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Validated RP-HPLC Method for Estimation of Ranitidine Hydrochloride,

Domperidone and Naproxen in Solid Dosage Form

Article · July 2011

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Asian J. Pharm. Ana. 2011; Vol. 1: Issue 3, Pg 59-63 [AJPAna.]

ISSN- 2231–5667 (Print) www.asianpharmaonline.org

ISSN- 2231–5675 (Online) 0974-3618

RESEARCH ARTICLE
Validated RP-HPLC Method for Estimation of Ranitidine Hydrochloride,
Domperidone and Naproxen in Solid Dosage Form

Md. Ahsanul Haque1, Mohammad Shahriar1, Most. Nazma Parvin2 and S. M. Ashraful Islam1*
1
Department of Pharmacy, University of Asia Pacific, Dhanmondi, Dhaka-1209, Bangladesh
2
Department of Pharmacy, Stamford University Bangladesh, 51, Siddeswari Road, Dhaka-1217, Bangladesh
*Corresponding Author E-mail: ashraf@uap-bd.edu

ABSTRACT:
In the present study, a simple, sensitive and specific liquid chromatography (RP-HPLC) method has been developed
and validated for the quantification of ranitidine hydrochloride, domperidone and naproxen in solid dosage form. A
shim-pack CLC-ODS column (250 mm X 4.6 mm, 5 and a mobile phase constituting 0.1 M orthophosphoric acid
solution (pH 3.0): methanol (35:65 v/v) were used. The flow rate was 1.0 ml/min and detection was carried by using
ultraviolet (UV) detector at a wavelength of 280 nm. The retention times of ranitidine hydrochloride, domperidone and
naproxen were 2.702 min, 3.666 min and 9.842 min respectively. The peaks were well separated (resolution 4.55 and
20.2). The calibration curves were linear over the concentration range of 80% to 120% of target concentration (R2 >
0.999 for ranitidine and naproxen and 0.998 for domperidone). The method is accurate with 99.5% to 100.04%
recovery (% RSD < 1.23). The proposed method was successfully applied for the estimation of ranitidine
hydrochloride, domperidone and naproxen and potency was found within limit. Therefore, this method can be used for
the analysis of ranitidine hydrochloride, domperidone and naproxen in single or combine dosage form.

KEYWORDS: Ranitidine hydrochloride, domperidone, naproxen, method validation, HPLC, quantitative analysis.

INTRODUCTION:
Ranitidine hydrochloride is a H2- receptor antagonist and is
widely used for the short-term treatment of duodenal ulcer
and for the management of hypersecretory conditions.
Chemically Ranitidine HCl is N-[2-[[[-5-[(Dimethylamino)
methyl]-2-furanyl] methyl] thio] ethyl]-N- methyl-2-nitro-1,
1-ethenediamine hydrochloride.
Domperidone is a D2 receptor antagonist. It increases
gastrointestinal peristalsis and motility that prevent reflux
esophagitis and it is used to prevent nausea and vomiting.
Chemically domperidone is 5- chloro- 1- [1- [3- (2- oxo- 2,
3- dihydro- 1H-benzimidazol- 1- yl) propyl]- piperidin- 4-
yl]- 1, 3- dihydro- 2H benzimidazol- 2- one.
Naproxen is a well-known non-steroidal anti-inflammatory
drug which is clinically used in treatment of rheumatoid
arthritis and other painful musculoskeletal disorders. It
works by inhibiting both the COX-1 and COX-2 enzymes.
Chemically naproxen is 2-Naphthaleneacetic acid, 6-
methoxy- -methyl-,(s)-(+)-(s)-6-Methoxy- -methyl-2-
naphthaleneacetic acid.
These three drugs are widely prescribed either in single or
in combination for various disease conditions. Naproxen
and ranitidine combination is widely prescribed by the
physicians in order to avoid NSAID induced ulcers. On the
other hand domperidone is widely prescribed with
ranitidine to control reflux esophagitis, nausea and gastritis.
So it is highly needed to develop a simple method for
simultaneous determination of these drugs in combination
dosage form.

