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INTERNATIONAL JOURNAL OF PHARMACEUTICS & DRUG
VOL. 3 ISSUE 12, 2015; 399 – 402 ; http://ijpda.com; ISSN: 2348
RESEARCH ARTICLE
ESTIMATION OF CA
FEINE IN DIFFERENT
MARKETED BRANDS OF
SOFT DRINKS AVAILABLE
IN LATUR DISTRICT BY UV
SPECTROSCOPY TEC
NIQUE
Bhusnure O.G*1, Gholve S.B.1, Todkar V.B.,
R.R., Giram P.S.
1Channabasweshwar Pharmacy College, Dept of Quality
Assurance, Latur(MS), India 2Channabasweshwar Pharmacy College, Dept of Pharm
cutics, Latur(MS), India
Date Received: 4th December 2015; Date Accepted:
December 2015 Date published: 29th December
Email: [email protected]
Abstract: Five samples were chosen to include the co
monly consumed soft drinks like Coca Cola, Pepsi Cola,
Red Bull, and Thumps up, Sprite. The estimation is pe
formed on UV-1800 SHIMAZDU spectrophotometer co
trolled using the UV Probe version 2.42. It is shown from
the results of this study that the concentration of caffeine
in the beverage drinks obtained from the loca
Latur was significantly lower than the maximum auth
rized level. However since caffeine is an additive su
stance and because of health concerns arising from its
consumption, it seems appropriate that warning labels,
indication of the presence and amounts of caffeine should
accompany all caffeinated beverages.
Key words: Caffeine; Beverages; Soft drinks
tometry.
INTRODUCTION:
Caffeine is found in the leaves, seeds or fruits of over 63
plants species worldwide. The most common sources
caffeine are coffee, cocoa beans, cola nuts and tea leaves
and the worldwide consumption of products derived
from these natural materials means that caffeine is one of
PHARMACEUTICS & DRUG ANALYSIS
ISSN: 2348-8948
ESTIMATION OF CAF-
FEINE IN DIFFERENT
MARKETED BRANDS OF
SOFT DRINKS AVAILABLE
IN LATUR DISTRICT BY UV
SPECTROSCOPY TECH-
, Todkar V.B., Deshpande
Channabasweshwar Pharmacy College, Dept of Quality
Channabasweshwar Pharmacy College, Dept of Pharma-
2015; Date Accepted: 24th
December 2015
Five samples were chosen to include the com-
monly consumed soft drinks like Coca Cola, Pepsi Cola,
and Thumps up, Sprite. The estimation is per-
1800 SHIMAZDU spectrophotometer con-
trolled using the UV Probe version 2.42. It is shown from
the results of this study that the concentration of caffeine
in the beverage drinks obtained from the local market in
Latur was significantly lower than the maximum autho-
rized level. However since caffeine is an additive sub-
stance and because of health concerns arising from its
consumption, it seems appropriate that warning labels,
nd amounts of caffeine should
Soft drinks; Spectropho-
Caffeine is found in the leaves, seeds or fruits of over 63
plants species worldwide. The most common sources of
caffeine are coffee, cocoa beans, cola nuts and tea leaves
and the worldwide consumption of products derived
from these natural materials means that caffeine is one of
the most popular and commonly consumed drugs in the
world. Caffeine’s popularity ste
that it is a pharmacologically active substance and a mild
central nervous system stimulant.
that there is little risk of harm when a person consumes
less than 300mg of caffeine a day1, 2
Caffeine is a naturally occurring chemical stimulant called
trimethylxanthine. Its molecular formula is C8H10N4O2.
It is a drug and shares a number of traits with more not
rious drugs such as cocaine and heroin. Caffeine uses the
same biochemical mechanisms as those other dr
stimulate central nervous system (CNS)3. In its pure form,
caffeine is a white crystalline powder that tastes very bi
ter. It is moderately soluble in water at room temperature
(2 g/100 ml) but very soluble in boiling water (66
g/100ml)4. Caffeine does not contain any stereogenic ce
ters and hence is classified as an achiral molecule
Caffeine
Five samples were chosen to include the commonly co
sumed soft drinks like Coca Cola, Pepsi Cola, Red Bull,
and Thumps up, Sprite. The estimation is performed on
UV-1800 SHIMAZDU spectrophotometer controlled u
ing the UV Probe version 2.42. The software is easy to
handle and allows the user to emulate all measurement
tasks normally performed on the instrument with the a
ditional benefit of allowing data to be transferred to e
ternal folders.
