A review on validation and determination of caffeine.docx

A REVIEW ON
DETERMINATION
AND VALIDATION
OF CAFFEINE
CONTENT IN
ENERGY DRINKS BY
RP-HPLC METHOD.
Sultan ul-uloom college of pharmacy

Department of Pharmaceutical Quality Assurance
Sultan ul-uloom college of pharmacy, Banjara Hills, Hyd. Page 2
A REVIEW ON DETERMINATION AND VALIDATION OF CAFFEINE
CONTENT IN ENERGY DRINKS BY RP-HPLC METHOD.
Dissertation submitted to the
Jawaharlal Nehru Technological University, Hyderabad.
In partial fulfillment of the requirements for the degree of
MASTER OF PHARMACY
By
RUQSAR FATIMA 19451S1411
Under the guidance of
Mrs. A. Madhavi, M. Pharm.
Assistant Professor
Department Of Pharmaceutical Quality Assurance
SULTAN UL-ULOOM COLLEGE OF PHARMACY,
(Recognized by AICTE & Pharmacy Council of India)
(Affiliated to J.N.T.U -Hyderabad)
Mount Pleasant, 8-2-249, Road No. 3, Banjara Hills, Hyderabad - 500 034,
Telangana State, India.

(0):040-23280222
Email: suucop@yahoo.com
Website: www.sultanuloompharmacy.ac.in
SULTAN-UL-ULOOM COLLEGE OF
PHARMACY
(AFFILIATED TO J.N.T.U-HYDERABAD)
(Estd. by Sultan Ul-Uloom Education Society)
Certificate
This is to certify that the dissertation work entitled “A REVIEW ON
DETERMINATION AND VALIDATION OF CAFFEINE
CONTENT IN ENERGY DRINKS BY RP-HPLC METHOD”
submitted to Jawaharlal Nehru Technological University, Hyderabad
in partial fulfillment for the award of the degree of Masters of Pharmacy
has been successfully carried out by RUQSAR FATIMA , H.T No.
19451S1411 during the academic year 2019-2020 under my personal
guidance and supervision in the Department of Quality Assurance, Sultan
Ul-Uloom College of Pharmacy, Mount Pleasant, Road No.3 , Banjara
Hills, Hyderabad.
This work is original and has not been submitted in any part or full for any
other degree of this or any other university.
I have a great pleasure in forwarding the same.
Signature of Supervisor Signature of External Examiner
Place:
Date:

(0):040-23280222
Email: suucop@yahoo.com
Website: www.sultanuloompharmacy.ac.in
SULTAN-UL-ULOOM COLLEGE OF
PHARMACY
(AFFILIATED TO J.N.T.U-HYDERABAD)
(Estd. by Sultan Ul-Uloom Education Society)
Certificate
This is to certify that the dissertation work entitled “DETERMINATION
AND VALIDATION OF CAFFEINE CONTENT IN ENERGY
DRINKS BY RP-HPLC METHOD” submitted to Jawaharlal Nehru
Technological University, Hyderabad in partial fulfillment for the award of the
degree of Masters of Pharmacy has been successfully carried out by RUQSAR
FATIMA , H.T No. 19451S1411 during the academic year 2019-2020 under the
personal guidance and supervision of Mrs. A. Madhavi, Assistant Professor in
the Department of Quality Assurance, Sultan-Ul-Uloom College of Pharmacy,
Mount Pleasant, Road No.3, Banjara Hills, Hyderabad.
This work is original and has not been submitted in any part or full for any
other degree of this or any other university.
I have a great pleasure in forwarding the same.
Place: PRINCIPAL
Date: Dr. ANUPAMA KONERU (M.PHARM., Ph.D.)

ABBREVIATIONS
RP-HPLC: Reversed-phase high-performance liquid chromatography.
UV: Ultraviolet.
FTIR: Fourier transform infrared.
LOD: Limit of detection.
LOQ: Limit of quantification.
mm: Millimetre.
µm: Micrometre.
v/v: Volume/volume.
nm: Nanometre.
r: Regression coefficient.

