Talanta 51 (2000) 197 200

www.elsevier.com/locate/talanta

Short communication

The use of stabilizers in the UV assay of ascorbic acid

J.K. Kwakye *
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Uni6ersity of Science and Technology, Kumasi, Ghana
Received 5 February 1999; received in revised form 7 June 1999; accepted 3 August 1999

Abstract
Sodium thiosulphate (0.04%w/v) has been used to stabilize ascorbic acid in aqueous medium. The method has been
used to assay ascorbic acid in commercial tablet preparations. It is very accurate, precise and reproducible. It
compares favourably with official titrimetric method. The method is simple and can be adopted for the routine assay
of ascorbic acid in single component tablet formulations. 2000 Elsevier Science B.V. All rights reserved.
Keywords: Ascorbic acid; Stabilizer; Assay

1. Introduction
The use of ascorbic acid (vitamin C) cannot be
over emphasised. Therefore there is a the need to
find a quick and reliable method of assay.
Various methods have been employed for the
analysis of ascorbic acid in pharmaceutical formulations, fruit juices, urine, plasma etc. These include titrations [1,2], high performance liquid
chromatography [3], fluorimetry [4] etc. However,
the use of direct UV for the assay of ascorbic acid
in pharmaceutical formulations has not been easy
due to its instability in aqueous solutions. Recently a UV method of assay of ascorbic acid
based on its stability studies has been reported [5].
The method is very useful for ascorbic acid in the
* Tel.: +233-51-60351; fax: + 233-51-60359.
E-mail address: ustpharm@gh.com (J.K. Kwakye)

presence of other vitamins. However the method

is time dependent and needs strict carefulness to
achieve reliable and reproducible results.
The instability of ascorbic acid is due to its
oxidation to dehydroascorbic acid which is a reversible reaction and subsequently to 2,3-diketoL-gulonic acid. This later reaction is irreversible.
These reactions can be inhibited by antioxidants
(stabilizers). The effect of antioxidants is generally
to break up the chains formed during the propagation process by providing a hydrogen atom or
an electron [6]. Some of the commonly used antioxidants in aqueous systems are sodium sulphite, sodium metabisulphite, sodium bisulphite,
sodium thiosulphate, thiosorbitol and thiourea.
Based on availability, the potential of three of
these stabilizers viz. sodium sulphite (Na2SO3),
sodium metabisulphite (Na2S2O5) and sodium
thiosulphate (Na2S2O3), in stabilizing ascorbic

0039-9140/00/$ - see front matter 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 0 3 9 - 9 1 4 0 ( 9 9 ) 0 0 2 4 3 - X

198

J.K. Kwakye / Talanta 51 (2000) 197200

acid in aqueous medium has been explored. The

method was compared with titrimetric (official)
method.

2. Experimental

2.1. Materials
All absorbances were determined on CECIL
CE800 ultraviolet spectrophotometer using 1 cm
path length.
All formulated ascorbic acid tablets were purchased from the local market.
L-ascorbic acid from BDH chemicals, Poole,
England; sodium sulphite anhydrous AnalaR
BDH; sodium metabisulphite AnalaR BDH;
sodium thiosulphate anhydrous from Hopkin and
Williams, England; vitamin C tablets (100mg
Ascorbic acid) from; EFFAH Pharmaceutical,
Kumasi, Ghana, Batch No. 91R014, G Pharmaceuticals, Accra, Ghana, Batch No. VC44A,
Letap Pharmaceutical, Accra, Ghana, Batch No.
O15109, GIHOC Pharmaceutical, Accra, Ghana,
Batch No. 4040, Bikkai Laboratories, Kumasi,
Ghana, Batch No. 021577.

2.2. Methods
2.2.1. Effect of sodium sulphite and sodium
metabisulphite
Five concentrations of sodium sulphite in water
(0.01%, 0.02%, 0.04%, 0.08% and 0.16%w/v) or
sodium metabisulphite (0.02%, 0.04%, 0.1%,
0.2%, 0.4%w/v) in water were used to prepare
0.001%w/v of Ascorbic acid solutions. The absorbances at 266 nm were recorded at 5 min
intervals over a period of 60 min using the corresponding anti-oxidant blank solution.
2.2.2. Effect of sodium thiosulphate (Na2S2O3)
Three concentrations of sodium thiosulphate in
water (viz. 0.01, 0.02% and 0.04%w/v) were each
used to prepare 0.001%w/v Ascorbic acid solutions. The absorbances at 264 nm were recorded
at 5 min intervals over a period of 60 min for
each preparation using the corresponding sodium
thiosulphate blank solution.

