ANALYTICAL SCIENCES OCTOBER 1998, VOL.

14 889
1998 © The Japan Society for Analytical Chemistry

Reviews

Photometric Methods for the Determination of Vitamin C

Satya P. ARYA†, Meenakshi MAHAJAN and Preeti JAIN

Department of Chemistry, Kurukshetra University, Kurukshetra–136119, Haryana State, India

The importance of vitamin C to the human body is widely acknowledged throughout the globe. The deficiency of this
vitamin leads to various diseases. In view of its importance, numerous methods including spectrophotometric ones have
been developed for its determination in pharmaceuticals, foods and biological samples. A comprehensive review of the
available spectophotometric methods for the determination of ascorbic acid is presented.

Keywords Vitamin C determination, spectrophotometric method

ed through the Chemical Abstracts for the period 1970

1 Introduction to mid-1997.

Vitamin C occurs in different concentrations in a vari-

ety of natural samples. It is added to several pharma-
ceutical products as an essential ingredient, a stabilizer
for vitamin B complex, and as an antioxidant.
Consequent upon its desirable effects, it is widely used
in the treatment of certain diseases such as scurvy,
common cold, anemia, haemorrhagic disorders, wound
healing, and even infertility, to mention some stark
cases. It is considered essential for the development
and regeneration of muscles, bones, teeth and skin.
The increasing use of pharmaceuticals and other natural
samples containing vitamin C has meant that the practi-
cising chemists should develop analytical procedures
for its determination which are simple to operate, rapid,
accurate, sensitive and selective. The desire to develop
methods with ideal characteristics has resulted a large
number of procedures with varying applicability. Many
instrument-based analyses including fluorometry 1–4,
HPLC5–10, polarography11–13 and enzymatic14,15 methods
are reported in the literature. But due to their inherent
limitations, these techniques are not commonly used for
routine analyses. However, photometric methods are
particularly attractive because of their speed and sim-
plicity. Consequently, a large number of such proce-
dures have been developed for the determination of
ascorbic acid (AA). Though some short reviews16–18
have been reported, a critical assessment of these meth-
ods is desirable to examine their salient features and
utility. This review is an attempt to assess exclusively
the existing spectrophotometric methods for the deter-
mination of vitamin C as regards their simplicity, rapid-
ity, Beer’s law range, sensitivity, selectivity and applic-
ability. It is primarily based on the information collect-
2 Results and Discussion
Several dyes such as 2,6-dichlorophenolindophenol
(DCIP), dimethoxydiquinone (DMDQ), ninhydrin, fast
red AL salt and 2′,7′-dichlorofluorescein etc. have been
used for the determination of vitamin C. Among these
dyes, DCIP has been most extensively studied. It is
included in the official titrimetric methods as reported
in different pharmacopoeias19–21 and it also forms the
basis of many colorimetric methods. The blue dye
DCIP is reduced to the colorless form on addition of
ascorbic acid as shown in Fig. 1, but it gives a pink
color to the acidic solutions. Using the dye, ascorbic
acid present in human urine22 and processed potatoes23
has been determined. The excess dye can be extracted
with xylene or butanol.24 Many substances which are
capable of reducing the dye resulting from the prepara-
tion and processing of food samples interfere. Flow
dialysis proposed by Gary et al.25 and continuous flow
systems have been used to monitor the decrease in
absorbance of DCIP. Such automated systems appear
to be justified only when routine analysis of a large
Ascorbic acid DCIP Dehydroascorbic DCIP
(Oxidized, Blue-Pink) acid (Reduced, Colorless)

† To whom correspondence should be addressed. Fig. 1 The reduction of DCIP with ascorbic acid.
890
‘Indigoid’
DMDQ quinhydrone
Fig. 2 Reduction reaction of dimethoxydiquinone (DMDQ).
number of samples is needed; otherwise it is tedious to
use for a single estimation.