ABSTRACT
Coffee, Tea and soft drinks are very commonly used beverages in all over the world. The
caffeine is the main stimulant occurred in all drinks. Caffeine stimulates the central nervous
system, relaxation, myocardial stimulation, recreation etc. It can provide energy, decrease
fatigue, enhance performance etc. Caffeine having medicinal properties so can be used
along with other drugs for headache, stimulation, muscle relaxant etc. Up to certain limit
caffeine is useful but overdose of caffeine starts side effects on the human body. There are
various instrumental methods can be used for the determination of caffeine in plants, coffee,
tea, soft drinks and pharmaceutical formulation in presence of other drugs. HPLC
methods are the most common, reliable methods for the determination caffeine in complex
sample. Very low concentration of caffeine can be determined with high accuracy and
precision.
Here in this review we have summarized various HPLC methods used for the caffeine
analysis in various samples and complex mixtures with their chromatographic column,
mobile phase, flow rate, detector etc.
Keywords: Caffeine, HPLC analysis, Beverages, coffee.

CONTENTS
TITLE PAGE NO.
Introduction 9
Principle 11
Literature Review 12
Materials and Methods 16
Results and Discussion 21
Conclusion 25
Bibliography 26

LIST OF FIGURES AND TABLES
TITLE PAGE NO.
Structural form of Caffeine. 9
HPLC instrumentation. 11
Literature Review. 12
UV-Visible spectrum for standard
solution of Caffeine.
16
FT-IR spectra for standard Caffeine
(Sigma –Aldrich).
17
FT-IR spectra for standard Caffeine. 17
Chromatogram of standard solution of
Caffeine.
18
Marketed energy drink assay. 19-20
Linearity and regression characteristics
of standard Caffeine.
21
Calibration curve of standard solution
of Caffeine.
21
Precision for standard Caffeine. 22
Percentage recovery data for standard
drug.
23
Retention time of ten energy drinks
assayed and standard drug solution. 23-24
Limit of detection and Limit of
Quantification.
24

AIM & Objective: To give review on determination and validation of caffeine content in
energy drinks by using RP-HPLC method.
Introduction:
Naturally, Caffeine can be found in many leaves and seeds of many plants, there is about
more than 63 plant species around the world contain Caffeine. Its chemical name is 1, 3, 5-
trimethylxanthine and its chemical formula is C8H10N4O2. Caffeine in pure form can
appears as white, woolen mass, sparkling needles of powder. Caffeine molecular mass is
194.19 g/mole, and its melting point is 236°C. Sublimation degree is 178°C. Figure (1)
showed the structural form of Caffeine. The spread of the presence of caffeine in a variety of
plants played a major role in the long-term popularity of caffeinated products. The major
sources of caffeine are coffee, tea, guarana, cola nuts and cocoa. The measure of caffeine
found in these items differs, the most noteworthy sums are found in guarana (4-7%), tea
leaves (3.5%), coffee beans (1.1- 2.2%), cola nuts (1.5%) and cocoa beans (0.03%). A fatal
dose of caffeine has been calculated to be more than 10g (about 170mg kg−1 body weight).
The reported caffeine content in the main dietary sources varies significantly: 93.0–163.5mg
per cup in ground coffee, 46.7– 67.6mg per cup in instant coffee, 30.2 – 67.4mg per cup in
bag tea and 0.32–0.54mg/g in dark sweet chocolate. A previous study has provided that there
is no difficult health effect that can cause by acceptable level of individuals, namely (400–
450mg/day), (300mg/day), and (45mg/day) for healthy adults, women contemplating
pregnancy and young children age 4–6years, respectively. It was found that caffeine "1, 3, 7-
trimethyl-1H-purine-2, 6(3H, 7H)-Dione" has also been recognized to play a significant role
in terms of the pharmaceutical industries. It is pharmaceutically important due to its chemical
properties. For example, Caffeine is a weak base, cannot lose a proton form or act as acid at
pH value under 14. Furthermore, Caffeine acts as electrophile in position 1, 2 and 3. In blood,
there is no highly amount of protein bound in caffeine. Moreover, caffeine found in brain
with high concentration due to the lipophilic properties. The main techniques can use to
determine the levels of caffeine are LC-Spectrophotometric, Spectrophotometric method, and
HPLC.
Fig 1. Structural form of Caffeine.
INTRODUCTION