2.2.3. Assay of ascorbic acid tablets

2.2.3.1. UV method using 0.04% sodium thiosulphate as sol6ent. An amount of powdered vitamin
C tablets equivalent to 0.01 g of ascorbic acid was
dissolved in 0.04%w/v Na2S2O3 in distilled water,
filtered and made up to 100 ml. 10 ml of the
resulting solution was further diluted to 100 ml
with the 0.04% sodium thiosulphate solution and
the absorbance at 264 nm determined using 0.04%
Sodium thiosulphate blank solution. The procedure was repeated using an accurately weighed
quantity (about 0.01 g) of pure L-ascorbic acid as
standard for comparison.
2.2.3.2. Titrimetric method. An amount of powdered tablets equivalent to 0.15 g of Ascorbic acid
was dissolved in a mixture of 30 ml of water and
20 ml of 1 M sulphuric acid. The mixture was
titrated with 0.1 M ammonium cerium (IV) sulphate using ferroin sulphate as indicator.

3. Results and discussion

The objective of this work was to determine a
suitable stabilizer and an optimum concentration
which would stabilize ascorbic acid long enough
for UV analysis to be carried out.
For sodium sulphite (see Table 1), the optimum
concentration required to achieve some degree of
stability was 0.02% w/v, but as the concentration
was increased beyond this value the rate of breakdown of the ascorbic acid increased. The inability
of the sodium sulphite to stabilize the ascorbic
acid within 1 h might be due to the high pH of
the medium. Ascorbic acid is indeed more stable
in acid medium than alkaline medium.
For sodium metabisulphite it was observed that
the degree of stability is better than that of
sodium sulphite, and as the concentration was
increased from 0.02% w/v to 0.4% w/v the rate of
decomposition of ascorbic acid was greatly reduced (Table 1).
For sodium thiosulphate changes in concentration of the stabilizer did not affect the magnitude

J.K. Kwakye / Talanta 51 (2000) 197200

of the absorbances. The stability was better than

any of the two stabilizers. At a concentration of
0.04% w/v sodium thiosulphate, there was virtually no change in absorbance over the one hour
period and hence rate of decomposition was zero.
This implies that the best of the three stabilizers
used was sodium thiosulphate at a concentration
of 0.04% w/v and this concentration was subsequently used in the assay of ascorbic acid in
commercial tablet preparations.
The assay of the commercial tablet preparations
gave results very much the same as those of the
official (Titrimetric) method (Table 2). There was
no significant difference between the two methods

199

with regard to accuracy and precision. The UV

method is however, simpler and faster.

4. Conclusions
Sodium thiosulphate at a concentration of
0.04% w/v is a suitable stabilizer for ascorbic acid
in a UV method of assay. The UV method of
assay using a stabilizer is very accurate, precise
and simple. It can be adopted for routine assay of
ascorbic acid tablets and raw materials where
local legislation does not enforce the use of an
official Pharmacopoeia method.

Table 1
Effect of different concentrations of stabilizers on ascorbic acid (0.001%w/v)
Concentration of stabilizers in
water (%w/v)

pH of medium (stabilizer/ascorbic acid

mixture at 25C

Rate of decomposition of ascorbic acid

(%/min104*

Sodium sulphite(Na2SO3)
0.01
0.02
0.04
0.08
0.16

7.4
8.8
8.1
8.2
8.7

1.913
1.889
2.538
1.910
3.493

Sodium metabisulphite (Na2S2O5)

0.02
0.04
0.10
0.20
0.40

4.3
4.8
4.5
4.2
4.0

2.494
2.385
1.511
0.849
0.581

Sodium Thiosulphate (Na2S2O3)

0.01
0.02
0.04

4.4
4.6
4.6

0.239
0.193
0.000

* Rate of decomposition of ascorbic acid was calculated from absorbance values using five samples of ascorbic acid (0.001%w/v)
in each case over a period of 60 min. The standard deviations ranged from 0.108 to 0.219. Each determination was repeated twice.
Table 2
Assay of ascorbic acid in commercial tablet preparations
Source of tablet preparation UV method

EFFAH
M&G
LETAP
GIHOC
BIKKAI

Titrimetric method

% Purity (n= 8)

Coefficient of variation (%)

% Purity (n =6)

Coefficient of variation (%)

105.4
102.8
100.7
101.2
103.0

0.30
0.29
0.28
0.30
0.29

105.5
102.7
100.8
101.3
103.1

0.11
0.11
0.10
0.11
0.11

200

J.K. Kwakye / Talanta 51 (2000) 197200

References
[1] British Pharmacopoeia, vol.1, British Pharmacopoeia
Commission, 1993, p. 54.
[2] British Pharmacopoeia, vol. II, British Pharmacopoeia
Commission, 1993, p.778.
[3] E.S. Wagner, B. Lindley, R.D. Coffin, High performance
liquid chromatographic determination of ascorbic acid in
urine., J. Chromatography 153 (1979) 225.

[4] M.J. Deutsch, C.E. Weeks, Micro fluorometric assay for

Vitamin C, J. Assoc. Off. Agric. Chem. 42 (1965) 248.
[5] J.K. Kwakye, Development of a UV spectrophotometric
method of assay of ascorbic acid (vitamin C) in pharmaceutical formulations, West Afr. J. Pharm. 11 (2) (1997)
65 75.
[6] L Lachman, H.A. Lieberman, J.L. Kanig. The Theory and
Practice of Industrial Pharmacy, second ed., Henry Kimpton, London, 1976.