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R.A Tamboli, V.C. Chauhan, M.M. Pathan, S.K. Tirgar, D.A. Shah, R.R. Parmar
APMC College of Pharmaceutical Education and Research,
Motipura, Himmatngar, Gujarat

A specific,  accurate,  precise  and  reproducible  RP-HPLC  method  has  been  developed  and subsequently validated for the simultaneous determination of Ilaprazole and Domperidone  in pharmaceutical dosage form. The proposed HPLC method utilizes hypersil (Thermo scientific) C18 column (250 mm × 4.6 mm id, 5 μm particle size), and mobile phase consisting of methanol:phosphate buffer (40:60) and pH adjusted to 4.0 with 0.1M glacial acetic acid at a flow rate of 1.0 mL/min. Quantitation was achieved with UV detection at 230 nm based on peak area with linear calibration curves at concentration ranges 5-15 μg/ml for Ilaprazole and 15-45 μg/ml for Domperidone. The retention time of  Ilaprazole and Domperidone were found to be 3.433 min and 5.860 min respectively. The method was validated in terms of accuracy,  precision,  linearity,  limits  of detection,  limits  of  quantitation  and  robustness. This method has been successively applied to marketed formulation and no interference from the formulation excipients was found.


PharmaTutor (ISSN: 2347 - 7881)

Volume 2, Issue 7

Received On: 30/04/2014; Accepted On: 07/05/2014; Published On: 01/07/2014

How to cite this article: RA Tamboli, VC Chauhan, MM Pathan, SK Tirgar, DA Shah, RR Parmar; Development and Validation of RP-HPLC Method for Simultaneous Estimation of Ilaprazole and Domperidone in Pharmaceutical Dosage Form; PharmaTutor; 2014; 2(7); 149-156

Ilaprazole is a proton pump inhibitor(PPI) used in the treatment of dyspepsia, Peptic ulcer disease (PUD), gastroesophageal reflux disease (GORD/GERD) and duodenal ulcer. It is available in strengths of 5, 10, and 20 mg. Clinical studies show that Ilaprazole is at least as potent a PPI as omeprazole when taken in equivalent doses. Studies also showed that Ilaprazole significantly prevented the development of reflux oesophagitis.[1].

Ilaprazole is chemically 2-[(RS)-[(4-methoxy-3-methylpyridin-2yl)methyl]sulfinyl]-5-(1H-pyrrol-1-yl)-1H-benzimidazole[3].

Fig.1.Chemical structure of Ilaprazole[4]

Domperidone chemically is 5-chloro-1-{1-[3-(2-oxo-2,3-dihydro-1H-1,3-benzodiazol-1-yl)propyl]piperidin-4-yl}-2,3-dihydro-1H-1,3-benzodiazol-2-one[7]. Fig.2

Domperidone acts as a gastrointestinal emptying (delayed) adjunct and peristaltic stimulant. The gastroprokinetic properties of domperidone are related to its peripheral dopamine receptor blocking properties. Domperidone facilitates gastric emptying and decreases small bowel transit time by increasing esophageal and gastric peristalsis and by lowering esophageal sphincter pressure. Antiemetic: The antiemetic properties of domperidone are related to its dopamine receptor blocking activity at both the chemoreceptor trigger zone and at the gastric level. It has strong affinities for the D2 and D3 dopamine receptors, which are found in the chemoreceptor trigger zone, located just outside the blood brain barrier, which - among others - regulates nausea and vomiting.[7]

Fig.2.Chemical structure of Domperidone[7]

Combination of Ilaprazole and Domperidone are used in treatment of peptic ulcer and gastroesophageal reflux disease (GORD/GERD) and duodenal ulcer.In the literature survey it was found that Ilaprazole and Domperidone were estimatedindividualy or in combination with other drugs by UV, HPLC, HPTLCSpectrofluorimethods[8-22]. But no method has been found for simultaneous estimation of Ilaprazole and Domperidone by chromatographic method. In the view of the need in the industry for routine analysis of Ilaprazole and Domperidone in formulation, attempts are being made to develop simple and accurate RP-HPLC method for simultaneous estimation of Ilaprazole and Domperidone and extend it for their determination in formulation.