MATERIAL AND METHOD
All the glassware were soaked overnight with chromic
acid solution and washed thoroughly with water and d
tergent, then rinsed with deionised water dried in the
oven and transferred to a desiccator before use. The
chemicals and reagents used in this study were of analyt
cal grade and no further purification was carried out and
was obtained Sd Fine Chemicals
Maharashtra, India. Pure anhydrous caffeine was o
tained from Coca Cola PLC, Nigeria. Different kinds of
commonly consumed soft and energy drinks including
Pepsi Cola, Diet Coca Cola, Mountain Dew, Coca Cola,
Bullet, Power Horse, Lucozade Boost and
purchased from the local market in Latur, India. The di
ferent brands of beverages were got from local market.
399
the most popular and commonly consumed drugs in the
world. Caffeine’s popularity stems mainly from the fact
that it is a pharmacologically active substance and a mild
central nervous system stimulant. It is generally agreed
that there is little risk of harm when a person consumes
less than 300mg of caffeine a day1, 2.
ally occurring chemical stimulant called
trimethylxanthine. Its molecular formula is C8H10N4O2.
It is a drug and shares a number of traits with more noto-
rious drugs such as cocaine and heroin. Caffeine uses the
same biochemical mechanisms as those other drugs to
stimulate central nervous system (CNS)3. In its pure form,
caffeine is a white crystalline powder that tastes very bit-
ter. It is moderately soluble in water at room temperature
(2 g/100 ml) but very soluble in boiling water (66
Caffeine does not contain any stereogenic cen-
ters and hence is classified as an achiral molecule
Caffeine
Five samples were chosen to include the commonly con-
oca Cola, Pepsi Cola, Red Bull,
The estimation is performed on
1800 SHIMAZDU spectrophotometer controlled us-
ing the UV Probe version 2.42. The software is easy to
handle and allows the user to emulate all measurement
ly performed on the instrument with the ad-
ditional benefit of allowing data to be transferred to ex-
All the glassware were soaked overnight with chromic
acid solution and washed thoroughly with water and de-
n rinsed with deionised water dried in the
oven and transferred to a desiccator before use. The
icals and reagents used in this study were of analyti-
cal grade and no further purification was carried out and
Sd Fine Chemicals Limited from Mumbai,
Maharashtra, India. Pure anhydrous caffeine was ob-
tained from Coca Cola PLC, Nigeria. Different kinds of
commonly consumed soft and energy drinks including
Pepsi Cola, Diet Coca Cola, Mountain Dew, Coca Cola,
Bullet, Power Horse, Lucozade Boost and Red Bull were
purchased from the local market in Latur, India. The dif-
ferent brands of beverages were got from local market.
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Bhusnure O.G et al,
The caffeine content of the different beverages was calc
lated by UV-1800 Spectrophotometer, Shimadzu.
A. Preparation of standard and sample solution
a. Standard stock solution preparation
The standard stock solution of caffeine was prepared by
weighing 0.02g of caffeine and was dissolved in 1000 mL
of distilled water and got 10 mL of this solution and d
luted up to 100 mL with distilled water.
After preparing the stock solution the dilutions were pr
pared as:
1 mL of stock solution dissolved in 40 mL of distilled w
ter in first volumetric flask and 5 μg/mL dilution was
prepared. 1 ml of stock solution was taken and dissolved
in 20 mL of distilled water in second volumetric flask and
10 μg/mL dilution was prepared. 1 mL of stock solution
was dissolved in 13.33 mL of distilled water in third v
lumetric flask and 15 μg/mL dilution was prepared. 1 mL
of stock solution was dissolved in 10 mL of distilled water
in fourth volumetric flask and 20 μg/mL dilution was
prepared. 1 ml of stock solution was dissolved in 8 mL of
distilled water in fifth volumetric flask and 25μg/mL dil
tion was prepared.
1 mL of stock solution was dissolved in 5.7 mL of distilled
water in seventh volumetric flask and 35 μg/mL dilution
was prepared. 1 ml of stock solution was dissolved in 5
mL of distilled water in eighth volumetric flask and 40
μg/mL dilution was prepared.
b. Sample solution preparation
1 mL of beverage sample was diluted up to 100 mL with
distilled water and took the absorbance at 272 nm. The
absorbencies of these different solutions were shown in
Table 2.
B. Determination of Wavelength
The was determined by scanning the standard solut
from 190-400 nm range. We got the maximum absorbance
272 nm. This value was used for calculating the absorbe
cies of different dilutions. The absorbencies of different
dilutions are given in Table 1. Using the data obtained
from UV spectrophotometer for the different dilutions of
standard stock solution, a calibration curve got by plo
ting absorbance versus concentration of dilutions. The
calibration curve is shown in fig1.
