anthocyanins
by UV-Visible
Spectroscopy
4. Prepare two dilutions of the sample, one with potassium chloride buffer, pH 1.0, and
the other with sodium acetate buffer, pH 4.5, diluting each by the previously
determined dilution factor (step 2). Let these dilutions equilibrate for 15 min.
F1.2.2
Current Protocols in Food Analytical Chemistry
Table F1.2.1
Anthocyanina
Cyanidin (Cyd)
Cyd
Cyd-3-ara
Cyd-3,5-diglu
Cyd-3-gal
Cyd-3-glu
Cyd-3-rut
Cyd-3-sam-5-glu
Cyd-3-sam-5-glu + sinapic +
caffeic + malonic
Cyd-3-sam-5-glu + sinapic +
ferulic
Cyd-3-sam-5-glu + sinapic +
ferulic + malonic
Cyd-3-sam-5-glu + sinapic +
p-coum + malonic
Cyd-3-soph-5-glu
Cyd-3-soph-5-glu + malonic
Cyd-3-soph-5-glu + sinapic
Cyd-3-soph-5-glu + di-sinapic
Cyd-3-soph-5-glu + ferulic
Cyd-3-soph-5-glu + di-ferulic
Cyd-3-soph-5-glu + p-coumaric
Cyd-3-soph-5-glu +
di-p-coumaric
Delphinidin (Dpd)
Dpd
Solvent system
vis-max (nm)
Molar
Reference
absorptivity ()
510.5
547
538
24600
34700
44400
535
44460
0.1 N HCl
520
30175
Methanolic HCl
508.5
35000
530
34300
535
44900
535
46200
535
46230
HCl in methanol
Aqueous buffer, pH 1
0.1 N HCl
1% HCl in methanol
530
510
520
530
30200
26900
25740
34300
512
510
523
522
538
18800
7000
28840
3600
21200
Schou, 1927
Ribereau-Gayon, 1959
Zapsalis and Francis,
1965
Fuleki and Francis,
1968a
Niketic-Aleksic and
Hrazdina, 1972
Brouillard and El Hache
Chahine, 1980
Siegelman and
Hendricks, 1958
Sakamura and Francis,
1961
Zapsalis and Francis,
1965
Fuleki and Francis,
1968a
Swain, 1965
Jurd and Asen, 1966
McClure, 1967
Siegelman and
Hendricks, 1958
Heredia et al., 1998
Figueiredo et al., 1996
Swain, 1965
Figueiredo et al., 1996
Figueiredo et al., 1996
528
15100
538
20100
536
19000
Methanolic HCl
Methanolic HCl
Methanolic HCl
Methanolic HCl
Methanolic HCl
Methanolic HCl
Methanolic HCl
Methanolic HCl
524
528
528
530
528
530
526
528
37150
32360
37150
38020
32360
34670
38020
32360
522.5
34700
Schou, 1927
continued
F1.2.3
Current Protocols in Food Analytical Chemistry
Table F1.2.1
Anthocyanina
Solvent system
vis-max (nm)
Molar
Reference
absorptivity ()
Dpd-3-glu
1% HCl in methanol
10% ethanol, pH 1.5
543
520
29000
23700
520
557
519
545
520
37200
36200
10700
10300
37700
546
538
13900
29500
0.1 N HCl
520
28000
Methanol, pH 1.0
10% ethanol, pH 1.5
0.1% HCl in methanol
535
520
536
36400
20200
30200
Schou, 1927
Ribereau-Gayon, 1959
Schou, 1927
Ribereau-Gayon, 1959
Niketic-Aleksic and
Hrazdina, 1972
Somers, 1966
Koeppen and Basson,
1966
Niketic-Aleksic and
Hrazdina, 1972
Metivier et al., 1980
Heredia et al., 1998
Koeppen and Basson,
1966
504.5
505
17800
18420
Schou, 1927
Giusti et al., 1999
524
510
512
496
19780
32360
28000
27300
513
516
496
36600
22390
31620
15600
508
504
17330
32080
511
498
497
39591
1800020000
25370
506
506
30690
24140
507
498
29636
18000-20000
506
28720
508
34889
Malvidin (Mvd)
Mvd
Mvd-3,5-diglu
Mvd-3-glu
Mvd-3-glu + p-coum
Pelargonidin (Pg)
Pg
Pg-3,5-diglu
Pg-3-(dicaffeoylglu)-soph-5-glu
Pg-3-glu
Pg-3-rut-5-glu + p-coumaric
Pg-3-soph-5-glu
Pg-3-soph-5-glu + ferulic
Pg-3-soph-5-glu caffeoyl
derivatives
Pg-3-soph-5-glu + p-coumaric
1% HCl
1% HCl in ethanol
0.025 M potassium
chloride buffer, pH 1.0
0.1% HCl in methanol
0.025 M potassium
chloride buffer, pH 1.0
0.1% HCl in methanol
Aqueous buffer, pH 0.8
0.025 M potassium
chloride buffer, pH 1.0
0.1% HCl in methanol
0.025 M potassium
chloride buffer, pH 1.0
0.1% HCl in methanol
Aqueous buffer, pH 0.8
0.025 M potassium
chloride buffer, pH 1.0
0.1% HCl in methanol
continued
F1.2.4
Current Protocols in Food Analytical Chemistry
Table F1.2.1
