Simultaneous Estimation of Rabeprazole Sodium and
Domperidone Maleate in Bulk and Tablet Dosage Form by Reverse Phase High
Performance Liquid Chromatography
Jawed Akhtar1*, B Srivastava1,
SS Sukla2, Rajeev Chaturvedi1 and Uttam Singh Baghel1
1School of
Pharmaceutical Sciences, Jaipur National University, Jagatpura, Jaipur.
Rajasthan,
2Columbia
Institute of Pharmacy, Raipur, Chhattisgarh.
*Corresponding
Author E-mail: jawed_pharma@yahoo.com
ABSTRACT:
A selective and sensitive reverse phase high
performance liquid chromatography (RP-HPLC) method has been developed for the
separation and quantification of rabeprazole sodium and domperidone maleate in
tablet dosages form has been fist developed and validated. Quantification
carried out using HIQ Sil C18 column (25cm x 4.6mm i.d.) as a
stationary phase and mobile phase comprised of 50mM KH2PO4
and acetonitrile in proportion of 60:40 (v/v) with pH adjusted to 7 ±1 by using
5M potassium hydroxide. The flow rate was 1.0 ml/min and monitored at 285 nm.
The retention time for rabeprazole and domperidone were 6.707 and 8.002 min,
respectively. The method was validated in terms of linearity, precision,
accuracy, ruggedness, and specificity, limit of detection and limit of
quantification. The linearity (r2) and percentage recoveries of
rabeprazole and domperidone were 0.9986 and 100.02µg/ml and 0.9992 and 99.98 %
respectively. The proposed method is suitable for simultaneous determination of
rabeprazole and domperidone in tablet dosages form.
KEYWORDS:
RP-HPLC; Rabeprazole sodium; Domperidone maleate;
simultaneous determination.
1. INTRODUCTION:
Rabeprazole sodium is chemically
2-[[[4-(3-methoxypropoxy)-3-ethyl-2-pyridinyl]-methyl]
sulfinyl]-1H–benzimidazole sodium salt1. It is a newer proton pump Inhibiter used in the treatment
of ulcer, and Gastroesophageal reflux disease, compound that inhibit the H+
K ATPs. The drug is not yet official in any of the pharmacopoeia. A survey of
literature reveals that only HPLC2-5 (in plasma) methods have been
reported for the estimation of Rabeprazole sodium.
Domperidone
maleate is chemically 5-chloro-1- [1-[3-(2-oxo-2, 3-dihydro – 1H–benzimidazole-1-yl)
propy l]-pipridin-4-yl]-1, 3-dihydro-2H-benzimidazol -2-one6. It is
a dopamine antagonist used as an antiemetic for short-term treatment of nausea
and vomiting of various etiology7. It is official in British
Pharmacopoeia (BP)8. The method of analysis given in B.P by non-aqueous
titration. Methods such as HPLC9,10 and Spectrophotometric11
have been reported in the literature.
Even though
various methods reported in the literature for estimation of rabeprazole sodium
and domperidone maleate individual or in combination with other drugs no method
had been reported so far for simultaneous estimation of these two drugs using
HPLC in tablet dosage forms. The present study was aimed at the simultaneous
estimation of rabeprazole sodium and domperidone maleate by HPLC method. This
method was validated according to the ICH guidelines12.
2. EXPERIMENTAL:
2.1. Instrument:
A HPLC (LC-20AD, shimadzu, Japan) connected to computer
loaded with spinchrom chromatographic software. System was coupled with SPD-20A
prominence UV/Vis detector. All weights were taken on semi microbalance (Shimadzu
-AX -200 electronic balance)
2.2. Reagents and chemicals;
Rabeprazole sodium and Domperidone maleate were
obtained as gift sample from Torrent Pharmaceutical Pvt. Ltd., Ahmedabad (Gujarat). Pottasium dihydrogen phosphate, and potassium hydroxide were A.R. grade from
Merck chemicals Mumbai, India. Acetonitrile and milli Q water were HPLC
grade supplied by Merck chemicals Mumbai, India.
Table No.1: System
suitability
|
S. No
|
Parameter
|
Rabeprazole sodium
|
Domperidone maleate
|
|
1
|
Capacity factor
|
1
|
1
|
|
2
|
Theoretical Plate
|
6804
|
5587
|
|
3
|
Asymmetry of the peak
|
1.14
|
0.91
|
|
4
|
Retention time
|
8.002
|
6.958
|
|
5
|
Resolution
|
3.11
|
2.3.
Chromatographic conditions:
A HIQ- Sil
C18 (25cm x 4.6 mm, 5μ) column was used as the stationary
phase. The mobile phase comprised of 50mM KH2PO4 and acetonitrile
in proportion of 60:40 (v/v) with pH adjusted to 7 ±1 by using 5M potassium
hydroxide, was used. Injection volume was 20 μl and run time was 15min and
flow rate 1.0 ml/min. The column was maintained at ambient temperature and the
eluent detected at 285 nm.
2.4. Standard
preparation:
Standard
stock solution (1000μg) of Rabeprazole sodium and Domperidone maleate wear
prepared separately in mobile phase comprised of 50mM KH2PO4
and acetonitrile in proportion of 60:40 (v/v) with pH adjusted to 7 ±1 by using
5M potassium hydroxide. The Working standard solutions were prepared and
further diluted in mobile phase to contain a mixture of Rabeprazole sodium and
Domperidone maleate in over the linearity range from 2-100 µg/ml and 2-50 µg/ml
respectively.
2.5.