Dimethoxydiquinone26 gives a violet-colored product
with ascorbic acid in a phosphate buffer (pH 6.6). The
reduced “indigoid” quinhydrone form is perhaps
responsible for the formation of violet-colored product
as shown in Fig. 2. After diluting with dioxane,
absorbance of the colored solution which is stable over
24 h only under dark conditions is measured at 510 nm.
Heating leads to a decrease in color intensity. Beer’s
law holds good up to 80 µg ml–1 with a detection limit
of 10 µg ml–1. Riboflavin and copper interfere. The
interference of iron(II) sulfate responsible for precipita-
tion can be removed by centrifugation. Though the
method is not sufficiently sensitive (ε=1.62×103), it can
still be applied to the analysis of citrus fruits27 after
extracting the colored product into chloroform
(λ max=530 nm). Lin et al.28 and Pandey29 reported pro-
cedures based on the reaction of ascorbic acid with fast
Red AL salt (1) (zinc chloride salt of diazotized 1-
aminoanthraquinone) and tetrachlorobenzoquinone (2).
The reaction of (1) proceeds in acid medium but the
blue color develops only after the addition of alkali,
which exhibits three absorption bands between 500 –
630 nm. If one uses the latter reagent (2), ascorbic acid
is determined at 336 nm ( ε =535 cm 2 mol –1 ) via a
decrease in absorbance of 7×10–4 M tetrachlorobenzo-
quinone (chloranil) in 80% acetone–water (v/v) medi-
um. With these methods, mixtures of ascorbic acid
with thiols like o-mercaptobenzoic acid, mercaptosuc-
cinic acid, 3-mercaptopropionic acid can not be
resolved.
Methylene Blue 30, (3) and ninhydrin 31,32 (4) find
applications with the determination of ascorbic acid in
food products. The colorless form of the dye (3) is
extracted into chloroform after its reduction with ascor-
bic acid; back oxidation of the dihydro derivative to
Methylene Blue has been used for the assay of ascorbic
acid ( λ max=653 nm). The method is reported to be
highly sensitive. The reaction of ascorbic acid with
ninhydrin carried out on a boiling water bath using
80% aqueous solution as a medium in 0.01 M NH4OH
is used for its determination in pharmaceuticals
(λ max=415 nm), but without added advantages.
In the sixties, many methods based on the coupling of
ascorbic acid with aniline diazonium salts were report-
ed. A purplish or blue colored species is produced by
these salts with ascorbic acid in alkaline medium.
Diazotized-4-methoxy-2-nitroaniline couples with
ANALYTICAL SCIENCES OCTOBER 1998, VOL. 14
ascorbic acid in oxalic acid medium in the presence of
ethanol or isopropanol, giving a purplish color in alka-
line solutions. Though Fe(II), Sn(II) and dehydroascor-
bic acid (DHAA) do not interfere, the presence of
reductones and reductic acid requires formaldehyde
condensation. Low contents of vitamin C in the pres-
ence of flavanoids and pectic substances are also
detected. The reaction of ascorbic acid with 4-
nitrobenzene diazonium fluoroborate in acetic acid
medium was used for its determination at λ max 415 nm.
But the mixture has to be kept for 25 min in the dark,
followed by the addition of sodium hydroxide. The
sensitivity of a large number of stabilized diazonium
salts was evaluated; diazotized 4-nitroaniline-2:5-
dimethoxy-aniline was found to give the most intense
color reaction.
Enzymic33–35 colorimetric determinations of ascorbic
acid in commercial vitamin C tablets and in fruits and
vegetables were made by measuring the absorbance at
358 nm or 320 nm of the resulting products obtained by
oxidation of o-phenylenediamine/1,4-diaminobenzene
using ascorbate oxidase or peroxidase in presence of
H2O2 at pH 5.3. Ascorbic acid is determined after oxi-
dation with mercuric chloride and condensing the
DHAA with 4,5-disubstituted phenylenediamine 36,
which gives the quinoxaline derivative used for
absorbance measurement. The method involving 4-
nitro-1,2-phenylenediamine37 (λ max=375 nm) is very
complex and laborious, since it involves many time-
consuming steps including purification of the sample
with anionic Sephadex column.