Physiological effects of caffeine to human:
Caffeine has numerous physiological effects on major organ systems, including the nervous
system, cardiovascular system, digestive system, and respiratory system. Renal function and
skeletal muscles are also affected by caffeine. Numerous studies have proven caffeine to be a
stimulant to human’s central nervous system (Spiller, 1998).It is also increase heartbeat rate,
dilate blood vessels and elevate levels of free fatty acids and glucose in plasma. 1 g of
caffeine leads to insomnia, nervousness, nausea, ear ringing, flashing of light delirium and
tremulousness. In cases of overdosing and in combination with alcohol, narcotics and some
other drugs, these compounds produce a toxic effect, sometimes with lethal outcome
(Mamina and Pershin, 2002; Ben Yuhas, 2002; Wanyikaet al., 2010; James et al., 1990;
Tavallali and Sheikhaei, 2009). Caffeine facilitates the conduction velocity in the heart and
directly affects the contractility of the heart and blood vessels. Nevertheless, caffeine may
significantly reduce cerebral blood flow by constricting of cerebral blood vessels. Caffeine
provides a diuretic effect due to elevating the blood flow and glomerular filtration rate of the
kidneys. Heartburn is an issue for some subjects’ gastrointestinal system after consuming
caffeine. The effects of caffeine to skeletal muscles are mainly the increasing occurrence of
tremors (James 1991; Spiller 1998).

Principle:
Fig 2. HPLC instrumentation.
To understand the principle of HPLC, we must first look at the principle behind the liquid
chromatography.
Liquid chromatography is a separation technique that involves:
 The placement (injection) of a small volume of liquid sample into a tube packed with
porous particles (stationary phase).
 Where individual components of the sample are transported along the packed tube
(column) by a liquid moved by gravity.
The main principle of separation is adsorption.
PRINCIPLE

S.NO AUTHOR
NAME
TITLE OF THE
ARTICLE
METHOD
PARAMETERS
VALIDATION
PARAMETERS
CONCLUSION
1. Raghad Saad
Hatam Bustan
et al.
Validation and
determination of
caffeine contents
in energy drinks
available on the
Iraqi market by
using HPLC
Column:(150mm x
4.6mm)
M.P:water:
methanol(40:60v/v)
Rt: 1.999 min
Flow rate:1 ml/min
Run time: 2.50 min
λ max : 273 nm
Linearity:1-200
µg/ml
Accuracy
(%Recovery):
98.091-100.500%
Precision(RSD%):
0.022-0.128%
LOD:0.579 µg/ml
LOQ:1.809 µg/ml
Robustness:
0.5 ml = 3.969
min
1.3 ml = 1.547
min
A reversed-phase
HPLC method
was developed
and validated
with UV
detection for the
determination of
Caffeine and
proved to be
more convenient
and effective for
the quality
control of
Caffeine in
energy drinks.
The method gave
good resolution
for Caffeine.
2. Ayat Abbood
& Dima
Aldiab et al.
HPLC
Determination of
caffeine in some
beverages and
pharmaceutical
dosage forms
available in
Syrian market
Column: ( 250mm
x 4.6 mm)
M.P:MeOH : water
(40:60 v/v)
Rt:3.9min
(paracetamol)
Rt:2.1min (aspirin)
Flow rate:1 ml/min
λ max: 272 nm
Linearity: 0.19-10
µg/ml
(r2
= 0.9985)
Accuracy: 97.15%
Precision: ≤ 10%
LOD:0.023 µg/ml
LOQ:0.065 µg/ml
The results show
that the HPLC
method
presented here
can be
considered
suitable for the
analytical
determination of
CAF in
beverages and
pharmaceutical
tablets, being
linear in the
concentration
range used, high
selectivity, high
precision and
adequate
accuracy at the
concentrations
studied.
LITERATURE REVIEW