Sample preparation:
Twenty tablets, were weighed and finely powdered. A
quantity of powder equivalent to 20 mg of rabeprazole sodium and 10mg of
domperidone maleate was weighed and transferred to a 100 ml volumetric standard
flask and added 50 ml of mobile phase than sonicated for 15 min. The solution
was cooled and made up to the mark with mobile phase. The contents were mixed
thoroughly and filtered through a 0.45µ filter. 10µl of the sample was injected
in to HPLC system.
3. RESULTS AND DISCUSSION:
The proposed HPLC method
required fewer reagents and materials, and it is simple and less time
consuming. This method could be used in quality control test in pharmaceutical
industries. The chromatograms of Rabeprazole sodium and Domperidone maleate were
shown in (fig. 1).There was clear resolution between Rabeprazole sodium and
Domperidone maleate with retention time of 6.707 and 8.002 minutes
respectively.
3.1. Validation of the
method:
3.1.1. System suitability:
The Rt values, capacity
factor, resolution, theoretical plates/meter, and peak symmetry were calculated
for the standard solutions. The values obtained demonstrated the suitability of
the system for the analysis of the above drug combinations System suitability
parameters might be fall within ± 3% standard deviation range during routine
performance of the method. The summary of the system suitability results were
showed in the (table 1).
Figure 1: Typical chromatogram of Rabeprazole sodium
and Domperidone maleate
3.1.2. Linearity:
The response for the
detector was determined to be linear over the range of 2-100 µg/ml (2, 5, 10,
25, 50, 100) for Rabeprazole sodium and 2-50 µg/ml ( 2, 5, 10, 25, 50) for
Domperidone maleate. Each of the concentration was injected in duplicate to get
reproducible response. The calibration curve was plotted as concentration of
the respective drug versus the response at each level. The proposed method was
evaluated by its correlation coefficient and intercept value calculated in the
statistical study. They were represented by the correlation co-efficient in
(table 2).
3.1.3. Precision and
accuracy:
The accuracy of the method
was determined by recovery experiments. The recovery studies were carried out 6
times and the percentage recovery and % relative standard deviation was
calculated. From the data obtained, recoveries of standard drugs were found to
be accurate (table 2).The %RSD of interday and intraday precision obtained was
less than 2% for both the drugs. The intraday and interday precision of
Rabeprazole sodium was 0.1978 and 0.1783 and Domperidone maleate was 0.1729 and
0.2653 respectively. From the data obtained, the developed HPLC method was
found to be precise and accurate.
3.1.4. Specificity of the
method:
The chromatograms of the
rabeprazole sodium and domperidone maleate in standard and placebo were
recorded. In the chromatograms of the formulations, some additional peaks were
observed which may be due to excipients present in the formulations. These
peaks however did not interfere with the standard peaks, which demonstrate that
the assay method is specific. Furthermore, the purity of the peaks was studied
by peak purity studies. The results revealed that the peak is free from
interferences, which shows that the HPLC method is specific.
3.1.5. Detection and
Quantification limits:
The limit of detection (LOD)
and limit of quantification (LOQ) of the developed method determined by
injecting progressively low concentrations of the standard solutions using the
developed methods.
Table No.2: Summary
of analytical method validation
|
S. No
|
Parameters
|
Acceptance criteria
|
Rabeprazole sodium
|
Domperidone maleate
|
|
1
|
Linearity
|
r2 = 0.995 to 1.0
|
0.9986
|
0.9992
|
|
2
|
Specificity
|
No interference with placebo
|
Specific
|
Specific
|
|
3
|
Accuracy (Recovery studies)
|
Recovery : 98.0 to 102.0%
|
100.02%
|
99.98 %
|
|
4
|
Precision
|
|
Intraday (n=6)
|
RSD NMT 2.0%
|
0.1978
|
0.1729
|
|
Interday (n=6)
|
RSD NMT 2.0%
|
0.1783
|
0.2653
|
|
5
|
Robustness ( pH )
|
NMT  1%
|
0.2%
|
0.4%
|
|
6
|
Limit of detection (mg / ml)
|
………………..
|
0.18
|
0.56
|
|
7
|
Limit of quantitation (mg / ml)
|
………………..
|
1.31
|
1.57
|
The LOD is the lowest
concentration of the analyte that can be detected with signal to noise ratio
(1:3) and LOQ is the lowest concentration that can be quantified with
acceptable precision and accuracy with signal to noise ratio (1:10). The LOD of
rabeprazole sodium and domperidone maleate found to be 0.18µg/ml and 0.56µg/ml
respectively. The LOQ of rabeprazole sodium and domperidone maleate found to be
1.31µg/ml and 1.57µg/ml respectively.
3.1.6. Robustness:
The robustness of the method
was studied by deliberate changes in the method like percentage organic
content, alteration in pH of the mobile phase, changes in the wavelength. It
was observed that there was no marked changes in the chromatograms demonstrate
that the HPLC methods have developed are robust.
4. CONCLUSION:
This method is simple,
specific and easy to perform and requires short time to analyze the samples.
Low limit of quantification and limit of detection makes this method suitable
for use in quality control. This method enables simultaneous determination of
rabeprazole sodium and domperidone maleate because of good separation and
resolution of the chromatographic peaks. The method was found to be linear,
precise, accurate, rugged and robust.
5.
ACKNOWLEDGEMENTS:
The authors are thankful to
Torrent Pharmaceutical Pvt. Ltd. (Gujarat) for providing the gift samples of
rabeprazole sodium and domperidone maleate.
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