World Journal of Pharmaceutical Research

Padarthi et al. SJIF Impact

World Journal of Pharmaceutical Factor 7.523
Research
Volume 6, Issue 5, 749-755. Research Article ISSN 2277–7105

SIMULTANEOUS ESTIMATION OF CAFFEINE AND BENZOIC ACID

IN SOFT DRINKS BY USING UV-VISIBLE SPECTROSCOPY

Vasubabu Padarthi*, Dr. K. N. Vidyadhar, Pudi Durgasyam, Yasarapu Manoj, Meka

Jhansy Saranya and Mokara Laxmi Priya

Department of Pharmaceutical Analysis, V. J’S College of Pharmacy, Rajahmundry.

Article Received on
25 Feb. 2017,
Revised on 17 March 2017,
Accepted on 06 April 2017
DOI: 10.20959/wjpr20175-8328
*Corresponding Author’
Vasubabu Padarthi
Department of
Pharmaceutical Analysis,
V. J’S College of
Pharmacy, Rajahmundry.
ABSTRACT
Five brands of soft drinks were analyzed for benzoic acid and caffeine
content by using simultaneous estimation method. The soft drink
includes Thums up, Limca, Coca cola, Sprite and Mountain dew. The
result shows that the pH of soft drinks ranges from 1.8 to 2.8. The
amount of benzoic acid and caffeine is determined by using
simultaneous estimation method. In this process, it was found that
amount of benzoic acid was low in Mountain dew. The caffeine
content in soft drinks was only found in Thums up, Coca cola and
Mountain dew and remaining other samples the presence of caffeine
absorbance under UV spectrophotometry are not detected.

KEYWORDS: Soft drinks, Benzoic acid, Caffeine, Simultaneous equation, UV

Spectrophotometry.

INTRODUCTION
A soft drink is a drink that typically contains carbonated water, a sweetener and a natural or
artificial flavoring. The sweetener may be sugar, high-fructose corn syrup, fruit juice, sugar
substitutes (in the case of diet drinks) or some combination of these. Soft drinks may also
contain caffeine, colorings, preservatives and other ingredients.

Benzoic acid and its salts are used as a preservative, represented by the E-numbers E210,
E211, E212 and E213. Benzoic acid inhibits the growth of mould, yeast and some bacteria. It
is used either directly or as its sodium, potassium or calcium salt.

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Padarthi et al. World Journal of Pharmaceutical Research

Caffeine which is chemically know as 1,3,7-Trimethylpurine-2,6-dione. Caffeine is a central

nervous system (CNS) stimulant, having the effect of temporarily warding off drowsiness and
restoring alertness.

It increases calcium permeability in sarcoplasmic reticulum, inhibiting phosphodiesterase

promoting accumulation of cyclic AMP. The most prominent is that it reversibly blocks the
action of adenosine on its receptor and consequently prevents the onset of drowsiness
induced by adenosine. Caffeine also stimulates certain portions of the autonomic nervous
system.

BENZOIC ACID CAFFEINE

MATERIALS AND METHODS

The materials used are Hydrochloric acid (Molychem), Distilled water, Caffeine (Spectrum
% purity: 98.5 %), Benzoic acid (Merck % purity: 99.9%). Which are brought from the local
store of Siri scientific, Rajahmundry.

Preparation of solutions
Preparation of stock solution of benzoic acid
Weigh 0.010 gm of benzoic acid and transferred with DI water in a 500ml volumetric flask. It
may be difficult to get the solid to dissolve, so you may want to start by putting the solid in a
500ml Erlenmeyer flask (not a volumetric flask), adding about 400ml of water, and heating
on a heat plate. You will then have to make a quantitative transfer of the entire solution to the
volumetric flask. Once the solid is dissolved in the volumetric flask, fill the flask to the mark
with DI water.

Preparation of stock solution of caffeine

Prepare a stock solution of caffeine by dissolving 0.022gm caffeine (accurately weighed and
transferred) with DI water in a 500 ml volumetric flask (you will probably want to use the

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 Padarthi et al. World Journal of Pharmaceutical Research

Sonicator to help dissolve the caffeine).Once the solid is dissolved fill the flask to the mark
with DI water.

