A NOVEL RP-HPLC METHOD FOR THE QUANTIFICATION OF ESOMEPRAZOLE IN FORMULATIONS

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AVN. Gupta et al., IJSID 2011, 1 (2), 165-171

ISSN:2249-5347
IJSID
International Journal of Science Innovations and Discoveries An International peer
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Research Article Available online through www.ijsidonline.info
A NOVEL RP-HPLC METHOD FOR THE QUANTIFICATION OF ESOMEPRAZOLE IN
FORMULATIONS

A.V.N Guptha, A. Babu, D. Masatan, G. Srilatha, P. Sambaiah, CH. Koteswara Rao, P. Parvathi*
Dept of P.G Chemistry, S.S.N College, Narasaraopeta, Andrapradesh, India

Received: 03.08.2011

Modified: 14.10.2011

Published: 27.10.2011
ABSTRACT
A simple accurate RP-HPLC method was developed and
validated for rapid assay of Esomeprazole in formulation. Isocratic
*Corresponding Author
pump elution at a flow rate of 0.8ml/min was employed on
symmetry Chromosil C18, 250x4.6mm, 5µm specification at 27. c
temperature. The mobile phase consisted of Methanol: ACN: THF
60:30:10 (V/V). The UV detection wavelength was 272 nm and
20µl sample was injected. The retention time for Esomeprazole
was 6.05 min. The percentage RSD for precision and accuracy of
Address: the method was found to be less than 2%. The method was

Name: P. Parvathi validated as per the ICH guidelines.
Key Words: Esomeprazole, RP-HPLC, UV detection, recovery,
Place: Guntur, AP, India
precise, 272 nm
Email:

parvathi.pothuri@gmail.com

INTRODUCTION

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INTRODUCTION
Esomeprazole is a proton pump inhibitor which is used in the treatment of dyspepsia, peptic ulcer
disease (PUD), gastroesophageal reflux disease (GORD/GERD) and Zollinger-Ellison syndrome.
Esomeprazole reduces acid secretion through inhibition of ATPase in gastric parietal cells. By inhibiting
the functioning of this enzyme, the drug prevents formation of gastric acid. Common side effects include
headache, diarrhoea, nausea, gas, decreased appetite, constipation, dry mouth, and abdominal pain. More
severe side effects are severe allergic reactions, chest pain, dark urine, fast heartbeat, fever, paresthesia,
persistent sore throat, severe stomach pain, unusual bruising or bleeding, unusual tiredness, and
yellowing of the eyes or skin. Esomeprazole is a competitive inhibitor of the enzymes CYP2C19 and
CYP2C9, and may therefore interact with drugs that depend on them for metabolism, such as diazepam
and warfarin. The drug is rapidly cleared from the body, largely by urinary excretion of
pharmacologically-inactive metabolites such as 5-hydroxymethylesomeprazole and 5-
carboxyesomeprazole[4]

Figure.1: Chemical structure of Esomeprazole
EXPERIMENTAL
Materials
Working standard of Esomeprazole was obtained from well reputed research laboratories. HPLC
grade water, methanol, Acetonitrile ,Tetrahydrofuran was purchased from E. Merck (Mumbai, India).
Apparatus
A Series HPLC system PEAK LC7000 isocratic HPLC with PEAK 7000 delivery system. Rheodyne
manual sample injector with switch (77251),Analytical column Chromosil C18. 250×4.6mm, Electronic
balance-DENVER (SI234), A manual Rheodyne injector with a 20 μl loop was used for the injection of
sample., PEAK LC software were used. UV 2301 SPECOPHOTOMETER was used to determine the
wavelength of maximum absorbance

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Determination of wavelength of maximum absorbance
The standard solutions of Esomeprazole were scanned in U.V range against mobile phase as a
blank. Esomeprazole showed maximum absorbance at 272 nm. So the wavelength selected for the
determination of Esomeprazole was 272 nm.
Chromatographic equipment and conditions
The development and validation of the assay was performed on A Series 200 HPLC system PEAK
LC7000 isocratic HPLC with PEAK 7000 delivery system. Rheodyne manual sample injector with switch
(77251),Analytical column Chromosil 100-5 C18. 250×4.6mm, , manual injector rheodyne valve) with
20μL fixed loop, PEAK LC software were used.
The mobile phase consisted of a Methanol, Acetonitrile Tetrahydrofuran 60:30:10 (v/v). Injections
were carried out using a 20 μl loop at room temperature (20 + 2 °C) and the flow rate was 0.8 ml/min.
Detection was performed at 272 nm with 10 min runtime.
Standard and sample solutions
A 10 mg amount of Esomeprazole reference substance was accurately weighed, dissolved in10ml
mobile phase in a 10 ml volumetric flask to obtain 1000 ppm concentrated solution. From standard
solution by the serial dilution we prepared required concentrations of 100ppm.3ml of above sample was
taken and diluted to 10ml using mobile phase. A composite of 20 tablets was prepared by grinding them
to a fine, uniform size powder. 10 mg of Esomeprazole was accurately weighed and quantitatively
transferred into a 100 ml volumetric flask. Approximately 26 ml mobile phase were added and the
solution was sonicated for 15 min. The flask was filled to volume with mobile phase, and mixed. After
filtration, an amount of the solution was diluted with mobile phase to a concentration of 30 μg/ml.
Method validation
Method validation was performed following ICH specifications for specificity, range of linearity,
accuracy, precision and robustness
RESULTS AND DISCUSSION
System Suitability
Having optimized the efficiency of a chromatographic separation the quality of the
chromatography was monitored by applying the following system suitability tests: capacity factor, tailing
factor and theoretical plates. The system suitability method acceptance criteria set in each validation run
were: capacity factor >2.0, tailing factor ≤2.0 and theoretical plates >2000 13. In all cases, the relative
standard deviation (R.S.D) for the analytic peak area for two consecutive injections was < 2.0%. A