All the three drugs, ranitidine hydrochloride, domperidone

and naproxen, are official in BP1. But only ranitidine
Received on 22.08.2011 Accepted on 26.08.2011 hydrochloride and naproxen are official in USP2. BP
© Asian Pharma Press All Right Reserved describes HPLC method for ranitidine tablet and UV
Asian J. Pharm. Ana. 1(3): July-Sept. 2011; Page 59-63 method for naproxen tablet. On the other hand, USP

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Asian J. Pharm. Ana. 2011; Vol. 1: Issue 3, Pg 59-63 [AJPAna.]

describes HPLC method both for ranitidine and naproxen volumetric flasks and diluted up to the mark with mobile
tablet. But none describes any method for simultaneous phase to get five different concentrations (80%, 90%, 100%
determination of ranitidine hydrochloride, domperidone and 110% and 120% of target concentration). Solution
naproxen. Literature survey reveals that HPLC methods containing mixture of ranitidine, domperidone and
have been reported for the assay of ranitidine naproxen of five different concentrations (80%, 90%, 100%
hydrochloride3, domperidone4 and naproxen5 in individual 110% and 120% of target concentration) were prepared by
formulations. HPLC methods for the analysis of these three diluting aliquots of standard solutions of ranitidine
drugs in combination with other drugs are also reported6-9. domperidone and naproxen in 100 volumetric flasks.
But not a single method is reported for the simultaneous
determination of ranitidine hydrochloride, domperidone and Preparation of sample solution:
naproxen. So, the present work was undertaken with the Average weight of ranitidine tablet 150 mg, domperidone
aim to develop and validate an economic, rapid reversed- tablet 10 mg and naproxen tablet 250 mg were calculated.
phase high performance liquid chromatographic method Then the tablets were grinded separately to a fine powder
with high resolution according to ICH and Global Quality with the help of mortar and pestle. Then, powder containing
guideline10-11. The method will be helpful to determine 75 mg ranitidine, 5 mg domperidone and 125 mg naproxen
these three drugs form either single or combine was transferred to a volumetric flask, dissolved in mobile
pharmaceutical dosage form. phase, shaken for about 10 minutes and filtered through
filter paper. The filtered solution was further diluted in the
MATERIALS AND METHOD: mobile phase to make the final concentration of working
Materials: sample equivalent to 100% of target concentration.
Ranitidine hydrochloride and domperidone were provided
by General Pharmaceuticals limited Dhaka, Bangladesh and Validation of HPLC method:
naproxen was a gift from Eskayef Bangladesh Ltd. Present study was conducted to obtain a new, affordable,
Methanol was of HPLC grade and was purchased from E. cost-effective and convenient method for HPLC
Merck, Darmstadt, Germany. Orthophosphoric acid and determination of ranitidine hydrochloride, domperidone and
other reagents were of analytical-reagent grade and naproxen in solid dosage form. The method was validated
purchased from E. Merck, Darmstadt, Germany. Water was for the parameters like system suitability, selectivity,
deionised and double distilled. Ranitidine tablet 150 mg, linearity, accuracy, precision and robustness.
domperidone tablet 10 mg and naproxen tablet 250 mg were
purchased from local drug store in Dhaka city after The system suitability was assessed by six replicate
checking their manufacturing license numbers, batch analyses of standard solution at a 100% level to verify the
numbers, production and expiry dates. resolution and reproducibility of the chromatographic
system. This method was evaluated by analyzing the
Instrumentation and chromatographic conditions: repeatability of retention time, tailing factor, theoretical
A Shimadzu (Japan) HPLC system consisting of a CMB-20 plates (Tangent) of the column and resolution between the
Alite system controller, two LC-20AT pumps, SIL-20A drug peaks.
auto-sampler and CTO-10ASVP column oven were used.
Ultraviolet detection was achieved at 280 nm with a SPD- To determine the selectivity of the method, standard
20A UV-VIS detector (Shimadzu, Japan). The drug samples of ranitidine, domperidone and naproxen were
analyses data were acquired and processed using LC injected first. Then solution containing mixed components,
solution (Version 1.3, Shimadzu, Japan) software running commercial product, blank and excipients solution were run
under Windows XP on a Pentium PC. The mobile phase, a in the instrument one after another. The chromatograms
mixture of 0.1 M orthophosphoric acid solution (pH 3.0): were analyzed for retention time, peak area and peak shape
methanol (35:65 v/v) pumped at a flow rate of 1.0 ml/min to determine selectivity of the method.
through the column (C18; 250 mm X 4.6 mm, 5 shim-pack,
Japan) at 300C. The mobile phase was filtered through a The linearity of an analytical method is its ability to elicit
0.2 nylon membrane filter and degassed prior to use under that test results are proportional to the concentration of
vacuum. Elusions were analyzed by UV detector at a analyte in samples within a given range. This was
sensitivity of 0.0001. determined by means of calibration graph using increasing
amounts of standard solutions (80%, 90%, 100%, 110%
Preparation of standard solution: and 120% of target concentration). These standards were
Stock solution of ranitidine, domperidone and naproxen tested six times in agreement to 10
the International
were prepared as per their dose ratio. Stock solution of Conference on Harmonization (ICH) . Calibration curves
ranitidine (600µg/ml), domperidone (200µg/ml) and were constructed and the proposed method was evaluated
naproxen (1000µg/ml) were prepared by dissolving 60 mg by its correlation coefficient and intercept value,
ranitidine (as ranitidine hydrochloride), 20 mg domperidone calculated in the corresponding statistical study (ANOVA)
and 100 mg naproxen in 100 ml mobile phase separately. (p < 0.05).
Several aliquots of standard solutions of ranitidine,
domperidone and naproxen were taken in different 100 ml

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Asian J. Pharm. Ana. 2011; Vol. 1: Issue 3, Pg 59-63 [AJPAna.]