Anthocyanina
Solvent system
vis-max (nm)
Molar
Reference
absorptivity ()
508
33010
508
508
39785
31090
508
39384
511
532
37200
40800
532
46100
532
46070
Pnd-3,5-diglu
0.1 N HCl
520
36654
Pnd-3-gal
532
48400
532
48400
531
48340
536
512
11300
14100
Schou, 1927
Sakamura and Francis,
1961
Zapsalis and Francis,
1965
Fuleki and Francis,
1968a
Niketic-Aleksic and
Hrazdina, 1972
Sakamura and Francis,
1961
Zapsalis and Francis,
1965
Fuleki and Francis,
1968a
Somers, 1966
Heredia et al., 1998
0.1 N HCl
520
33040
HCl in methanol
0.1% HCl in methanol
10% ethanol, pH 1.5
535
546
520
23440
12900
18900
Pnd-3-glu
Petunidin (Ptd)
Ptd-3,5-diglu
Ptd-3-glu
Niketic-Aleksic and
Hrazdina, 1972
Swain, 1965
Somers, 1966
Heredia et al., 1998
aAbbreviations: ara: arabinoside; gal: galactoside; glu: glucoside; rut: rutinoside; sam: sambubioside; soph: sophoroside.
5. Measure the absorbance of each dilution at the vis-max and at 700 nm (to correct for
haze), against a blank cell filled with distilled water.
All measurements should be made between 15 min and 1 hr after sample preparation, since
longer standing times tend to increase observed readings.
Absorbance readings are made against water blanks, even if the samples are in buffer or
bisulfite solutions, as buffer or bisulfite absorbance is nil at the measured wavelengths.
The authors have compared the values obtained by using water as a blank as compared
with buffer or bisulfite as blanks in different systems and have found no difference in the
final values obtained for monomeric and/or polymeric anthocyanin content; on the other
hand, reading the diluted samples against the corresponding buffer and/or bisulfite solution
is more time-consuming and extends the procedure unnecessarily.
The samples to be measured should be clear and contain no haze or sediments; however,
some colloidal materials may be suspended in the sample, causing scattering of light and
a cloudy appearance (haze). This scattering of light needs to be corrected for by reading
at a wavelength where no absorbance of the sample occurs, i.e., 700 nm.
Anthocyanins
F1.2.5
Current Protocols in Food Analytical Chemistry
Table F1.2.2
in Naturea
Anthocyanidins
Pelargonidin
Petunidin
Malvidin
Hex
Hex H2Ob
Acd + 1 hex
Acd + 2 hex
Acd + 3 hex
271
180.2
162.2
433.2
595.4
757.6
287
180.2
162.2
449.2
611.4
773.6
301
180.2
162.2
463.2
625.4
787.6
303
180.2
162.2
465.2
627.4
789.6
317
180.2
162.2
479.2
641.4
803.6
331
180.2
162.2
493.2
655.4
817.6
Pent
Pent H2Ob
Acd + 1 pent
Acd + 1 hex + 1 pent
150.0
132.0
403.0
565.2
150.0
132.0
419.0
581.2
150.0
132.0
433.0
595.2
150.0
132.0
435.0
597.2
150.0
132.0
449.0
611.2
150.0
132.0
463.0
625.2
Rhamnose
Rutinose
Rutinose H2Ob
Acd + rutinose
Acd + rutinose + 1 hex
Acd + rutinose + 1 pent
164.2
326.2
308.2
579.2
741.4
711.2
164.2
326.2
308.2
595.2
757.4
727.2
164.2
326.2
308.2
609.2
771.4
741.2
164.2
326.2
308.2
611.2
773.4
743.2
164.2
326.2
308.2
625.2
787.4
757.2
164.2
326.2
308.2
639.2
801.4
771.2
164.2
180.2
194.2
224
82
96.1
104.1
118.1
146.2
162.2
176.2
206
64
78.1
86.1
100.1
Characterization
and Measurement
of Anthocyanins
by UV-Visible
Spectroscopy
IMPORTANT NOTE: The MW and used in this formula correspond to the predominant
anthocyanin in the sample. Use the reported in the literature for the anthocyanin pigment
in acidic aqueous solvent. If the of the major pigment is not available, or if the sample
composition is unknown, calculate pigment content as cyanidin-3-glucoside, where MW =
449.2 and = 26,900 (see Background Information, discussion of Molar Absorptivity).