Preparation of sample solutions: (Thums up, Limca, Sprite, Mountain dew, Coca cola)
Pour exactly 20 ml of sample into a 125 ml Erlenmeyer flask, then Sonicator the sample for 5
minutes to remove dissolved CO2 .Using the micro pipette (with a new tip) take 2 ml of
sample in 50 ml volumetric flask and make up the volume with 0.01M Hcl.

RESULTS AND DISCUSSIONS

THUMS UP

Fig no 1: Determination of caffeine and benzoic acid in Thums up.

λ for benzoic acid : 227
λ for caffeine : 272

Table no 1: λ max an absorbance values for caffeine and benzoic acid in Thums up.
BENZOIC A1%1cmBENZOIC
WAVELENGTH CAFFEINE THUMSUP A1%1cmCAFFEINE
ACID ACID
227 0.093 0.311 0.463 93 311
272 0.398 0.016 0.636 398 18

C1= a2λ2A1-a2λ1A2 b(a1λ1a2λ2-aλ1a1λ2)

C1=18 0.463-398 0.636 93 18-398 311
=8.334-253.128 1674-123778 = 0.002 C1= 20 mg
C2= a1λ1A2-a1λ2A1 b(a1λ1a2λ2-a2λ1a1λ2)

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Padarthi et al. World Journal of Pharmaceutical Research

=93 0.636-311 0.463 93 18-398

=840.845 122104 = 0.0069; C2=69mg

SPRITE

Fig no 2: Determination of caffeine and benzoic acid in sprite.

Table no 2: λ max and absorbance value for caffeine and benzoic acid in sprite.

BENZOIC A1%1cm A1%1cmBENZOIC

WAVELENGTH CAFFEINE SPRITE
ACID CAFFEINE ACID
227 0.093 0.311 0.421 93 311
272 0.398 0.018 0.039 398 18

COCO-COLA

Fig no 3: Determination caffeine and benzoic acid in coco-cola.

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Table no 3: λ max and absorbance values for caffeine and benzoic acid in coco cola.
BENZOIC A1%1cm A1%1cmBENZOIC
WAVELENGTH CAFFEINE COCACOLA
ACID CAFFEINE ACID
223 0.093 0.311 1.427 93 311
272 0.398 0.018 0.673 398 18

MOUNTAIN DEW

Fig no 4: Determination of benzoic acid and caffeine in mountain dew.

Table no 4: λ max and absorbance values for caffeine and benzoic acid in mountain
dew.
BENZOIC MOUNTAIN A1%1cm A1%1cm
WAVELENGTH CAFFEINE
ACID DEW CAFFEINE BENZOIC ACID
227 0.093 0.311 0.625 93 311
272 0.398 0.018 0.346 398 18

LIMCA

Fig no: 5 Determination of benzoic acid and caffeine in Limca

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Padarthi et al. World Journal of Pharmaceutical Research

Table no 5: λ max and absorbance values for caffeine and benzoic acid in Limca.
BENZOIC A1%1cm A1%1cmBENZOIC
WAVELENGTH CAFFEINE LIMCA
ACID CAFFEINE ACID
230 0.093 0.311 0.486 93 311
270 0.398 0.018 0.051 398 18

Benzoic acid and Caffeine

Table no 6: Amount of benzoic acid and caffeine in soft drinks.
Company Company
Benzoic acid Caffeine
standards standards for
Company concentration concentration
for benzoic caffeine (mg/l)
(mg/l) (mg/l) [24 25 26 27 28]
acid(mg/l)
Thumsup 20.0 69 70 52
Mountain dew 10.3 13.2 54 54
Limca 11.9 0.09 70 0
Coca-cola 19.8 31.2 46 46
Sprite 10.6 0.33 70 0

CONCLUSION
The analytical determination of benzoic acid and caffeine is not only important for quality
assurance purpose but also for consumer interest and protection. This work used is simple,
fast, and accurate UV-Visible spectroscopic method for the simultaneous determination of
benzoic acid and caffeine in five marketed samples of soft drinks Coca-cola, Thums up,
Sprite, Mountain dew, Limca. The results showed that the benzoic acid was found in all the
five soft drinks which are within the company standards. Whereas caffeine was found only in
Coca-cola, Thums up and Mountain dew which are also in company standards. So regular
checking of soft drinks should be done.