In the recent past, the determination of oxidized and
reduced vitamin C in pharmaceuticals, foods and bio-
logical samples has gained importance since AA and
DHAA redox couple is an important component of
many biological systems. Simultaneous measurement
of both AA and DHAA using HPLC has been carried
out by various workers38–45 in different laboratories.
Rose and Nahrwold38 determined AA and DHAA by
monitoring UV absorbance at 254 nm and 210 nm
respectively for the analysis of foods, biological sam-
ples and pharmaceutical preparations. Graham and
Donald39 have carried out the analysis at 254 nm after
extracting the food samples with 62.5 mM metaphos-
phoric acid using an ion exchange column (Aminex-
HPX 87H). Both these forms have also been deter-
mined in vegetable samples40 using a UV detector (254
nm). Yasui and Hayashi41 made such determinations by
converting to compounds having λ max at 300 nm under
alkaline conditions. Derivatization of DHAA is accel-
erated in the presence of sodium borohydride.
Validation of the micromethod for the determination of
the oxidized and reduced vitamin C in plasma by
HPLC fluorescence method has been reported by
Tessier et al.45 These methods are useful and a single
step HPLC assay of such detections has been helpful in
overcoming the burden of derivatization.
Ascorbic acid gives colored species with substituted
benzene such as m-dinitrobenzene46 in formaldehyde
ANALYTICAL SCIENCES OCTOBER 1998, VOL. 14
and trinitrobenzene47 in tartrate buffer when studied for
its determination over the concentration ranges 2 – 50
and 0 – 125 µg ml –1 of ascorbic acid respectively.
Methanolic solution of resorcinol48 gives a pale yellow
color (λ max=425 nm) with ascorbic acid in hydrochloric
acid medium, obeying Beer’s law for 80 – 400 µg ml–1.
4-Chloro-7-nitrobenzofurazane 49 forms a bluish green
colored species with ascorbic acid in presence of 0.2 M
sodium hydroxide. The absorbance is measured at 582
nm after diluting the reaction contents with 50% (v/v)
aqueous acetone solution. Beer’s law is obeyed in the
concentration range 5 – 20 µg ml–1. The colored prod-
uct is stable for 30 min only when kept away from
direct sunlight or artificial day light. The method is
reported free from the interference of all other vitamins
and minerals present in multivitamin preparations and
can be applied to the analysis of pharmaceuticals, fresh
fruit juices and vegetables.
Hashmi et al.50 proposed a method based on the reac-
tion of 2,3,5-triphenyltetrazolium chloride with ascor-
bic acid in alkaline medium. The pink solution is
allowed to stand in the dark for 30 min at 25˚C; it
obeys Beer’s law over the range 5 – 25 µg ml–1. Sugars
(>15 µg ml–1) except sucrose interfere by forming a
similar color to that of the reagent. Riboflavin,
cyanocobalamin and folic acid interfere due to their
own color. Beutler et al.51,52 investigated the use of
methylthiazolyltetrazolium salt in presence of ascorbate
oxidase enzyme and 3-(4,5-dimethylthiazolyl-2-yl)-2,5-
diphenyltetrazolium chloride or bromide in the pres-
ence of 5-methylphenazinium methyl sulfate (electron
carrier) at pH 3.5 for the determination of ascorbic acid
in foods, fruit juices and vegetables juices. These reac-
tions involve the formation of formazon (λ max=578
nm). The interference of sulfur dioxide requires treat-
ment with formaldehyde, and color interference from
dark juices is removed by decolorization with 1%
polyvinylpolypyrrolidone before filtration. Sorbitol,
alcohol and oxalate interfere with the ascorbic acid oxi-
dase. However, the effect of oxalate can be checked by
adding a slight excess of Ca(II) ions. Other derivatives
such as 2,5-diphenyl-3-thiazolyl tetrazolium chloride53
at pH 12.2, 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-
phenyltetrazolium chloride at pH 10.5 (λ max=540 nm)
and 2,2′,5,5′-tetra-(4-nitrophenyl)-3,3′-(3,3′-dimethoxy-
4,4′-biphenyl)ditetrazolium chloride54 have also been
employed for the assay of ascorbic acid.