3. Muchtaridi et
al.
Analytical method
development and
validation for the
determination of
caffeine in green
coffee beans
(Coffee Arabica
L.) from three
districts of west
Java, Indonesia by
HPLC
Column: (250 mm
x 4.6mm)
M.P: MeOH: water
(37:63)
Flowrate:1.0ml/min
Rt: 6.36 min
λ max: 274 nm
Linearity:1-200
ppm (r2
=0.998)
Accuracy:
90.723%-
102.853%
Precision: 0.192%
LOD: 9 ppm
LOQ: 28 ppm
The optimum
condition for the
analytical
method of
caffeine by
HPLC was
carried out. This
method meets
the acceptance
parameters of
validation.
4. Igelige Gerald
et al.
Determination of
caffeine in
beverages: A
review
*UV method
Apparatus: UV
spectrophotometer
(Jenway7305
spectrophotometer)
Standard solution:
1000 ppm (198.3
mg in 200 ml
water)
λ max: 276 nm
Concentration: 0.5-
70 ppm
Calibration
equation:
A= -0.032 + 1.01 x
104
C (mol1-1
)
r=0.999
n=9
*HPLC method
Column: C-18
(150mm x 4.6mm)
& Novapak C-18
(150mm x 3.9mm)
M.P: 20% : 80%
(v/v) acetonitrile
and deionised
water
λ max : 254 nm
Flow rate: 1ml/min
Run time: 4.1 min
Rt: 7.347 min
-
*HPLC method
Linearity:10 ppm-
100 ppm
(r2
=0.999)
Accuracy:99-
105%
Precision: 1.25%
LOD: 0.023 ppm
Variation
coefficient: 0.6%
The validated
HPLC method
for the
quantification of
caffeine in
beverages was
found to be
simple, precise,
accurate, and
allowed the
obtaining of
good results.
5. Mohammad
A.Abdulla et
al.
Determination of
caffeine, active
ingredient in
different coffee
drinks and its
characterization
*FTIR method
Apparatus: FTIR
Nicolet 6700
Conditions of
measurement:
Wavenumber: 400-
-
Caffeine content
in different
coffee drinks
was determined
by using FTIR
and TGA

by FTIR/ATR &
TGA/DTA
4000 cm-1
No: of scans: 32
Resolution : 4 cm-1
*TGA method
Temperature:
10ºC/min
% loss in mass at
200º C & 700º C is
noted.
-
method.TGA has
been included to
identify thermal
properties.
In FTIR method
the
wavenumbers
for different
coffee drinks
were noted;
Arabian coffee-
1643.44 cm-1
,
roasted coffee-
1648.69 cm-1
,
instant coffee-
1646.79 cm-1
.
In TGA method
% loss in mass at
200 ºC & 700 ºC
for instant coffee
is 8.63 & 68.62,
for roasted
coffee is 6.17 &
68.70, for
Arabian coffee is
5.05 & 78.09.
6. Ioannis
N.pasias et al.
Development of a
rapid method for
the determination
of caffeine in
coffee grains by
GC-FID- A fully
validated
approach
Column: Agilent
DB-1 (30mm
x0.32mm x1µm)
Injection volume:
1µl
Temperature:
280ºC
Flow rate: 1.8
ml/min
Detector
temp:300ºC
Linearity: 10-
1000 mg/L
(r2
=0.999)
Accuracy : ≥
93.5%
Precision : ≤ 5%
LOD: 3.1 mg/L
LOQ: 9.3 mg/L
Caffeine content
in coffee grains
was determined
by GC-FID
method. The
results obtained
were precise and
accurate.
7. Khalid A,
Ahmad S,
Raza H,
Batool M,
Lothi RK et al.
Determination of
caffeine in soft
and energy drinks
available in
market by using
UV-Vis
spectrophotometer
Apparatus: Perkin
Elmer Lambda 35
UV/Vis
spectrophotometer
Extracting solvent:
carbon
tetrachloride
λ max:270 nm
-
Caffeine was
determined in
soft and energy
drinks by
UV/Vis method.
The minimum
level of caffeine
in energy drinks
was found to be
32.04 mg/L and
the maximum

level was found
to be 101.705
mg/L. However
the minimum
level of caffeine
in soft drinks
was reported as
10.69
mg/serving and
the maximum
level was 42.17
mg/serving.