Graphs and Table:
The standard linear calibration curve obtained from the
standard solutions analysis is presented in Fig. 2. It
showed a good linear relationship between the abso
bance and concentrations of the standard solutions. The
caffeine content levels in beverage samples are presented
Bhusnure O.G et al, Int J. Pharm. Drug. Anal, Vol: 3, Issue: 12, 2015; 399-402
Available online at http://ijpda.com
The caffeine content of the different beverages was calcu-
Spectrophotometer, Shimadzu.
and sample solution
The standard stock solution of caffeine was prepared by
weighing 0.02g of caffeine and was dissolved in 1000 mL
of distilled water and got 10 mL of this solution and di-
After preparing the stock solution the dilutions were pre-
1 mL of stock solution dissolved in 40 mL of distilled wa-
ter in first volumetric flask and 5 μg/mL dilution was
prepared. 1 ml of stock solution was taken and dissolved
n 20 mL of distilled water in second volumetric flask and
10 μg/mL dilution was prepared. 1 mL of stock solution
was dissolved in 13.33 mL of distilled water in third vo-
lumetric flask and 15 μg/mL dilution was prepared. 1 mL
in 10 mL of distilled water
in fourth volumetric flask and 20 μg/mL dilution was
prepared. 1 ml of stock solution was dissolved in 8 mL of
distilled water in fifth volumetric flask and 25μg/mL dilu-
in 5.7 mL of distilled
water in seventh volumetric flask and 35 μg/mL dilution
was prepared. 1 ml of stock solution was dissolved in 5
mL of distilled water in eighth volumetric flask and 40
mL of beverage sample was diluted up to 100 mL with
distilled water and took the absorbance at 272 nm. The
absorbencies of these different solutions were shown in
The was determined by scanning the standard solution
400 nm range. We got the maximum absorbance
272 nm. This value was used for calculating the absorben-
cies of different dilutions. The absorbencies of different
dilutions are given in Table 1. Using the data obtained
r the different dilutions of
standard stock solution, a calibration curve got by plot-
ting absorbance versus concentration of dilutions. The
The standard linear calibration curve obtained from the
standard solutions analysis is presented in Fig. 2. It
showed a good linear relationship between the absor-
bance and concentrations of the standard solutions. The
caffeine content levels in beverage samples are presented
and illustrated in Table 1 and Fig.
Sr. No.
Concentration
μg/ml
1 15
2 20
3 30
4 40
5 50
Table 1: Absorbencies of dilutions
Fig1: Calibration curve between Absorbance and Co
centrations
Estimation of Caffeine by UV Spectroscopic Method
1) Caffeine 15ug/ml:
2)Caffeine 20 ug/ml:
200.00 250.00 300.00
Ab
s.
0.667
0.600
0.400
0.200
0.000
-0.069
nm.
200.00 250.00 300.00
Ab
s.
0.928
0.500
0.000
-0.095
400
and illustrated in Table 1 and Fig. 3.
Absorbance
0.223
0.315
0.468
0.661
0.826
Table 1: Absorbencies of dilutions
Calibration curve between Absorbance and Con-
centrations
Estimation of Caffeine by UV Spectroscopic Method
nm.
300.00 350.00 400.00
nm.
300.00 350.00 400.00
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Bhusnure O.G et al, Int J. Pharm. Drug. Anal, Vol: 3, Issue: 12, 2015; 399-402
Available online at http://ijpda.com
401
3)Caffeine 30 ug/ml:
4)Caffeine 40 ug/ml:
5)Caffeine 50ug/ml:
Overlay graph of standard Caffeine:
Beverages/Soft drinks/ Energy drinks:
Brand Name Absorbance Caffeine content per
serving ( mg)
Coca-cola 0.153 6.77
Pepsi 0.125 5.53
Red bull 0.197 8.72
Thumps up 0.246 10.89
Sprite 0.002 0.088
Monster 0.329 14.05
Sting 0.234 9.29
Table 2: Caffeine content in different
Estimation of Caffeine By UV Spectroscopic Method
1) Pepsi:
2) Coca cola:
3) Red bull:
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
1.400
1.000
0.500
0.000
-0.139
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
2.011
1.500
1.000
0.500
0.000
-0.192
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
2.338
2.000
1.000
0.000
-0.224
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
5.000
4.000
3.000
2.000
1.000
0.000
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
0.743
0.500
0.000
-0.422
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
0.890
0.500
0.000
-0.401
nm.
200.00 250.00 300.00 350.00 400.00
Ab
s.
