An Isocratic RP-HPLC Method for Simultaneous Analysis of Ilaprazole And Domperidone in Pharmaceutical Formulation


Tandel Jinal N.1*, Pumbhadiya Dilipkumar A.1, Chauhan Payal P1, Shah Samir K2

1Department of Quality Assurance, Sardar Patel College of Pharmacy, Gujarat Technological University, Bakrol, Gujarat

2Department of Pharmacology, Sardar Patel College of Pharmacy, Gujarat Technological University, Bakrol, Gujarat

*Corresponding Author E-mail:



A simple, sensitive and precise high performance liquid chromatographic method for the analysis of Ilaprazole (ILA) and Domperidone (DOM) in their combine dosage form has been developed and validated. The compounds were well separated an isocratically on a C18 Hyperchrom ODS-BP column (250 mm 4.6 mm, 5 µm) utilizing a mobile phase consisting of methanol: phosphate buffer pH 3.5 (35:65 % v/v,) at a flow rate of 1.0 ml/min with UV detection at 281.1 nm. The retention time of ILA and DOM was found to be 3.84 min and 6.55 min. respectively. The proposed method is linear over the concentration ranges 5–15 μg/ml and 15–45 μg/ml for ILA and DOM respectively. Accurate with 99.80-100.09 % recovery for ILA and 99.40-99.77 % recovery for DOM and precise (% RSD < 2 %).The LOD were 3.38 and 4.28 μg/ml and LOQ were 10.24 and 12.99 μg/ml for ILA and DOM respectively. The method has been used to determine potency of commercial product and potency was found within limit. The method is applicable  for the analysis of Ilaprazole and Domperidone in marketed formulation.


KEYWORDS: Domperidone, Ilaprazole, Validation, RP-HPLC., LUPILA-D.




Ilaprazole is a substituted benzimidazole which is antiulcerous compound known for decreasing gastric acid secretion. This compound, also known as proton pump inhibitor (PPI) is commonly indicated for the treatment of gastric ulcer, peptic ulcer, duodenal ulcers, erosive or ulcerative GERD (Gastro esophageal reflux disease), symptomatic GERD, pathological hypersecretory conditions.


The chemical structures of ilaprazole is [-[(4-methoxy-3-methyl-pyridin-2-yl) methylsulfinyl]-6-pyrrol-1-yl-1H. (Figure.1)(1)



Figure 1- Chemical Structures of Ilaprazole


Domperidone,5-chloro-1-[1-[3-(2,3-dihydro-2-oxo-1Hbenzimidazol-1-yl)propyl]-4-piperidinyl]-1, 3-dihydro-2Hbenzimidazol-2-one (Figure 2)(2) , is a potent dopamine antagonist used for treatment of nausea and vomiting . Domperidone does not cross the blood-brain barrier and therefore has fewer adverse CNS effects than other dopamine antagonists.(3)



Figure 2- Chemical Structures of Domperidone.


Literature survey reveals that many analytical methods have been reported for determination of ilaprazole  individually and in combination with other drug (4-6) and for the determination of Domperidone various methods like, UV(7-9), HPLC(10-11) Stability indicting HPLC(12) HPTLC(13) and LC/MS(14) No single method was reported for the combination of these two. So attempt was taken to develop and validate an economic, rapid reversed phase high performance liquid chromatographic method for the quality control of ILA and DOM in pharmaceutical preparations with lower solvent consumption along with the short analytical run time that leads to an environmentally friendly chromatographic procedure and will allow the analysis of a large number of samples in a short period of time. The method was validated and found to be simple, accurate, and          precise. (15)



Ilaprazole was procured from Lupin Ltd and domperidone was procured from Torrent pharmaceuticals, Gujarat. High Performance Liquid Chromatography (HPLC) grade acetonitrile, methanol and water were obtained from Chemdyes, Rajkot, Gujarat. Potassium dihydrogen phosphate, ortho Phaosphoric acid and methanol of Analytical Reagent (AR) grade were obtained from Chemdyes, Rajkot, Gujarat. Lupila- D capsules were purchased from local market in Gujarat.


