INTRODUCTION:

Domperidone is a synthetic benzimidazole compound that acts as a dopamine D₂-receptor antagonist. Chemically,it is 5-chloro-1-[1-(2,3-dihydro-2-oxo-1H-benzimidazol-2-one¹.It is used as an antiemetic and antinauseant.It is also used as a prokinetic agent for treatment of upper gastrointestinal motility disorders^2,3.It is rapidly absorbed from the stomach and the upper part of GIT by active transport, after oral administration⁴.The recommended dosage for Domperidone is10 mg per day. It is official in British pharmacopoeia and European pharmacopoeia where non-aqueous titration is the official method of assay^5,6.Literature survey reveals that several methods like Spectrophotometry, HPLC, HPTLC and LC-MS were reported for the estimation of Domperidone in combination with other drugs as well as in biological fluids^7-22. Recently, we reported spectrophotometric method for the estimation of Domperidone in bulk and pharmaceutical formulation²³. The review of the literature revealed that no method is yet reported for the estimation of Domperidone in bulk and pharmaceutical formulation by RP- HPLC.

MATERIALS AND METHODS:

Standard sample of Domperidone was obtained from Dr.Reddy’s labs, Hyderabad, India. HPLC grade Acetonitrile, Methanol and AR grade potassium dihydrogen phosphate were procured from E-Merck. Double distilled water was prepared in our laboratory. Fixed dose tablet formulations Domstal and Vomistop containing domperidone (10mg/each tablet) were obtained from local market.

INSTRUMENTS USED:

Digital balance (Essae), pH meter (Elico), Ultrasonicator (Optics technologies), Millipore solvent filtration unit (Bros Scientifics),UV-detector, HPLC system (Shimadzu LC-20AT Prominence solvent delivery system), Analytical column-Phenomonex-Gemini, C- 18 (250x4.6mm id,5µ) were the instruments used in the present study.

RP-HPLC ASSAY METHOD:

The standard solutions of Domperidone, Domstal and Vomistop each containing 200 mcg/ml were prepared. From these solutions, 20µl were injected and chromatograph.

The mobile phase consisting of potassium dihydrogen phosphate buffer-acetonitrile-methanol (40:30:30) was pumped at a flow rate 1ml per min, the detection was monitored at 288 nm and the run time was 7 minutes.

Fig no 1: Chromatogram of Domperidone standard.

Fig no : 2 Chromatogram of Domperidone in DOMSTAL.

METHOD VALIDATION:^{24- 26}

It is defined as the process of proving that an analytical method is acceptable for its intended use. It proves that the method developed is specific, linear, precise, accurate and sensitive. The different parameters of analytical method validation are discussed below.

Accuracy:

It is the measure of exactness of an analytical method, or the closeness of agreement between the value which is accepted either as a conventional true value or an accepted reference value and the value found. Accuracy is evaluated by analyzing synthetic mixtures spiked with known quantities of components. For the quantitation of impurities, accuracy is determined by analyzing samples (drug substance or drug product) spiked with known amounts of impurities. To document accuracy²⁷the ICH guideline on methodology recommends collecting data from a minimum of nine determinations over a minimum of three concentration levels covering the specified range, for example, three concentrations, three replicates each. The data should be reported as the percent recovery of the known, added amount, or as the difference between the mean and true value with confidence intervals.

Table-1: Recovery of Domperidone in commercial tablets :

Formulation

Label content (mg/tablet)

Mean amount found( mg )

Mean

(%)

Standard deviation found

Domstal

Vomistop

9.80

10.15

98.0

101.5

0.64

0.58

Precision:²⁸

Precision is defined as the measure of the degree of repeatability and reproducibility of an analytical method under normal operation and expressed as the percent relative standard deviation for a statistically significant number of samples. According to ICH, it should be performed for repeatability, intermediate precision and reproducibility.

i) System precision:

The first type of precision study is instrument precision or injection repeatability. Repeatability should be assessed using a minimum of 9 determinations covering the specified range for the procedure (ex:3 concentrations/3 replicates each or a minimum of 6 determinations at 100% of the test concentration).

ii)Method precision:

The datas were obtained by repeatedly analyzing, in one laboratory on one day, aliquots of homogenous sample ,each of which independently prepared according to the following procedure.

Linearity:²⁹

It is the ability of the method to elicit test results that are directly proportional to analyte concentration with in a given range.

Ruggedness:³⁰

It is the degree of reproducibility of the results obtained under a variety of conditions, expressed as %RSD. An ICH chooses instead to cover the topic of ruggedness as part of precision (reproducibility).

Stability in analytical solution:

A sample solution containing 100 ppm of Standard Domperidone, Domstal and Vomistop was prepared and kept at room temperature and analyzed initially at different time intervals. As the cumulative RSD up to 12 hours meet the acceptance criteria , it is concluded that sample is stable in analytical solution for atleast 12 hours at room temperature.

Table-2: Standard validation protocol:

S. No

Parameters

Experiment

Acceptance Criteria

System precision

Method precision

Linearity

Ruggedness

Six replicate injections of standard solution

Sample of one batch prepared and analysed separately six times as per the method.

Linearity to be performed in the range of about 70-130% of standard/sample solution.

Sample of the same batch,prepared and analysed separately six times by two different instruments,using to different columns on different days.

RSD should not be more than 1%.

Correlation coefficient should not be less than 0.99.

Overall RSD should not be more than 2%.

RESULTS AND DISCUSSION:

The Chromatogram obtained for Domperidone (Standard),Domstal and Vomistop were shown in the figures 1,2 and3. Recovery of Domperidone in commercial tablets was shown in Table 1. Then the HPLC assay method was validated .The standard validation protocol was shown in Table 2. The validation results are as follows.

System Precision:

The RSD value was found to be 0.24,0.28 and 0.36 for Domperidone standard ,Domstal and Vomistop respectively and the values were found to be present within the acceptable limits. The order of system precision is as follows :Domperidone standard>Domstal>Vomistop.

Method precision:

The RSD value was found to be 0.38,0.42 and 0.44 for Domperidone standard, Vomistop and Domstal respectively and the values were found to be present within the accepatable limits. The order of method precision is as follows: Domperidone standard > Vomistop > Domstal.

Linearity:

The correlation coefficient value was found to be 0.9994,0.9996, and 0.9997 for Domperidone standard, Domstal and Vomistop respectively and the values were found to be present within the accepatable limits. The order of linearity is as follows:Domperidone standard>Domstal>Vomistop.

CONCLUSION:

From the above experimental data and validation results, the HPLC assay method for the estimation of Domperidone in bulk and pharmaceutical formulation is said to be rapid,simple,sensitive,precise,accurate and reliable for routine analysis in research institutions and quality control department in pharmaceutical industries.

ACKNOWLEDGEMENTS:

The authors are thankful to M/S Dr.Reddy’s laboratories private limited,Hyderabad,India, for providing Domperidone sample and Smt.P.Sulochana, , M.A., B.Ed., L.L.B, Correspondent,Sri Padmavathi Group of institutions,Tirupati for providing facilities to carry out this work.

Fig. no: 3 Chromatogram of Domperidone in VOMISTOP.

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Received on 03.11.2009 Modified on 09.11.2009

Asian J. Research Chem. 2(4):Oct.-Dec. 2009 page 561-564