The equation presented above assumes a pathlength of 1 cm.
F1.2.6
Current Protocols in Food Analytical Chemistry
BASIC
PROTOCOL 2
Indices for anthocyanin degradation of an aqueous extract, juice, or wine can be derived
from a few absorbance readings of a sample that has been treated with sodium bisulfite.
Anthocyanin pigments will combine with bisulfite to form a colorless sulfonic acid adduct
(Figure F1.2.3). Polymerized colored anthocyanin-tannin complexes are resistant to
bleaching by bisulfite, whereas the bleaching reaction of monomeric anthocyanins will
rapidly go to completion. The absorbance at 420 nm of the bisulfite-treated sample serves
as an index for browning. Color density is defined as the sum of absorbances at the vis-max
and at 420 nm. The ratio between polymerized color and color density is used to determine
the percentage of the color that is contributed by polymerized material. The ratio between
monomeric and total anthocyanin can be used to determine a degradation index.
Materials
Bisulfite solution (see recipe)
0.025 M potassium chloride buffer, pH 1.0 (see recipe)
1. Turn on the spectrophotometer and allow the instrument to warm up at least 30 min
before taking measurements.
2. Determine the appropriate dilution factor for the sample by diluting with 0.025 M
potassium chloride buffer, pH 1.0 until the absorbance of the sample at the vis-max is
within the linear range of the spectrophotometer (i.e., for most spectrophotometers
the absorbance should be less than 1.2). Divide the final volume of the sample by the
initial volume to obtain the dilution factor (DF; for example see step 6).
3. Zero the spectrophotometer with distilled water at all wavelengths that will be used
(420 nm, vis-max, 700 nm).
Many spectrophotometers will allow for a rapid baseline correction to zero by using
baseline adjust.
4. Dilute the sample with distilled water using the dilution factor already determined
(step 2). Transfer 2.8 ml of the diluted sample to each of two cuvettes. Add 0.2 ml of
bisulfite solution to one and 0.2 ml distilled water to the other. Equilibrate for 15 min.
It is critical that the pH not be adjusted to highly acidic conditions (e.g., pH 1) but rather
be in the typical pH range of fruit juices and wines, or higher (e.g., pH 3). Highly acidic
conditions will reverse the bisulfite addition reaction and render the measurement invalid.
5. Measure the absorbance of both samples at 420 nm, vis-max, and 700 nm (to correct
for haze), against a blank cell filled with distilled water.
All measurements should be made between 15 min (see step 4) and 1 hr after sample
preparation and bisulfite treatment. Longer standing times tend to increase observed
readings.
Absorbance readings are made against water blanks, even if the samples are in buffer or
bisulfite solutions, as buffer or bisulfite absorbance is nil at the measured wavelengths.
The authors have compared the values obtained by using water as a blank as compared
with the use buffer or bisulfite as a blank in different systems and have found no difference
in the final values obtained for monomeric and/or polymeric anthocyanin content; on the
other hand, reading the samples against the corresponding buffer and/or bisulfite solution
is more time-consuming and extends the procedure unnecessarily.
The samples to be measured should be clear and contain no haze or sediments; however,
some colloidal materials may be suspended in the sample, causing scattering of light and
a cloudy appearance (haze). This scattering of light needs to be accounted for by reading
at a wavelength where no absorbance of the sample occurs (i.e., 700 nm).