Chromatographic conditions:

The instrument used was an Analytical Technologies with Rheodyne Injector, 2203 UV-Visible detector and the data recorded using Alchrome A 2000 software. Hyperchrome Octa DecylSilane (ODS) C18, (250mm × 4.6mm, 5µm) was used utilizing a mobile phase consisting of methanol: phosphate buffer (35:65, v/v, pH 3.5) at a flow rate of 1.0 ml/min with UV detection at 281.1 nm.


Preparation of standard stock solution:

10 mg and 30 mg of standard ilaprazole (ILA) and domperidone(DOM) was weighed accurately and transferred to two separate 100 ml volumetric flasks respectively. Both the drugs were dissolved in 50 ml of mobile phase with shaking and then volume was made up to the mark with mobile phase to get 100μg/ml and 300μg/ml of standard stock solution of ILA and DOM respectively. Several aliquots of standard solutions of ILA and DOM were taken in different 10 ml volumetric flasks and diluted up to the mark with mobile phase to get five different concentrations 5, 7.5, 10, 12.5, 15μg/ml and 15, 22.5, 30, 37.5,45μg/ml  of ILA and DOM respectively.


Preparation of sample solution:

Sample solution containing both the drugs was prepared by dissolving capsule powder into mobile phase. Twenty ILA and DOM capsules were weighed separately. Their average weights were determined. Powder of capsules equivalent to 10 mg of ILA and 30 mg of DOM were weighed and taken in a 100 ml volumetric flask, dissolved in mobile phase and sonicated for about 10 min then filtered through filter paper. An aliquot (1 ml) of filtered solution was further diluted in 10 ml volumetric flask with mobile phase to make the final concentration of working sample solution (10μg/ml ILA and 30μg/ml DOM).


Development and validation of HPLC method: Present study was conducted to obtain a new, affordable, cost-effective and convenient method for HPLC determination of ILA and DOM in capsule dosage form. The method was validated for the parameters like system suitability, selectivity, linearity, accuracy, and precision.


System suitability:

System suitability study of the method was carried out and various chromatographic parameters such as retention time, peak area tailing factor, theoretical plates of the column and resolution between the peaks were determined and the method was evaluated by analyzing these parameters.



Selectivity test determines the effect of excipients on the assay result. To determine the selectivity of the method, standard sample of ILA and DOM were injected first then commercial product solution were run in the instrument one after another.



Linearity of the method was determined by constructing calibration curves. Standard solution of ILA and DOM of different concentration levels 5-15μg/ml and 15- 45μg/ml of ILA and DOM respectively. The peak areas of the chromatograms were plotted against the concentrations to obtain the calibration curves and correlation coefficients.



The accuracy of the method was tested based on % Recovery by the assay of known and added amount of analyte. The recovery experiments were carried out in triplicate by spiking previously analyzed samples of the capsules (ILA 5μg/ml and DOM 15μg/ml) with three different concentrations of standards (ILA 4, 5,6μg/ml and DOM 12,15, 18μg/ml)



Intra-day precision was determined by performing three repeated analysis of the three standard solutions 5, 10 and 15μg/ml and 15, 30, 45μg/ml of ILA and DOM, respectively on the same day. On the other hand inter-day precision of the method was assessed by carrying out the analysis of standard solutions 5, 10 and 15μg/ml and 15, 30, 45μg/ml of ILA and DOM, respectively on three different days in the same laboratory. The relative standard deviation (% RSD) was calculated.


Limit of detection (LOD) and Limit of quantification (LOQ):

The LOD is defined as the smallest level of analyte that gives a measurable response. Limit of quantification LOQ is defined as the lowest concentration at which the precision expressed by relative standard deviation (RSD) is less than 2% and accuracy expressed by relative difference in the measured and true value is also less than 2%.


The LOD and LOQ for both ILA and DOM were determined according to ICH guideline Q2B1. The LOD and LOQ were estimated from the standard calibration curve. The residual standard deviation of regression line or standard deviation of y intercepts of regression lines used to calculate LOD and LOQ.


Here, LOD=3.3* D/S and LOQ=10*D/S.

Where, D is the standard deviation of y intercept of regression line and S is the slope of calibration curves.