The coupling of 2,4-dinitrophenylhydrazine (DNPH)
with ketonic groups of DHAA and diketogulonic acid
(DKGA) has been the basis of many methods for the
determination of total vitamin C contents. Proteins
present in the samples are precipitated by adding
trichloroacetic acid (TCA) and aliquots of filtrate are
shaken with acid–washed charcoal (norit) or activated
charcoal55 to clarify the solutions and to oxidize AA to
DHAA. A reducing medium is produced by adding
thiourea prior to DNPH addition, otherwise unspecific
coloration is given by oxidants. The osazones
(λ max=545 nm) thus formed during the 3 h incubation at
891
37˚C by the reaction of DNPH and DHAA are dis-
solved by adding 85% H 2 SO 4 . Vitamin C can be
extracted with metaphosphoric acid–stannous chloride
solution without charcoal treatment for differential
determination of DKGA, DHAA and AA in the same
tissue extracts. The interference of sugars can be mini-
mized by carrying out incubation at 15˚C and measur-
ing the absorbance only after adding sulfuric acid for
75 min.56 The use of several acid mixtures has been
proposed for replacing the tedious dropwise addition of
sulfuric acid. Lack of specificity is found with many of
these methods; interfering osazones can be separated
by chromatographic methods such as TLC 57 and
HPLC58, but at the cost of making these procedures
tedious and cumbersome. The nature of DNPH meth-
ods for total vitamin C also makes it amenable to auto-
matic flow through analyses.59–61
Phenylhydrazinium chloride 62 produces a yellow
color (λ max=395 nm) when treated with ascorbic acid in
0.1 M HCl medium. The reaction contents are kept for
1 h in an incubator or water bath at 50±2˚C, thus mak-
ing the method time-consuming. Beer’s law is obeyed
in the range 25 – 100 µg of ascorbic acid. No interfer-
ence is observed from other vitamins, minerals,
glucose, sucrose, excipients and reducing agents.
However, the presence of excessive amounts of
riboflavin requires the addition of 0.5 g talc, which
imparts a yellow color to the solution. 3-Methyl-2-
benzothiazolone hydrazone63 reacts in the presence of
sodium metaperiodate to form a blue colored solution
(λ max=630 nm) which helps in the determination of
ascorbic acid over the range 6 – 14 meq ml–1.
Wang64 suggested the use of potassium iodate for the
determination of vitamin C in pharmaceuticals. The
absorbance is measured either in the UV region (288
nm) or in the visible region (445 nm). Besides aqueous
phase measurements, the yellow precipitate can be
extracted into chloroform65 (λ max=514 nm). The ICl2–
generated in the oxidation of AA by iodate 66 in acid
medium in the presence of Cl– ions has been used to
iodinate 2′,7′-dichlorofluorescein dye. The iodinated
dye ( λ max=525 nm) obeys Beer’s law up to 300 µg
(ε=8.81×103). Soft drinks67 have been analyzed using
the reaction of iodine in an acetic acid medium (λ max=
350 nm). Sirividya and Balasubramanian68 reported an
indirect procedure based on the oxidation of ascorbic
acid by a known excess of iodate in the presence of
acid for the analysis of pharmaceuticals and fresh fruit
juices. The unreacted iodate is used for hydroxylamine
oxidation to generate nitrite, which is then diazotized
with sulfanilic acid. The resulting diazonium salt is
coupled with N-(1-naphthyl)ethylenediamine dihy-
drochloride to form an azo dye (λ max=540 nm). The
procedure is a complicated one as it involves many
steps.
The reaction of hexacyanoferrate(III)69 (5) was used
for the determination of micro quantities of vitamin C
by measuring the decrease in color intensity of the
reagent (5) (λ max=420 nm) in McIlvaine buffer (pH 5.2)
892
solutions. Beer’s law is restricted within the range 180
– 270 µg of AA. A 200-fold amount of glucose, urea,
citric acid and tartaric acid; 50-fold excess of creatine
and 2-fold excess of creatinine do not interfere, but a
positive error is observed even with very small quanti-
ties of uric acid. In general, all such reagents that
reduce hexacyanoferrate(III) or oxidize hexacyanofer-
rate(II) under experimental conditions interfere.