Materials and Chemical
The caffeine samples were collected from the different drug stores. In addition, ultra-high-
purity grade reagents were used for the dilution, preparation and analysis of samples so as to
avoid contamination at trace element levels. Double beam UV-Visible – Spectrophotometer -
1800, Shimadzu, (Japan). Equipped with quartz cell (1cm), High-performance Liquid
Chromatography (HPLC), UFLC -Shimadzu, CBM 20A, (Japan), Equipped with
HAISIL100A -C18 (150mm x 4.6mm, 5µm particle) analytical column, UV-Visible detector
and Column oven CTO-20A (4- 85)°C, Shimadzu (Japan), Digital Balance, Denver –TP-214,
(Germany), FT-IR, Bruker, TENSOR 27, (Germany), pH-meter, Hanna-pH211, (Romania)
and Ultrasonic cleaner, KQ200E, (Chain) were employed for the estimation.
Preparation of standard stock solution
Standard stock solutions of caffeine were prepared by weighing 0.01g from drug and the
solutions were then diluted to 50 ml with mobile phase using a polyethylene volumetric flask.
On each occasion the mixture was sonicated (15 minutes, 35 MHz). Standard stock solution
was further diluted with mobile phase to prepare serial solutions for standard curve.
Sample preparation
Different kinds of energy drinks were purchased from different Iraqi local supermarkets and
10 samples were analyzed using the indicated HPLC method. When test containers were
opened, the beverages were degassed and homogenized , then every sample was filtered by
using 0.45μm filter paper, filtered beverage tests of 2 mL were diluted to 20 times with
mobile phase.
Wavelength selection
Caffeine solutions at concentration of 10μg/mL in diluent were scanned by UV-Visible
spectrophotometer in the range of (200-400) nm. From the UV spectra, suitable wavelength
considered for monitoring the drug were 273nm on the basis of higher response.
Fig 3. UV-Visible spectrum for standard solution of Caffeine.
MATERIALS AND METHODS

FT-IR spectrum of Caffeine
FT-IR spectrum was recorded for Caffeine. The spectrum was compared with standard
spectrum in order to identify this compound.
Fig 4. FT-IR spectra for standard Caffeine (Sigma –Aldrich)
Fig 5. FT-IR spectra for standard Caffeine.
Chromatographic conditions of isocratic elution system
HPLC analysis was performed by isocratic elution. The flow rate was 1mL/min. The mobile
phase composition was water: methanol (40:60 v/v), adjusting pH to 5 by diluted (HCl /
NaOH). All injected solvent were filtered through 0.45µm filter paper and degassed by using
ultrasonic water bath , the volume of injected solvent was 20µL and the wavelength 273 nm.
Running time for overall analysis less than 3 minute as shown in figure (5).

Fig 6. Chromatogram of standard solution of Caffeine.
Calibration curve
Standard solutions containing Caffeine (1-200) µg/mL were prepared in the mobile phase. To
study the reproducibility of detector response at different level of concentration, 20µL of
standard solution was injected; calibration curve was obtained by plotting peak area against
concentration to calculate the calibration equation and correlation coefficients.
Optimization of HPLC method
Separation of caffeine was carried out by using by using a mixture of water- methanol with
the volume ratio (v/v) of (40:60), adjusting pH to 5 by diluted NaOH, at flow rate of
1mL/min with isocratic program and gave acceptable retention time of ( 1.999) min.
Validation of the method
Optimization of the HPLC instrument was performed daily and the instrument operating
parameters were made in order to achieve the maximum sensitivity as shown below:
Linearity and range
The linearity range was evaluated by using the standard solution prepared in the mobile
phase. The calibration range for caffeine was found to be 1 – 200 μg/mL. The calibration
graph was automatically drawn by plotting the value of peak area against the concentration of
caffeine.

Specificity
The specificity of the proposed method was evaluated by contrasting chromatogram acquired
from standards Caffeine and that from marketed solutions.
Limits of detection and Limit of quantitation
Sensitivity of the proposed method was assessed as far as (LOD) and (LOQ). "LOD = 3 SD/S
and LOQ = 10 SD /S, where S.D. is the standard deviation of y-intercept and S is the slope of
the line"
Effect of pH
The effect of pH variation of mobile phase on the selectivity and retention times was
determined by using mobile phase with pH from (3.00 to 6.00) under optimum condition.
Effect of variation of flow rate
Different flow rate were examined to determine its effect on separation of Caffeine under
optimum condition. HPLC system was injected with a standard solution prepared according
to the method of testing using flow rate (0.5, 0.7, 1.0 and 1.3) mL/min.
Analysis of a marketed energy drink
To determine the content of Caffeine in commercial energy drinks illustrated below in table
(1):
Table (1): Marketed energy drink assay.
Brand name Manufactured Labeled claim (μg/mL)
Red Bull AUSTRIA 320
Monster United States 338
Burn United States 320
Sting Nepal 330

Mountain Dew United States 152
Pepsi Max Ukraine, United
Kingdom, Italy,
Turkey, Australia.
129.1
Coca-Cola United States 96
Diet Coke United States 130
Pepsi United States 107
Thums Up India 90
HPLC system was injected with 20μL sample solution under chromatographic conditions of
the method. Peak areas were determined at wavelength 273nm and concentration of samples
were determined by using calibration curve obtained on the same HPLC system with same
condition by utilizing linear regression equation.

