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: omprakashbhusnure@gmail.com

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

omprakashbhusnure@gmail.com

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.

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

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

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)