Further the utility of the method is limited to colorless
solutions. Yet another method involving the oxidation
of phthalophenone to phenolphthalein by the reagent
(5) in alkaline solution was proposed by Al-Tamrah.70
This obeys Beer’s law up to 7 µg ml–1 (λ max=553 nm).
Sugars are tolerated only in microgram amounts. The
relative standard deviation and detection limit are
0.65% and 0.1 µg ml–1 respectively.
Direct UV spectrophotometry71–73 with background
correction methods such as thermal decomposition, UV
light irradiation, catalytic destruction and alkaline treat-
ment has been used for the determination of AA in soft
drinks, fruit juices and pharmaceuticals. However, the
rate of thermal decomposition is found to be very low72
and fruit juice samples that are unstable to alkaline
treatment, have fine particles, have a deep coloration or
contain high concentrations of caffeine, saccharin,
caramel and tannic acid can not be analyzed. Some
methods based on the Cu(II)-catalyzed oxidation are
reported for the assay of pharmaceuticals, fruits and
beverages74–77 allowing the determination of AA up to
120 µg ml–1 at λ max=267 nm. Fe(II) interferes seriously.
Only minute amounts of folic acid are tolerated. The
presence of Al(III), Mg(II) or Zn(II) gives a negative
error due to their catalytic effect.
Some methods involving the coinage metal (Cu, Ag,
Au) complexes have been worked out. The reduction
of Cu(II) in a biphasic system of isopentyl alcohol and
an aqueous solution of pH 4.6 to Cu(I), followed by its
complexation with cuproine to give a colored complex
(λ max=454 nm), was reported by Contreras et al.78 for
the analysis of foods and vegetables. Fresh fruits and
vegetables and dehydrated samples were analyzed after
extracting with 5% HPO3 and with a 1:1 mixture of
0.5% HPO3 and 0.05 M H2SO4 respectively. Also the
colored complexes of Cu(I) with 2,2-biquinoline 79
(λ max=540 nm), rhodanine80 (λ max=473 nm) and 2,9-
dimethyl-1,10-phenanthroline81–83 (λ max=450 nm) have
been used to determine ascorbic acid in different sam-
ples. However, the method using 2,9-dimethyl-1,10-
phenanthroline obeys Beer’s law over the range 2 – 20
µg ml–1, though it requires 1 h waiting time for full
color intensity. These methods based on the complex-
ation of reduced Cu(I) are rather unselective, since sub-
stances such as Fe(II), cysteine, or sodium thiosulfate
which lead to the reduction of Cu(II) to Cu(I) interfere
seriously. The gelatin complexes 84,85 of Ag(I)
(λ max=415 nm; ε=2.2×103) and Au(III) (λ max=540 nm;
ε=2.3×103) give colored products on adding AA to their
alkaline solutions. The procedure as suggested by Pal
et al.84 is not interfered with by glycine, alanine, fruc-
ANALYTICAL SCIENCES OCTOBER 1998, VOL. 14
tose, sucrose, citric acid, tartaric acid or other reducing
agents.
