DETERMINATION OF ASCORBIC ACID CONTENT BY

IODOMETRY BY USING STANDARD SOLUTION AND

DETERMINATION OF CAFFEIN CONTENT

By :
Name : edi wardana naibaho (4173131007)
sri rahmadani (417)
luxy grebers swend sinaga (417)
class : chemistry education 2017
courese : experiment of quantitative and qualitative analitycal
chemistry

CHEMISTRY EDUCATION
CHEMIISTRY
FACULTY OF MATHEMATIC AND SCIENCE
UNIVERSITY OF MEDAN
MEDAN
2019
A. Title : Determination Of Ascorbic Acid Content By Iodometry By
Using Standard Solution And Determination Of Caffein Content
B. Objectives : To determine the level of ascorbic acid using iodimetry
titration using iodine standard solution as its titrant and determine the level of
a caffeine by using iodometric titration using a standard solution of Na2S2O3 as
its titrant.
C. Theoretical review:

Vitamin C or ascorbic acid is essential for human life and is required for a
range of physiological functions in human body. It can be found either in fresh fruits
and vegetables naturally or in medical forms such as normal tablets, effervescent
tablets and liquid vials. It is the most widely taken supplement. Though daily
requirements of vitamin C are changeable according to the age, sex and conditions,
it is around 75 to 90 mg per day for healthy adults and no more than 2000mg per
day is recommended1 . It is one of the most ubiquitous vitamins ever discovered.
Besides plays a paramount role as an antioxidant and free radical scavenger, it has
been suggested to be an effective antiviral agent (Chowdhury,2016).

In addition, ascorbic acid has been widely used in the pharmaceutical,

chemical, cosmetic and food industry as antioxidant. Therefore, there is a need to
find an accurate, reliable, rapid, and easy-to implement method for measuring the
amount of ascorbic acid in a sample. However, there have been difficulties in
quantifying ascorbic acid due to its instability in aqueous solution. The instability
of ascorbic acid is due to its oxidation to dehydroascorbic acid, which is a reversible
reaction, and subsequently to 2,3-diketo-L-gulonic acid. The later reaction is
irreversible (Chowdhury,2016).

(Chowdhury,2016).
 The chemical methods for the determination of ascorbic acid (vitamin C)
have been mainly' based on the method of Tillmans, Hirsch, and Hirsch employing
2,6-dichlorophenolindophenol or on some modification of the iodometric titration.
In general the iodometric methods have been subject to criticism since they lack
specificity and a sharp end point, although Stevens has shown that by employing a
double back-titration in the presence of a high concentration of hydrogen ions the
iodine end point may be made very sharp. Further, Tauber and Kleiner have
demonstrated that the iodometric method is adequate for the determination of
ascorbic acid in citrus fruit juices, since interfering substances are absent. However,
in common with all iodometric methods, the reagents employed in Stevens’ method
change their titer with time, requiring frequent standardization, and the double
back-titration increases the volumetric error (Ballentine,1941).

The main advantages of the iodometric titration method are its simplicity,
the use of very elementary equipment, easily available reagents of low cost and
speed of reaction of iodine with L-AA. In the present study, since the L-AA
amounts of all analyzed fruit/vegetables were reasonably high, LOD and LOQ are
not essential issues. However, in some highly colored extracts it is difficult to
accurately determine the endpoint of titration. The LOD and the LOQ of L-AA
content were 0.9 and 2.9 mg/mL, respectively. Suntornsuk et al. (2002) validated
and applied a similar iodometric titration method to herbal juices, finding higher
value limits. Overall, both methods showed much more restrictive limits than those
we determined for UHPLC-PDA (22 and 67 ng/mL for LOD and LOQ,
respectively) (Spínola et al., 2012). Besides that, iodometric titration presents the
inconvenience of exposing samples to light and oxygen during titration which can
lead to L-AA degradation and the method is susceptible to co-extracted
interferences and may overestimate L-AA due to the presence of oxidizable species
other than L-AA. Moreover, initial DHAA is never quantified in this method since
L-AA is oxidized to DHAA by iodine (Spínola,2013).