Table-1: Results of system suitability study

Parameters Ranitidine Domperidone Naproxen
Average SD %RSD Average SD %RSD Average SD %RSD
Retention time 2.702 0.001 0.033 3.666 0.001 0.024 9.842 0.018 0.179
Area 1046302.33 3218.493 0.308 108585.333 468.352 0.431 1682585 103.811 0.006
Theoretical plates 2752.333 3.141 0.114 4596.000 10.733 0.234 10106 79.679 0.788
Tailing factor 1.175 0.003 0.288 1.322 0.004 0.295 1.702 0.059 3.466
Resolution 4.55 0.026 0.577 20.2 0.022 0.110

Characteristic parameters for regression equation (y = a + Wave length for UV detection was determined by scanning
bx) of the HPLC method were obtained by least squares individual and combined solution of ranitidine
treatment of the results and these parameters were used to hydrochloride, domperidone and naproxen. Then HPLC
confirm the good linearity of the method. analysis of individual and combined standard was measured
at 228, 240, 280, 314, and 332 nm. Finally, all the analysis
Accuracy indicates the deviation between the mean value were done at 280 nm as at this wavelength, all the three
found and the true value. Accuracy was determined by drugs absorbed light better and peaks could be
means of recovery experiments, by the addition of active distinguished properly.
drugs to placebo formulations. The accuracy was calculated
from the test results as the percentage of the analyte Method validation:
recovered by the assay. The experiment was carried out according to the official
specifications of USP, ICH- 1995, and Global Quality
The precision of the method was investigated with respect Guidelines.2, 10-11
to repeatability, ruggedness (intermediate precision) and
reproducibility (by means of an inter laboratory trial). System Suitability:
Repeatability was determined by performing four repeated The system suitability tests were carried out to evaluate the
analysis of the three standard solutions (90%, 100% and resolution and reproducibility of the system for the analysis.
110% of target concentration) on the same day, under the Table 1 represents system suitability results of this method.
same experimental conditions. Ruggedness (intermediate The system is found suitable in respect of retention time (%
precision) of the method was assessed by carrying out the RSD 0.001-0.179) mean theoretical plate count (% RSD
analysis of standard solutions on three different days (inter- 0.114-0.788) and resolution between peaks.
day) in the same laboratory. For reproducibility the
procedure repeated in the Quality Control Laboratory of Selectivity:
another lab. The relative standard deviation (% RSD) was Selectivity is the ability of the method to assess the
determined in order to assess the precision of the method. analyte in the presence of excipients, impurities,
The robustness of the method was assessed by altering the degraded products, matrix etc. Peaks from individual
some experimental conditions such as, by changing the flow sample solutions and mixed sample solution were on
rate from 0.9 to1.1 ml/min, amount of methanol (63% to same time (Figure 1). On the other hand no other peaks
67%) the temperature of the column (28 °C to 32 °C) and other than drugs were found within 20 min run time
PH (2.9-3.1) of the mobile phase. from the commercial products. Blank and excipients
did not change the retention time or interfere the
RESULTS AND DISCUSSION: analysis results. So the method is highly selective for
Methods development and optimization: ranitidine hydrochloride, domperidone and naproxen.
This isocratic mode method with UV detection was
developed for the determination of the ranitidine
hydrochloride, domperidone and naproxen. The mobile
phase was chosen after several trials with 0.1 M ortho
phosphoric acid and methanol in various proportions like
70: 30, 65:35, 50:50, 40: 60, 30:70 and 35:65 at different
pH values. When aqueous phase was increased in mobile
phase tailing factor and retention time of domperidone and
naproxen were also increased. So, mobile phase containing
0.1 M ortho phosphoric acid solution (pH 3.0) and methanol
(35:65 v/v,) was selected to achieve maximum separation
and sensitivity.

Different flow rates in between 0.50 to 2.0 ml /min were

studied. A flow rate of 1.0 ml /min gave an optimal signal
to noise ratio with a reasonable separation time. Figure 1.Chromatogram of combined (C) and single sample of
ranitidine (R), domperidone (D) and naproxen (N).

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Asian J. Pharm. Ana. 2011; Vol. 1: Issue 3, Pg 59-63 [AJPAna.]