Analytical applications of Molybdenum Blue formed
on reduction of phosphomolybdate complex86, ammoni-
um molybdate87–89 or molybdic acid90 have been report-
ed by many workers for the determination of ascorbic
acid in pharmaceuticals, fruits and vegetables, pastries
and beverages. Ammonium molybdate–sulfuric acid
system requires 1 h for complete development of color
with ascorbic acid.87 However, such waiting time can
be decreased to 15 min by the addition of metaphos-
phoric acid–acetic acid solution.88 The colored species
obeys Beer’s law over the range 2 – 32 µg ml–1 at 760
nm (ε=4.3×104). Serious interferences are observed
due to phenolic compounds such as catechins, gallic
acid, pyrogallol and gallotannins; thiosulfate ions and
thiourea. Recently, P-Sb-Mo heteropoly acid91 has
been used to produce heteropoly blue (λ max=710 nm)
for the assay of ascorbic acid over the range 1 – 50 µg
ml–1 (ε=3.68×103). The use of folin reagent92 and folin
phenol93 (λ max=760 nm) has also been described for the
assay of biological samples after deproteinizing with
TCA. Beer’s law is obeyed up to 45 µg ml–1. The
color development is not obstructed by bovine serum
albumin, adenine, guanine, thymol and oxyhaemoglo-
bin. Folin-ciocalteu94 reagent reacts with ascorbic acid
to give a blue colored complex (λ max=730 nm) as well.
However, the method is time-consuming, as the full
color intensity requires 40 – 50 min. Ammonium meta-
vanadate95 gives a green color (λ max=680 nm) on heat-
ing for 10 min in the presence of ascorbic acid.
Though the method has been put to use for the analysis
of some samples, it is not sufficiently sensitive.
Many spectrophotometric methods based on the
reduction of Fe(III) to Fe(II) with ascorbic acid, fol-
lowed by the complexation of reduced Fe(II) with dif-
ferent reagents, have been reported. Amongst them,
α,α′-bipyridyl96–101 and 1,10-phenanthroline102–109 (o-
phen) find extensive use in the development of analyti-
cal procedures. Most of these methods are time-con-
suming, as full color development is achieved only
after waiting for 30 – 60 min. Micromodification97 of
the procedure applicable to human plasma and animal
tissue has been reported without the interference of glu-
cose, fructose, sucrose, glutathione and cysteine.
Recently, the procedure has been simplified by Arya
and Mahajan99 so as to require only 5 min waiting time,
instead of 30 or 60 min, with Beer’s law range up to 12
µg ml–1 (λ max=522 nm). Total ascorbic acid has been
determined in blood plasma100 after reducing DHAA
with dithiothreitol at pH 6.5 – 8.0, removing the excess
of dithiothreitol with N-ethylmaleimide and in urine101
by acidifying with TCA and shaking with activated
chorcoal. The reduced Fe(II) forms a water-soluble
colored complex with o-phenanthroline ( λ max=510 –
515 nm) at pH 1.5 – 6.5, with obedience of Beer’s law
up to 8 µg ml–1 (ε=2.2×104). Background correction104
as achieved by Cu(II)-catalyzed oxidation is necessary
for most samples, while the addition of NH4F106 as the
ANALYTICAL SCIENCES OCTOBER 1998, VOL. 14
inhibitor of light reduction of Fe(III)-phen complex is
needed in some cases. Selectivity for some of these
methods is poor. However, an improvement using
orange-red ferroin chelate in aqueous micellar medium
formed in the presence of the cationic surfactant
cetylpyridinium bromide 109 has been reported
(ε=2.6×104 at 510 nm). Ascorbic acid in fruits was
determined after extracting the ternary complex of
Fe(II) with α,α′-bipyridyl/o-phen and sulfophthalein110
dyes into chloroform (λ max=602 nm).