REFERENCES
1 H. Beckett, J. B. Stenlake. Simultaneous equations method: Practical pharmaceutical
chemistry. 4th edition part 2., New Delhi; CBS Publishers, 2007; 284-286.
2 Sandor Gorog. Multi component systems: Ultraviolet- visible spectrophotometry in
pharmaceutical analysis. Pharma Med press, 2011; 104-105.
3 Amos-Tautua, W. Bamidele Martin and E.R.E. Diepreye. Ultra-violet Spectrophotometric
Determination of Caffeine in Soft and Energy Drinks Available in Yenagoa, Nigeria.
Advance Journal of Food Science and Technology, 2014; 6(2): 155-158.
4 Doreen Amponsah. Determination of the amount of caffeine and benzoic acid in
selected soft drinks. International journal of scientific & engineering research, June 2014;
5: 1375.

www.wjpr.net Vol 6, Issue 5, 2017. 754

Padarthi et al. World Journal of Pharmaceutical Research

5 Zaifa Pan, Lili Wang, Weimin Mo, Wei Hu, Juan juan Zhang. Determination of
benzoic acid in soft drinks by gas chromatography with on-line pyrolytic methylation
technique. Analytica Chimica Acta, 2005; 545: 218–223.
6 M. A. Ford. The quantitative determination of benzoic acid in soft drinks by ion-exchange
chromatography. Analyst, 1966; 91: 15-22.
7 James K. Kusi, Samuel O. Levels of benzoic acid in soft drinks and fruit juices in Ghana.
IOSR Journal of Environmental Science, Toxicology and Food Technology (IOSR-
JESTFT) e-ISSN: 2319-2402, p- ISSN: 2319-2399, Dec 2014; 8: 36-39.
8 C.M. Lino , A. Pena .Occurrence of caffeine, saccharin, benzoic acid and sorbic acid in
soft drinks and nectars in Portugal and subsequent exposure assessment. Food Chemistry,
July 2010; 121: 503–508.
9 Qing-Chuan Chen, Jing Wang. Simultaneous determination of artificial sweeteners,
preservatives, caffeine, theobromine and theophylline in food and pharmaceutical
preparations by ion chromatography. Journal of Chromatography A, Dec 2001; 937: 1–2,
57–64.
10 Igelige Gerald1, David Ebuka Arthur1, Adebiyi Adedayo2. Paper Open Access
Determination of CaffeineIn Beverages. American Journal of Engineering Research
(AJER) e-ISSN : 2320-0847 p-ISSN ; 2320-0936, 2014; 3: 124-137.
11 Kathryn R. Williams, Alejandra Rodriguez Haralambides. Analysis of Soft Drinks: UV
Spectrophotometry, Liquid Chromatography and Capillary Electrophoresis. Journal of
chemical education, Feb 1966; 625: 15-22.
12 Pauline Maes, Yulia B. Monakhova, Thomas Kuballa, Helmut Reusch and Dirk W.
Lachenmeier. Qualitative and Quantitative Control of Carbonated Cola Beverages Using
1H NMR Spectroscopy. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY,
2012 Mar 21; 60(11): 2778-2784.
13 Nancy Karreman and Tom Zuzelski. Simultaneous Determination of Benzoic Acid and
Caffeine Concentrations by Single Wavelength. Isocratic High-Pressure Liquid
Chromatography. The Journal of experimental secondary sciences.
14 James k. kusi, Samuel o.Acquaah. Levels of benzoic acid in soft Drinks and Fruit Juices
in Ghana James. www.iosrjournals.org www.iosrjournals, Dec 2014; 8: 36-39.

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