Linearity: Accuracy and Precision:

Linearity of the method was evaluated from the correlation
coefficient of calibration curves that were constructed from
average peak area of drugs at different concentration level
(80%, 90%, 100%, 110% and 120%). Correlation
coefficient was 0.999 for ranitidine hydrochloride and
naproxen and 0.998 for domperidone (Table 2 and Figure
2).
Results of accuracy and precision (repeatability, ruggedness
and reproducibility) are summarized in table 2 along with
linearity results. Accuracy is generally assessed by
analyzing samples with known concentration and
comparing the measured value with the true value .The
measured value was obtained by recovery test. % recovery
was found 99.5% to 100.04% with % RSD value 0.37 to
1.23. All the results indicate that the method is highly
accurate.

The measurement for repeatability was done from 9.00 am

to 9.00 pm. Four determinations of three concentrations
across the intended range (90%, 100% and 110% of target
concentration) were included in the study. % RSD of peak
areas was calculated for various run. The method is highly
precise as % RSD of peak area was 0.35%.

Test results for ruggedness were obtained by analyzing

three concentrations (90%, 100% and 110% of target
concentration) with 4 runs over 3 days. The average peak
area obtained at different levels and different days indicate
that the method is precise. % RSD values were found
slightly higher for all the drugs in this study than
repeatability study. Samples analysis results in another lab
Figure 2.Calibration curve of ranitidine hydrochloride (R), (reproducibility) were also within limit (% RSD was less
domperidone (D) and naproxen (N). than 2%).

Table 2: Results of a linearity, accuracy and precision study

Validation parameters Ranitidine Domperidone Naproxen
Linearity (regression coefficient- R2 (mean±SD) 0.9997±0.0002 0.9980±0.0005 0.9993±0.0002
R 2) %RSD ** 0.021 0.056 0.021
(*Y = mX+C) Slope (mean±SD) 22987.87±38.79 33871.77±470.05 22782.91 ±78.77
Accuracy % Recovery 100.04 ± 0.37 100.01± 1.23 99.5± 0.52
%RSD 0.37 1.23 0.53
Precision (%RSD) Repeatability 0.35 0.27 0.095
Ruggedness 0.59 0.43 0.25
Reproducibility 1.32 0.94 0.83
*Y = mX+C; where Y = peak area, m = slope, X = concentration ( g/ml) and C = intercept.
**%RSD = Relative standard deviation = (Standard deviation X 100)/mean, R2 = Correlation coefficient

Table 3: Analysis of marketed tablet and new formulated capsule by proposed HPLC method
Run Commercial tablet New formulated capsule (ranitidine 150 mg, domperidone 10 mg and
No. naproxen 250 mg)
Ranitidine tab Domperidone tab Naproxen tab Ranitidine Domperidone Naproxen
(150 mg) (10 mg) (250 mg)
% potency
R-1 99.01 102,16 103.95 99.57 99.93 100.36
R-2 98.93 104.62 100.46 99.23 99.23 100.00
R-3 100.05 102.97 102.81 99.85 98.82 98.97
Average 99.33 103.80 102.41 99.55 99.33 99.78
SD 0.625 1.167 1.780 0.310 0.561 0.723
% RSD 0.629 1.124 1.738 0.312 0.565 0.725

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 Asian J. Pharm. Ana. 2011; Vol. 1: Issue 3, Pg 59-63
Robustness:
Robustness study was performed by making slight
variations in flow rate, amount of methanol, temperature
and pH of the mobile phase. No significant effect was
observed in the recovery of drugs. % recovery was 98% to
102%. On the other hand changes in retention time,
theoretical plate, resolution were also negligible. So we can
say that the method is robust.
Analysis of market products:
The proposed method was used to determine the potency of
commercially available tablets (Ranitidine tablet 150 mg,
domperidone tablet 10 mg and naproxen tab 250 mg).
Capsule containing 150 mg ranitidine, 10 mg domperidone
and 250 mg naproxen were prepared using common
excipients and analyzed. Three replicate determinations
(n=3) were carried out and the results are summarized in
Table 3.
CONCLUSION:
The validation study shows that the developed method is
accurate, rapid, precise, reproducible and inexpensive with
acceptable correlation co-efficient, RSD (%) and standard
deviations which make it versatile and valuable for
simultaneous determination of ranitidine hydrochloride,
domperidone and naproxen in bulk or pharmaceutical
dosage form (individual or combine). The advantages lie in
the simplicity of sample preparation and the low cost of
reagents used. The proposed method is simple and do not
involve laborious time-consuming sample preparation. So
this RP-HPLC method can be used in the quality control
department.
ACKNOWLEDGEMENTS:
The authors are thankful to General Pharmaceuticals limited
Bangladesh, for providing gift samples of ranitidine
hydrochloride and domperidone. Authors are also thankful
to Eskayef Bangladesh Ltd. for providing gift samples of
naproxen.
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