0.212
0.000
-0.200
-0.400
-0.470
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Bhusnure O.G et al,
4) Thumps up:
Fig. 3: Chart showing caffeine contents in the beverage samples
CONCLUSION
It is shown from the results of this study that the conce
tration of caffeine in the beverage drinks obtained from
the local market in Latur was significantly lower than the
maximum authorized level. However since caffeine is an
additive substance and because of health concerns arising
from its consumption, it seems appropriate that wa
labels, indication of the presence and amounts of caffeine
should accompany all caffeinated beverages. None of the
drinks evaluated in this study were so labeled. The
UV/VIS spectrophotometric method employed in this
study for the quantification of caffeine in beverages was
found to be relatively easy, fast and cheap. The major i
strument required is a modern computerized UV/VIS
spectrophotometer which can be acquired at an affordable
price. This analytical method may therefore, be reco
mended for the rapid, accurate and sensitive quantific
tion of caffeine in beverages by any educational instit
tions in developing countries.
ACKNOWLEDGEMENT
Authors are thankful to the Principal, Dr. Thonte S.S.,
Channabasweshwar Pharmacy College, Latur (MS), India
nm.
200.00 250.00 300.00 350.00
Ab
s.
0.532
0.000
-1.000
-2.000
-2.410
Caffeine content per serving (
mg)
Ca
ffe
ine
co
nte
nt(
)mg
/Se
rve
Bhusnure O.G et al, Int J. Pharm. Drug. Anal, Vol: 3, Issue: 12, 2015; 399-402
Available online at http://ijpda.com
5) Sprite:
Fig. 3: Chart showing caffeine contents in the beverage samples
It is shown from the results of this study that the concen-
the beverage drinks obtained from
the local market in Latur was significantly lower than the
maximum authorized level. However since caffeine is an
additive substance and because of health concerns arising
from its consumption, it seems appropriate that warning
indication of the presence and amounts of caffeine
should accompany all caffeinated beverages. None of the
drinks evaluated in this study were so labeled. The
UV/VIS spectrophotometric method employed in this
caffeine in beverages was
found to be relatively easy, fast and cheap. The major in-
strument required is a modern computerized UV/VIS
spectrophotometer which can be acquired at an affordable
price. This analytical method may therefore, be recom-
e rapid, accurate and sensitive quantifica-
tion of caffeine in beverages by any educational institu-
Authors are thankful to the Principal, Dr. Thonte S.S.,
Channabasweshwar Pharmacy College, Latur (MS), India
for his constant inspiration and support for all facility.
REFERENCES
1. Smith AP., "Caffeine at Work." Hum Psych
pharmacol. 2005 Aug;20(6):441.
2. Rogers PJ, Dernoncourt C. "Regular caffeine co
sumption: a balance of adverse and beneficial e
fects for mood and psychomotor performance."
Pharmacol Biochem Behav. 1998 Apr;59(4):1039
45.
3. S. Bolton, G. Null, Caffeine, psychological effects,
use and abuse. Orthomolecular Psychiatry; 1981,
10(3): 202-211.
4. Susan Budavari, The Merck Index, Whitehouse
Station, NJ: Merck & Co., Inc. 1996, Ed. 12th p.
1674.
5. L. Klosterman, The Facts about caffeine (Drugs).
Benchmark Books (NY), 2006, p. 43. ISBN 0
2242-0.
6. T. Vallombroso, Organic Chemistry Pearls of
Wisdom. Boston Medical Publishing Corp., 2001,
p. 43. ISBN 1-58409-016
400.00 200.00 250.00 300.00
Ab
s.
0.274
0.000
-1.000
-2.000
-2.916
0
2
4
6
8
10
12
14
16
Coca-
cola
Pepsi Red
bull
Thum
ps up
Sprite Mons
ter
Caffeine content per serving ( 6.77 5.53 8.72 10.89 0.088 14.05
Caffeine content per serving (mg)
402
r his constant inspiration and support for all facility.
Smith AP., "Caffeine at Work." Hum Psycho-
pharmacol. 2005 Aug;20(6):441.
Rogers PJ, Dernoncourt C. "Regular caffeine con-
sumption: a balance of adverse and beneficial ef-
sychomotor performance."
Pharmacol Biochem Behav. 1998 Apr;59(4):1039-
S. Bolton, G. Null, Caffeine, psychological effects,
use and abuse. Orthomolecular Psychiatry; 1981,
Susan Budavari, The Merck Index, Whitehouse
& Co., Inc. 1996, Ed. 12th p.
L. Klosterman, The Facts about caffeine (Drugs).
Benchmark Books (NY), 2006, p. 43. ISBN 0-7614-
T. Vallombroso, Organic Chemistry Pearls of
Wisdom. Boston Medical Publishing Corp., 2001,
6-2.
nm.
300.00 350.00 400.00
Sting
9.29
Caffeine content per serving (mg)