Validation of the method
Validation of the optimized HPLC method was carried out with respect to the following
parameter:
Calibration curve and linearity study
Caffeine showed good correlation coefficient in concentration range of (1-200) μg/ml. The
detector response over wide range of concentrations of analyte were plotted to obtain the
calibration curve figure (6). The square of the correlation coefficient and equation for the
curve is shown in table (2).
Table (2): Linearity and regression characteristics of standard Caffeine.
Parameters Linearity range
µg/mL
Regression equation Correlation
coefficient (r² )
Linearity range
µg/mL
1-200 Y = 33191x +
98839
r² = 0.9999
(r2
) value is greater than 0.9997. From this result, it is acceptable to use a single point
calibration in analysis of actual samples.
Fig 7. Calibration curve of standard solution of Caffeine.
RESULTS AND DISCUSSION

Precision
Precision can be determined by using the replicate analysis of caffeine sample. The values of
relative standard deviation (RSD %) was calculated and found not more than 1, and E% less
than 2 which indicates that the developed method is precise and reproducible [14].Results
were illustrated in table (3).
Table (3): Precision for standard Caffeine.
(µg/mL) R.S.D %
E%
Caffeine 20 0.022 1.930
50 0.128 1.411
150 0.022 0.472
Accuracy
Accuracy was examined by using three different concentrations for all samples. In this study,
the degree of agreement between a measured value and a true value was determined by using
the values of recoveries (%R, replicate analysis n = 3).

Table (4): Percentage recovery data for standard drug
From these acceptable limit of percentage recovery, Estimation of caffeine can be accurate
quantitatively.
Specificity
The retention time of the standard Caffeine was illustrated table (5).
Table (5): Retention time of ten energy drinks assayed and standard drug solution.
tR (min)
Standard Caffeine solution 1.999
Red Bull 2.009
Monster 2.021
Burn 2.006
Sting 2.030
Amount added
(µg/mL)
Amount found
(µg/mL)
Recovery [%]
Caffeine 20 19.620 98.091
50 50.710 101.501
150 150.500 100.500

Mountain Dew 2.011
Pepsi Max 2.000
Coca-Cola 2.026
Diet Coke 2.034
Pepsi 2.019
Thums Up 2.036
Result in table (5) showed that the retention time of the standard Caffeine and Caffeine in
energy drink solutions were very close, almost same, so the method was specific.
Limit of detection (LOD) and Limit of quantitation (LOQ)
The results of limit of detection and limit of quantification were illustrated in table (6). The
values indicate that the method is sensitive.
Table (6): Limit of detection and Limit of Quantification.
Regression
equation Slope
LOD
μg/mL
LOQ
μg/mL
Caffeine Y = 33191x +
98839
33191 0.597 1.809

This reversed-phase HPLC method was proved to be more convenient and effective for the
quality control of Caffeine in energy drinks. The method gave good resolution for Caffeine
with a short analysis time below 2.5 minutes. The technique used in these methods was
observed to be simple, conservative and helpful with acceptable LOD and LOQ. Rapidity and
ability of evaluating low concentration of Caffeine, made this method helpful for variety of
analyses, including pure medication investigation. The proposed methods did not use any
extraction process for recovering the Caffeine from the formulation excipients matrices
thereby, decreased the error, time for estimation of Caffeine and the general expense of the
investigation. The mobile phase used was simple with isocratic elution and low buffer
solution contrasted with the reported methods. The method is suitable for the determination
of Caffeine in energy drink without interference from commonly used excipients, and could
be used in a quality control laboratory for routine sample analysis.
CONCLUSION