LC revealed to be a more specific, selective and sensitive technique for

determination of L-AA in the different foodstuffs. Moreover, this method requires
less reagents and material, is less time consuming than the titration method, less
susceptible to systematic errors and allows the quantification of total vitamin C
content. Since the UHPLC-PDA methodology requires a large financial investment
in equipment, the iodometric titration provided satisfactory quantitative results
(according to the correlation measurements) and can be applied in a preliminary
analysis or in situations where equipment cost is an obstacle but availability of
human resources is not (Spínola,2013).

Caffeine an alkaloid of the methylxanthine family is a naturally occurring

substance found in the leaves, seeds or fruits of over 63 plants species worldwide.
The most commonly known sources of caffeine are coffee, cocoa beans, cola nuts
and tea leaves. In its pure state, it is an intensely bitter white powder. Its chemical
formula is C8H10N4O2, its systematic name is 1, 3, 5- trimethylxanthine (). Its
structural formula is as shown below.

Caffeine is a pharmacologically active substance and depending on the dose,

can be a mild central nervous system stimulant. Caffeine does not accumulate in
the body over the course of time and is normally excreted within several hours of
consumption (Wanyika,2010).

Other naturally occurring methylxanthines include theobromine and

theophyline. Methylation of theobromine forms caffeine. Caffeine and related
compounds contain an imidazole ring fused to a pyrimidine ring (Wanyika,2010).

Many analytical methods have been developed for the determination of

cafeine in cofee beans and products containing cafeine including electroanalytical
; chromatographic techniques including gas chromatography, high performance
liquid chromatography (HPLC), liquid chromatography-particle beam/electron
ionization mass spectrophotometry, liquid chromatography-tandem mass
spectrometry, and spectroscopic techniques including nuclear magnetic resonance
spectroscopy, near infrared spectroscopy, near infra-red refectance spectroscopy,
and UV–Vis spectroscopy, and fuorescence polarization immunoassays. HPLC is
the method of choice by many researchers in determining the cafeine contents of
beverages, tea leaves and cofee beans. As diferent literatures indicated
spectrophotometric determination of cafeine is also reported as preferred method of
determination such as UV–Vis spectrophotometry because of its relatively low cost,
rapidity, high. accuracy and reproducibility. But UV–Vis spectrophotometric
method cannot be used directly for determination of cafeine in cofee beans extracted
with water owing to the matrix efect of UV–Vis absorbing substances in the sample
matrix (Weldegebrea,2017).

In aqueous solution of cofee beans it was observed that there is spectral

interference from caffeine and chlorogenic acid in the wavelength regions of 200–
500 nm. Yet this method requires the extraction of cafeine from the aqueous
solution of cofee beans using dichloromethane for the spectroscopic determination.
Tis is necessary since the cafeine spectrum is overlapped with other compounds
found in cofee. Hence, the use of dichloromethane limits the wider application of
UV–Vis method (Weldegebrea,2017).

D. Methode : Iodometry Titration

E. References :

Ballentine, R. (1941). Determination of Ascorbic Acid in Citrus Fruit Juices. Ind.

Eng. Chem. Anal. , 13 (2), pp 89–89.
Blen Weldegebreal, M. R.-A. (2017). Development of new analytical methods for
the determination of cafeine content in aqueous solution of green cofee
beans. Chemistry Central Journal, 11:126.
Chowdhury, M. s. (2016, march 1). research gate. Retrieved from researchgate:
www.researchgate.net
Spínola, V. (2013). Effect of time and temperature on vitamin C stability in
horticultural extracts UHPLC-PDA vs iodometric titration as analytical
methods. Food Science and Technology , 489-495.
Wanyika, H. N. (2010). Determination of caffeine content of tea and instant
coffee brands found in the Kenyan market. African Journal of Food
Science , Vol. 4(6), pp. 353 – 358.