RP–HPLC instrument equipped with SPD-20 AT UV-Visible detector, (LC-20AT, Shimadzu), Rheodyne injector (20 μl Capacity), BDS hypersil (Thermo scientific) C18 column (250 mm × 4.6 mm, 5 μ particle size) and Spinchrom software was used.

Chemicals and reagents
Reference standard of ILA and DOM were obtained from Montage laboratories PVT. LTD., Himatnagar. Methanol and used was of HPLC grade and Phophatebuffer(pH 4.0) and all other reagent were of AR grade.

Preparation of standard and test solutions
Preparation of mobile phase

Mobile phase were prepared by mixing of 400 ml of  methanol with 600 ml of phosphate buffer, whose pH was  adjusted to pH 4.0 by addition of glacial acetic acid. The mobile phase prepared was degassed by ultrasonication for 20 min, so as to avoid the disturbances caused by dissolved gases. The degassed mobile phase was filtered through 0.45 μ filters to avoid the column clogging due to smaller particles.

Preparation of standard stock solutions
An accurately weighed quantity of ILA(10 mg) and DOM(30 mg) were transferred to a 100 ml volumetric flask and dissolved and diluted to the mark with mobile phase to obtain standard solution having concentration of ILA (100 μg/ml) and DOM (300μg/ml).

Preparation of solutions for calibrationcurve
The calibration curves were plotted over the concentration range 5-15 μg/ml for ILA and 15-45μg/ml for DOM. From the stock solution 100 μg/ml of ILA and 300 μg/ml of  DOMprepared. From these working solutions of ILA and DOM (0.5 ml, 0.75 ml, 1.0 ml, 1.25 ml, 1.5 ml and 0.5 ml, 0.75 ml, 1.0 ml, 1.25 ml, 1.5 ml) were transferred to a series of 10 ml of volumetric flasks and diluted to the mark with mobile phase. Aliquots (20 μL) of each solution were injected under the operating chromatographic conditions described above.

Preparation of sample solution
Take quantity equivalent to 10 mg ILA and 30 mg DOM was transferred to 100 ml volumetric flask in Mobile Phase .The solution was filtered through whatman filter paper No. 41 and the volume was adjusted up to the mark with Mobile Phase. From this 1 ml of solution is diluted to 10 ml with the help of mobile phase to get final concentration of 10 μg/ml ILA and 30 μg/ml DOM.

The developed method was validated according to ICH guidelines. To check the system performance, the system suitability parameters were measured. System precision was determined on six replicate injections of standard preparations. Number of theoretical plates and asymmetry were measured.

Linearity was performed with five concentrations ranging from 5-15μg/ml and 15-45 μg/ml for ILA and DOM respectively. The peak areas versus concentration of drug were plotted and a linear least-square regression analysis was conducted to determine the slope, intercept and correlation coefficient (r) to demonstrate the linearity of the method.

The limit of detection (LOD) and limit ofquantitation (LOQ)
LOD and LOQ of ILA and DOM were calculated using the following equations as per International Conference on Harmonization (ICH) guidelines.
LOD = 3.3 × σ/S
LOQ = 10 × σ/S

Where σ = the standard deviation of the response
S = Slope of calibration curve.

The intraday and interday precision of the proposed method was determined by analyzing the corresponding responses 3 times on the same day and on 3 different days 3 different concentrations of sample solutions of ILA (5μg/ml, 10 μg/ml and 15 μg/ml) and DOM (15 μg/ml, 30 μg/ml and 45 μg/ml). Percentage relative standard deviation (RSD) was calculated

Accuracy was performed by adding known amounts of ILA and DOM to the pre-analysed marketed formulation and then comparing the added concentration with the found concentration. Three levels of solutions were made which correspond to 80, 100 and 120% of the nominal analytical concentration (5 μg/ml for ILA and 15μg/ml for DOM). Each level was prepared in triplicate. The percentage recoveries of ILA and DOM at each level were determined. The mean recoveries and the relative standard deviation were then calculated.

The robustness of the method was evaluated by assaying the test solutions after slight but deliberate changes in the analytical conditions i.e. flow rate (± 0.2 ml/ min), proportion of buffer and methanol (62:38 and 58:42 v/v), and pH of buffer (± 0.2).



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