Anthocyanins
F1.2.7
Current Protocols in Food Analytical Chemistry
6. Calculate the color density of the control sample (treated with water) as follows:
Color density = [(A420 nm A700nm) + (A vis-max A700 nm)] DF
where DF is the dilution factor (for example, if 0.2 ml sample diluted to 3 ml, DF =
15)
7. Calculate the polymeric color of the bisulfite bleached sample as follows:
Polymeric color = [(A420 nm A700 nm) + (A vis-max A700 nm)] DF
8. Calculate the percent polymeric color using the formula:
Percent polymeric color = (polymeric color/color density) 100
REAGENTS AND SOLUTIONS
Use deionized or distilled water in all recipes and protocol steps. For common stock solutions, see
APPENDIX 2A; for suppliers, see SUPPLIERS APPENDIX.
Bisulfite solution
Dissolve 1 g of potassium metabisulfite (K2S2O5) in 5 ml of distilled water.
This reagent must be prepared the same day as the readings; otherwise, it develops a yellow
color that will contribute to the absorbance readings and interfere with the quantitation.
COMMENTARY
Background Information
Characterization
and Measurement
of Anthocyanins
by UV-Visible
Spectroscopy
1994; Wang et al., 1997) and anticancer properties (Karaivanova et al., 1990; Kamei et al.,
1995). Anthocyanins have also found considerable potential in the food industry as safe and
effective food colorants (Strack and Wray,
1994); interest in this application has increased
in recent years. In 1980, the annual world production had been estimated as reaching 10,000
tons from grapes alone (Timberlake, 1980).
Quantitative and qualitative anthocyanin composition are important factors in determining
the feasibility of the use of new plant materials
as anthocyanin-based colorant sources.
Frequently, it is desirable to express anthocyanin determinations in terms that can be compared with the results from different workers.
The best way to express these results is in terms
F1.2.8
Current Protocols in Food Analytical Chemistry
R1
R1
OH
OH
O
HO
HO
R2
O-gly
OH
Strong acid
R2
O-gly
OH SO3H
bisulfite addition compound: colorless
will decolor anthocyanins but not affect interfering materials. A measurement of the absorbance at the visible maximum is obtained, followed by bleaching and remeasuring to give a
blank reading (Jackman et al., 1987). The two
most used bleaching agents are sodium sulfite
(Somers and Evans, 1974; Wrolstad et al.,
1982) and hydrogen peroxide (Swain and Hillis, 1959).
By using both of these spectral procedures,
accurate measurement of the total monomeric
anthocyanin pigment content can be obtained,
along with indices for polymeric color, color
density, browning, and degradation. To determine total anthocyanin content, the absorbance
at pH 1.0 and 4.5 is measured at the vis-max
(Table F1.2.1) and at 700 nm, which allows for
haze correction. The bisulfite bleaching reaction is utilized to generate the various degradation indices. While monomeric anthocyanins
are readily bleached by bisulfite at product pH
(Fig. F1.2.3), the polymeric anthocyanin-tannin and melanoidin pigments are resistant and
will remain colored. Somers and Evans (1974)
used this reaction in developing spectral methods for assessing the color quality of wines. The
authors laboratory has found them useful for
tracking color quality in a wide range of anthocyanin-containing foods (Wrolstad et al., 1982,
1995). Absorbance measurements are taken at
the vis-max and at 420 nm on the bisulfite
bleached and control samples. Color density is
the sum of the absorbances at the vis-max and
at 420 nm of the control sample, while polymeric color is the same measurement for the
bisulfite treated sample. A measure of percent
polymeric color is obtained as the ratio between
these two indexes. The absorbance at 420 nm
of the bisulfite-treated sample is an index for
browning, as the accumulation of brownish
Anthocyanins
F1.2.9
Current Protocols in Food Analytical Chemistry
Characterization
and Measurement
of Anthocyanins
by UV-Visible
Spectroscopy
Spectral characteristics
Substantial information can be obtained
from the spectral characteristics of anthocyanins (Fig. F1.2.1). Two distinctive bands of absorption, one in the UV-region (260 to 280 nm)
and another in the visible region (490 to 550
nm) are shown by all anthocyanins. The different aglycons have different vis-max, ranging
from 520 nm for pelargonidin to 546 nm for
delphinidin, and their monoglucosides exhibit
their vis-max at about 10 to 15 nm lower (Strack
and Wray, 1989). The shape of the spectrum
may give information regarding the number
and position of glycosidic substitutions and
number of cinnamic acid acylations. The ratio
between the absorbance at 440 nm and the
absorbance at the vis-max is almost twice as
much for anthocyanins with glycosidic substitutions in position 3 as compared to those with
substitutions in positions 3 and 5 or position 5
only. The presence of glycosidic substitutions
at other positions (e.g., 3,7-diglycosides) can
be recognized because they exhibit a different
spectral curve from those of anthocyanins with
common substitution patterns. The presence of
cinnamic acid acylation is revealed by the presence of a third absorption band in the 310 to
360 nm range (Figure F1.2.1), and the ratio of
absorbance at 310 to 360 nm to the absorbance
at the visible vis-max will give an estimation of
the number of acylating groups (Harborne,
1967; Hong and Wrolstad, 1990). The solvent
used for spectral determination will affect the
position of the absorption bands, and therefore
must be taken into consideration when comparing available data.