Many other compounds including oximes111–113 (6), 2-
oximinocyclohexanone thiosemicarbazone 114 (2-
OCHT) (7), 2-(5-bromo-2-pyridylazo)-5-dimethyl-
aminophenol115 (8) and 2-nitroso-5-(N-propyl-N-sulfo-
propylamino)phenol116 (9) have been investigated for
their use in the analysis of pharmaceuticals and biologi-
cal samples for ascorbic acid contents. The earlier
reported extraction111 of Fe(II)-dimethylglyoxime com-
plex into chloroform, which allows the determination
of 0.04 – 0.5 mM ascorbic acid, was modified by Arya
et al.112 They determined its concentration up to 14 µg
ml –1 at 514 nm. A proportionate decrease in color
intensity of Fe(III)-resacetophenone oxime113 complex
in sodium acetate–acetic acid buffer (pH 5) with the
increasing amounts of ascorbic acid was used for its
assay in the range 3.5 – 17.5 µg (ε=4×103). The method
using 2-OCHT determines ascorbic acid up to 12 µg
ml–1 (ε=1.49×104), but is interfered with by metal ions
such as Cu(II), Co(II), Ni(II) and Pd(II), in addition to
the interference caused by the oxalic acid, riboflavin,
oxidants and reductants. Color-forming reactions of
Fe(II) with ferrozine117–119 (λ max=562 nm) in acidic solu-
tions (pH 3 – 6), TPTZ 120–122 ( λ max =593, 595 nm),
quinaldic acid in presence of pyridine 123 ( λ max=380
nm), picolinic acid in presence of pyridine124 (λ max=400
nm) and nitroso-R salt125 ( λ max=705 nm) have been
used for the determination of vitamin C in a variety of
samples. The reagents picolinic acid and quinaldic
acid, when complexed with iron(II) in the presence of
pyridine, resulted in methods used successfully in the
analysis of pharmaceuticals, food products and biologi-
cal samples. The respective colored complexes getting
extracted into chloroform obey Beer’s law in the range
0.4 – 5.6 µg ml–1 and 2.5 – 25 µg ml–1 ascorbic acid
without the interference of common ingredients of the
samples studied. Though the method using ferrozine117
is not interfered with by sucrose, glucose, mannose,
fructose and formaldehyde, yet it suffers interferences
from tartaric acid, citric acid, Co(II), Ni(II) and Fe(II).
However, reactions of citric acid and tartaric acid can
be masked by adding Al(III) or La(III) ions and that of
iron(II) by passing the solution through a cation
exchanger.
Most of the reported methods based on the reducing
action of ascorbic acid on metal ions invariably make
use of an iron(III)–iron(II) redox system. A few use
copper(II)–copper(I), vanadium(V)–vanadium(IV) or
molybdenum/tungsten blue formation reactions, as
mentioned earlier in the text. Arya et al. have reported
893
a new redox system involving Cr(VI)-diphenyl-
carbazide complex 126 ( λ max=540 nm), which obeys
Beer’s law up to 3.2 µg ml–1. Common additives of
pharmaceutical preparations have no adverse effect on
the absorbance of the complex. Another fast and facile
method based on the proportionate decrease in
absorbance of iron(III)-ferronate complex127 (λ max=465
nm) by the addition of ascorbic acid was proposed by
the same authors after extracting the complex into
TBA/CHCl3 solution. Beer’s law is valid up to 10 µg
ml–1.
3 Conclusion
Even after the introduction of other instrument-based
procedures, photometric methods continue to be of
interest because of the ease in accessibility and their
quick applicability to the routine analyses. The molar
absorptivity for most of the colored species used in col-
orimetric analysis of vitamin C lies over the range 103
to 10 4 1 mol –1 cm–1 at the wavelength of maximum
absorbance. This enables the precise determination of
vitamin C in a variety of samples. The presence of cer-
tain substances, especially the matrix constituents, may
cause serious interferences. However, attempts to over-
come such interferences either by using masking agents
or making preliminary separations are invariably tried,
but sometimes without much success, thus resulting in
methods of varying selectivity. It has not been possible
to categorize the methods based on the selectivity since
the relevant data is found to be missing in the summary
part of most methods reported in Chemical Abstracts.
But none of the methods is found entirely specific for
vitamin C. Despite the reporting of several new photo-
metric methods, old procedures still continue to be
cited in different pharmacopoeias, indicating either the
lack of reliability or of general applicability of these
methods of vitamin C determination. Research workers
try to justify their work in terms of specific applica-
tions, but seldom give an comparative account with
other methods regarding analysis of particular type of
matrix. Therefore, to incorporate the comparative use
of such methods under specific analytical environment
requires some patience.
The authors wish to thank the Chairman, Department of
Chemistry, Kurukshetra University, Kurukshetra, for necessary
library facilities and Dr. Meenakshi Mahajan is grateful to CSIR
for financial assistance.
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(Received January 19, 1998)
(Accepted May 28, 1998)