Results of system suitability study of the standard solution showed uniform retention time, theoretical plate count, tailing factor and resolution for both the drugs which indicate a good system for analysis (Table I).


TABLE I-Result of system suitability tests of ILA and DOM.




Retention time (min)



Tailing factor



Resolution factor


Theoretical plate




The analytical characteristics of the proposed method derived from the calibration curves are as shown (Table II).

TABLE II-Analytical characteristics of the proposed method.




Linear Range (μg/ml)



Mean of Slope



S.D. of Intercept



Limit of Detection (μg/ml)



Limit of Quantification (μg/ml)



Regression coefficient (r2)




Chromatograms explain that retention time for standard sample and commercial product of ILA and DOM are same. This proves that, excipients have no effect on the analytical method. On the other hand, blank peak did not overlap drug peak. So the method is highly selective (Figure 3 and 4).


A linear relationship between peak areas versus concentrations was observed from 5-15μg/ml and 15-45μg/ml for ILA and DOM respectively. Correlation coefficient was 0.999 for both the drugs which prove that the method is linear (Figures 5 and 6).



Figure 3-Chromatogram of standard Ilaprazole and Domperidone



Figure 4-Chromatogram of sample and standard solution of Ilaprazole and Domperidone


Figure 5-Calibration curve of Ilaprazole


Figure 6-Calibration curve of Domperidone


Results of accuracy study were obtained by recovery test. Spiked amount of both the drug were compared against the recovery amount. % Recovery was 99.80-100.09 % for ILA and 99.40-99.77 % for DOM. All the results indicate that the method is highly accurate (Table III).





TABLE III-Accuracy (% Recovery) results of ILA and DOM.

Level of % recovery

Amount present


Amount of standard drug  added (μg/ml)

Total amount recovered (μg/ml) (n=3)

% Recovery






































Intra-day variability was done from 9.00 am to 6.00 pm on the same day. % RSD of peak areas was calculated for various run. The method is highly precise as % RSD of peak area was less than 2 % in all tests (Tables IV and V).


TABLE IV-Intraday and inter day precision result of ILA.

Conc. μg/ml

Intra-day Peak Area Mean ± Std. Deviation (n=3)


Inter-day Peak Area Mean ± Std. Deviation (n=3)



481.48466 ± 2.78579


479.0683 ± 5.6606



969.8600 ± 10.0702


967.1427 ± 12.17039



1454.32133 ± 13.3960


1452.6520 ± 12.3806



TABLE V-Intraday and inter day precision result of DOM.

Conc. μg/ml

Intra-day Peak Area Mean ± Std. Deviation (n=3)


Inter-day Peak Area Mean ± Std. Deviation (n=3)



915.3236 ± 10.2098


914.2053 ± 9.4772



1849.7546 ± 18.5217


1849.2300 ± 13.9903



2777.0893 ± 19.1634


2765.9240 ± 15.4274


The result of assay of was obtained and found to be in range 98%-102%. (Table VI).


Table VI- Amount of ILA and DOM in LUPILA-D Capsule by Proposed HPLC Method.

Amount present in (mg/cap)

Amount obtained in (mg/cap) (n=5)

Label Claim %















The proposed high-performance liquid chromatographic method has been evaluated for the accuracy, precision and linearity. The measured signals were shown to be precise, accurate and linear over the concentration range tested (5-15μg/ml of ILA and 15-45μg/ml of DOM) with a correlation coefficient of 0.999. In this method, there was no interference from matrix sources. Moreover, the lower solvent consumption along with the short analytical run time of 10 min leads to an environmentally friendly chromatographic procedure that allows the analysis of a large number of samples in a short period of time. Therefore, this HPLC method can be used as a routine sample analysis.



The author is thankful to Lupin Ltd. and Torrent Pharmaceutical for providing gift sample and Sardar Patel College of Pharmacy for providing the laboratory facilities and other requirements.



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Received on 16.05.2017          Accepted on 15.09.2017

© Asian Pharma Press All Right Reserved

Asian J. Pharm. Res. 2018; 8(1): 01-05.

DOI: 10.5958/2231-5691.2018.00001.1