F1.2.10
Current Protocols in Food Analytical Chemistry
Table F1.2.3
Source
Apples (Scugog)
Bilberries
Blackberries
Black currants
Blueberries
Red cabbage
Black chokeberries
Cherries
Cranberries
Elderberry
Grapes
Kiwi
Red onions
Plum
Red radishes
Black raspberries
Red Raspberries
Strawberries
Tradescantia pallida
(leaves)
Pigment content
Reference
(mg/100 g fresh weight)
10
300320
83326
130400
25495
25
560
4450
60200
450
6600
100
721
225
1160
300400
2060
1535
120
a density of 1 g/ml for juice). Then, the appropriate dilution factor for the sample was determined by diluting 0.2 ml of the solution with
2.8 ml of 0.025 M potassium chloride buffer,
pH 1.0 (dilution factor = 15). To calculate
monomeric anthocyanin content, color density,
or polymeric color, the dilution factor to use
would be: DF = (10 15) = 150.
The methodologies used to measure color
density and polymeric color were developed for
fruit juices, which naturally have an acidic pH.
If the material to be measured has a pH in the
neutral or alkaline range, the pH of the solution
should be lowered with a weak acid. In these
cases, the authors recommend the use of a 0.1
M citric acid buffer, pH 3.5, instead of distilled
water to prepare the different dilutions.
Some potential interfering materials are
other red pigments: FD&C Red No. 40, FD&C
Red No. 3, cochineal, and beet powder (betalain
pigments). The presence of alternative colorants may be suspected if the vis-max at pH 1.0
is high (550 nm, more typical of betalain pigments), or if a bright red coloration is found at
pH 4.5 (potential presence of artificial dyes).
The presence of ethanol does not interfere
with the assay at the levels typically encountered in wines (10% to 14%).
Highly acylated anthocyanins may not respond to pH changes the same way as anthocyanins with no or few acylating groups, and may
not decolor as much as nonacylated or monoor diacylated anthocyanins do at pH 4.5.
Anticipated Results
The anthocyanin content of different common fruits and vegetables is presented in Table
F1.2.3. Anthocyanin-containing fruit or vegetable juices typically have pigment content
ranging from 50 to 500 mg/liter. Anthocyaninbased natural colorants and nutraceuticals may
have a much higher pigment concentration, on
the order of a few grams/liter.
Fresh fruit or vegetable juices should have
a low percentage of polymeric color (usually
less than 10%), while processed samples and
materials subjected to storage abuse will be
much higher (30% or more). This is highly
variable, dependent on the commodity, processing conditions, and storage history.
Always express anthocyanin pigment content in terms of the specific anthocyanin used
for calculation, and specify molecular weight
and utilized.
Anthocyanins
F1.2.11
Current Protocols in Food Analytical Chemistry
Time Considerations
Quantitation of anthocyanins can be
achieved in <1 hr. It is necessary to wait for the
spectrophotometer to warm up, and for the
diluted samples to equilibrate at least 15 min.
The absorbance readings take a few minutes.
Literature Cited
Asen, S., Stuart, N.W., and Siegelman, H.W. 1959.
Effect of various concentrations of nitrogen,
phosphorus and potassium on sepal color of Hydrangea macrophylla. Am. Soc. Hort. Sci.
73:495-502.
Bridle, P. and Timberlake, C.F. 1996. Anthocyanins
as natural food colors-selected aspects. Food
Chem. 58:103-109.
Brouillard, R. and El Hache Chahine, J.M. 1980.
Chemistry of anthocyanin pigments. 6. Kinetic
and thermodynamic study of hydrogen sulfite
addition to cyanin. Formation of a highly stable
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Characterization
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Spectral methods are described for generating several color quality indices for wines.
Description of the pH differential method for determination of total anthocyanins and indices for anthocyanin degradation as applied to fruit juices and
wines.
Anthocyanins
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Current Protocols in Food Analytical Chemistry