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chromatogram obtained from reference substance solution is presented. System suitability parameters
were shown in Table.1. Standard chromatogram was given in Figure.2
Table.1 System suitability parameters
Mobile phase Methanol,Acetonitrile:Tetrahydrofuran
60:30:10 (v/v)
Pump mode Isocratic
pH 5.8
Diluents Mobile phase
Column Zodiac C18 column (250 X 4.6 mm, 5μ)
Column Temp Ambient
Wavelength 272nm
Injection Volume 20 μl
Flow rate 0.8 ml/min
Run time 10 minutes
Retention Time 6.05 minutes

Figure.2: Standard solution chromatogram
Range of linearity
Standard curves were constructed daily, for three consecutive days, using six standard
concentrations in a range of 10, 20, 30, 40, 50,60μg/ml. for Esomeprazole. The linearity of peak area
responses versus concentrations was demonstrated by linear least square regression analysis. The linear
regression equation was y = 42598 + 6031x (r= 0.997). Linearity values can shown in Table: 2
Precision
To study precision, six replicate standard solutions of Esomeprazole(30 ppm) were prepared and
analyzed using the proposed method. The percent relative standard deviation (% RSD) for peak
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responses was calculated and it was found to be 0.89which is well within the acceptance criteria of not
more than 2.0%. Results of system precision studies are shown in Table.
Table.2: Linearity results
Concentration of Esomeprazole peak area
Level In ppm
Level - 1 10 65149
Level - 2 20 154690
Level - 3 30 254613
Level - 4 40 327894
Level - 5 50 399456
Level - 6 60 49675
Slope 8328
Range:10ppm Intercept 7214
to 60ppm Correlation coefficient 0.9988

600000

500000

400000
PEAK AREA

300000

200000

100000

0
0 10 20 30 40 50 60 70
-100000
SAMPLE CONCENTRATION

Graph.1: Linearity graph
Precision Results for Esomeprazole:
Table.3: Precision Results
Conc. (in ppm) Injection No. Intraday Interday
1 254932 254112
2 254614 254327
3 254032 254918
30
4 254516 254716
5 254739 254510
6 254419 254403
Limit of Detection and Limit of Quantification:
To determine the Limit of Detection (LOD) sample was dissolved by using Mobile phase and
injected until peak was disappeared. After 0.02 ppm dilution Peak was not clearly observed, based on
which 0.02 ppm is considered as Limit of Detection and Limit of Quantification is 0.06 ppm.
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Table.4: LOD and LOQ results
Parameter Measured Value
Limit of Quantification 0.06 ppm
Limit of Detection 0.02 ppm
Robustness
Typical variations in liquid chromatography conditions were used to evaluate the robustness of
the assay method. In this study, the chromatographic parameters monitored were retention time, area,
capacity factor, tailing factor and theoretical plates. The robustness acceptance criteria set in the
validation were the same established on system suitability test describe above.
Table.5: Robustness results
S.NO PARAMETER CONDITION AREA % OF CHANGE
1 Standard Standard conditions 254613 100%
2 Mobile phase Methanol 80%,ACN 10%,THF 10%. 254562 99.97%
3 Mobile phase pH 5.6 254179 99.82%
4 Wavelength 270 nm 254833 100%
Recovery
Recover test was performed at 3 different concentrations i.e. 20ppm,40ppm,60ppm. Results are
given in table.6
Table.6: Recovery results
Recovery Conc of sample Recovery % of recovery
50% 20PPM 19.85 99.25
100% 40PPM 40.02 100.05
150% 60PPM 59.95 99.91

Figure.3: Formulation sample
Table.7: Assay results
S.NO Formulation Dosage Sample conc Drug estimated % of Drug Estimated in Tablet
1 RACIPER 40 mg 40 ppm 39.95ppm 99.8

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CONCLUSION
The proposed method for the assay of Esomeprazole in formulations is very simple and rapid. It
should be emphasized it is isocratic. The method was validated for specificity, linearity, precision,
accuracy and robustness.From our validated reports we are concluded that this method may use for
analysis of Esomeprazole in formulations.
REFERENCES
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