1. Violeta N., Mira E." Original Research Paper Chromatographic Determination of Caffeine
Contents in soft and and Energy Drinks Available on The Romanian Market",1University of
Craiova, 2010,P.(351-353).
2. John M. Beale, John H. Block ,Wilson and Gisvold’s textbook of organic medicinal and
pharmaceutical chemistry,12th ed., Lippincott Williams and Wilkins, a Wolters Kluwer
business,china,2011.
3. Erdal Dinç , Filiz Yurtsever and Feyyaz Onur, Simultaneous determination of active
ingredients in binary mixtures containing Caffeine using liquid Chromatographic and
Spectrophotometric methods, Turkish J. Pharm. Sci.,2005;1(2):115-138.
4. Ayman M. Mohsen, Hayam M. Lotfy, Amr M. Badawey, Hesham Salem and Sonia Z.
Elkhateeb, application of three novel Spectrophotometric methods manipulating ratio spectra
for resolving a pharmaceutical mixture of Chlorpheniramine hydrochloride and caffeine,
IJPPS,2013; Vol-5:478- 487.
5. Kuldeep Delvadiya, Ritu Kimbahune, Prachi Kabra, Sunil K. and Pratik Patel,
Spectrophotometric Simultaneous analysis of Paracetamol, Propyphenazone and Caffeine in
tablet dosage forms, IJPPS,2011;Vol-3: 170-174.
6. Sonali S. Bharate and Sandip B. Bharate, Spectrophotometric and Chromatographic
determination of Acetylsalicylic acid and Caffeine in pure and in tablet dosage form, Journal
of Advanced Scientific Research, 2012; 3(1):73-81.
7. Sharma S., Sharma M. C., Sharma R., Sharma A. D., High Performance Liquid
Chromatographic Assay method for the determination of Paracetamol and Caffeine in Tablet
Formulation-in vitro dissolution studies, Journal of Pharmacy Research,2011;4(5):1559-1561.
8. Boyka G. T., Bistra D. K., Dimitar R. R., Lily P. P. and Ivanka P. P., HPLC Assay and
Stability Studies of Tablets containing Paracetamol and Caffeine, Int. J. Pharm. Sci. Rev.
Res.,2013;18(1):138-142.
9. Milkica C. Branka I., Sote V., Vesna K. and Zorica V., Statistical Optimization of Reverse
Phase High Performance Liquid Chromatography for the Analysis of Caffeine Paracetamol
and its Degradation Product paminophenol , Acta Chim. Slov., 2008; 55:665–670.
10. Violeta N., Mira E." Original Research Paper Chromatographic Determination of
Caffeine Contents in soft and and Energy Drinks Available on The Romanian Market",
1University of Craiova, 2010, P.(351-353).
11. Mei M., Mawahib E., Mohammed I.Badawi A. Abdalla A. ,"Determination of Caffeine in
Some Sudanese Beverages by High Performance Liquid Chromatography", University of
hartoum, 2012, P.(336-337 ).
BIBLIOGRAPHY

12. Uttam D. P., Abhijit V. N., Aruna V. S.,Tirumal A. D. and Kiran V. M., Simultaneous
Determination of Aceclofenac, Paracetamol and Chlorzoxazone by HPLC in Tablet Dose
Form, E-Journal of Chemistry, 2009 ,6(1), P.289-294.
13.Redasani V. K., Gorle A. P., Badhan R. A., Jain P. S. and Surana S. J., simultaneous
determination of Chlorpheniramine Maleate, Phenylephrine hydrochloride, Paracetamol and
Caffeine in pharmaceutical preparation by Rp-HPLC, R. C. Patel Institute of Pharmaceutical
Education and Research, 2012 ,P.1-22.
14.Hornedonunez A., Getek T. A., Korfmacher W. A. and Simenthal F., High-performance
liquid-chromatography of clindamycin and clindamycin phosphate with electrochemical
detection, Journal of Chromatography,1990, 503,P. 217-225.
15.Lioyd R. S., Joseph J. K. and John W. D., Introduction to modern liquid chromatography,
Third Edition , A John Wiley & Sons, Inc.,2010 .
16. Asensi D. R., Quesada S. MP., Valverde M. E., De Ramon G. E., Gomez M. J. and
Munoz C. I., Comparison of high-resolution liquid chromatography versus microparticle
enzyme immunoassay for the measurement of sirolimus levels in renal transplantation, Farm
Hosp, 2006, 30(3), P.142-8.
17. Braithwaith A. and Smith F. J., Chromatographic method, 5th edition,Kluwer academic
publisher, UK, 1999.

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