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SHORT COMMUNICATION
Year : 2012  |  Volume : 1  |  Issue : 2  |  Page : 129-132

A RP-HPLC method for simultaneous estimation of ondansetron and ranitidine in pharmaceutical formulation


1 Department of Pharmaceutical Analysis, J. S. S. College of Pharmacy, (Off Campus College of JSS University, Mysore), Karnataka, India
2 Vice-Chancellor, J. S. S. University, Mysore, Karnataka, India

Date of Web Publication27-Sep-2012

Correspondence Address:
S N Meyyanathan
Department of Pharmaceutical Analysis, J. S. S. College of Pharmacy, (Off Campus College of JSS University, Mysore), Ootacamund-643 001, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2278-344X.101723

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  Abstract 

A simple, selective, rapid, precise and economical reverse phase HPLC method has been developed for the simultaneous estimation of ondansetron and ranitidine from pharmaceutical dosage forms. The method was carried out using a Phenomenex column C 18 (250 mm × 4.6 mm, i.d 5 μ) with a mobile phase consisting of 50 mM potassium dihydrogen orthophosphate: acetonitrile (pH 6, ratio 60:40 v/v) at a flow rate of 0.8 ml/min. Detection was carried out at 222 nm. Pantoprazole was used as an internal standard. The retention time of ondansetron, ranitidine, and pantoprazole were found to be 6.4, 3.0 and 11.0 min, respectively. The developed method was validated in terms of its accuracy, precision, linearity, limit of detection, limit of quantitation, and solution stability. The proposed method can be used for the estimation of these drugs in a combined dosage form.

Keywords: Ondansetron, ranitidine, RP-HPLC method


How to cite this article:
Meyyanathan S N, Venkatesh D N, Krishnaveni N, Babu B, Jeyaprakash M R, Raja RB, Hemnath E, Suresh B. A RP-HPLC method for simultaneous estimation of ondansetron and ranitidine in pharmaceutical formulation. Int J Health Allied Sci 2012;1:129-32

How to cite this URL:
Meyyanathan S N, Venkatesh D N, Krishnaveni N, Babu B, Jeyaprakash M R, Raja RB, Hemnath E, Suresh B. A RP-HPLC method for simultaneous estimation of ondansetron and ranitidine in pharmaceutical formulation. Int J Health Allied Sci [serial online] 2012 [cited 2019 Sep 16];1:129-32. Available from: http://www.ijhas.in/text.asp?2012/1/2/129/101723

Ondansetron [1] is chemically known as 9-methyl-3-[(2-methyl-1H-imidazol-1-yl)methyl]-1, 2, 3, 9-tetrahydrocarbazol-4-one, and used as an antiemetic to treat nausea and vomiting following chemotherapy. Ranitidine [1] is chemically known as (E)-N-(2-((5-(dimethylaminomethyl)furan-2-yl)methylthio)ethyl)-N¢-methyl-2-nitroethene-1,1-diamine and used for the treatment of peptic ulcer. Many methods have been described in the literature for the determination of ranitidine and ondansetron individually and their combination with other drugs. [2],[3] However, a few analytical methods such as UV and HPLC have been reported for the simultaneous estimation of these drugs in a combined dosage form. [4],[5],[6] A fixed dose combination containing ondansetron (4 mg) and ranitidine (300 mg) is presently available in the market as a tablet dosage form. The aim of this work was to develop a rapid RP-HPLC method with an ultraviolet detection for the simultaneous estimation of ondansetron and ranitidine in pharmaceutical dosage forms. The present RP-HPLC method was validated following the ICH guidelines. [7],[8],[9]

Acetonitrile (HPLC grade) was procured from E. Merck (India) Ltd., Mumbai. Disodium hydrogen orthophosphate and orthophosphoric acid (AR grade) were procured from Qualigens Fine Chemicals, Mumbai. Water HPLC grade was obtained from a Milli-QRO water purification system. Reference standards of ranitidine and ondansetron are procured from Nectas Pharmaceuticals Ltd, Mumbai, and pantoprazole was procured from Kopran Pharmaceuticals Ltd., Mumbai, India. Chromatographic separation was performed on a Waters HPLC system with the following configurations: 1515 solvent delivery system (pump), Rheodyne 7725i injector with 100 μl loop, 2487 Dual wavelength absorbance detector. Breeze data station was applied for data collecting and processing, and Phenomenex column C 18 (250 mm × 4.6 mm, id 5 μ) was used for separation.

The mobile phase consisting a mixture of acetonitrile and 50 mM phosphate buffer (pH 6.0 adjusted with orthophosphoric acid) (60:40 v/v) was filtered through a 0.2 μm membrane filter and degassed. Standard stock solutions of 1 mg ml-1 of ondansetron, ranitidine, and pantoprazole were prepared separately using a mixture of water and acetonitrile in the ratio 1:1 v/v. From the standard stock solution, mixed standard solution was prepared to contain 12 μg ml-1 of ondansetron, 60 g ml-1 of ranitidine, and 100 μg ml-1 of pantoprazole as an internal standard. The mobile phase was delivered at a flow rate of 0.8 ml ml-1 with detection at 222 nm [Figure 1]. The injection volume was 50 ml, and analysis was performed at ambient temperature.
Figure 1: Overlay spectra of ranitidine, ondansetron, and pantoprazole (IS)

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Twenty tablets, each containing 4 mg of ondansetron and 300 mg of ranitidine (CANDOTAC-O), were weighed and finely powdered. A quantity of powder equivalent to label claim was weighed and transferred to a sintered glass crucible. To this 10 ml of 1 mg ml-1 solution of pantoprazole was added, and the drugs were extracted with three quantities, each consisting of 20 ml of mixture of acetonitrile and water (1:1 v/v). The combined extracts were made up to 100 ml using the same mixture of the mobile phase. The solution was further diluted to get a concentration of 12 μg ml-1 of ondansetron, 60 μg ml-1 of ranitidine (theoretical value), and 100 μg ml-1 of pantoprazole as an internal standard and used for the estimation. With the optimized chromatographic conditions, a steady baseline was recorded. The mixed standard solution was injected, and a chromatogram was recorded. The retention times of ondansetron, ranitidine, and pantoprazole were found to be 6.4, 3.0 and 11.0 min, respectively. The chromatographic parameters were calculated and are shown in [Table 1]. The similar procedure was followed for the sample solution obtained from the formulation. The response factor (peak area ratio of standard peak area and internal standard peak area) of the standard solution, and sample solution were calculated. The concentration of the drugs was calculated using the following formula:
Table 1: Chromatographic parameters

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A typical chromatogram of the sample solution is illustrated in [Figure 2]. The detection was carried out at 222 nm. The peak area ratios of standard and sample solutions were calculated. The assay procedure was repeated six times and the mean peak area ratio and mean weight of standard drugs were calculated. The percentage of individual drugs found in formulation, mean, standard were calculated and are presented in [Table 2]. The results of analysis shows that the amount of drugs was in good agreement with the label claim of the formulation.
Figure 2: Typical chromatogram of sample solution

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Table 2: Results of analysis of formulation and recovery studies

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The accuracy of the method was determined by recovery experiments. The recovery studies were carried out six times, and the percentage recovery and standard deviation of percentage recovery were calculated and are presented in [Table 2]. From the data obtained, added recoveries of standard drugs were found to be accurate. The intraday precision RSD and interday precision RSD values obtained were 0.70-1.82% and 0.85-1.74%, respectively. The linearity of the method was determined at six concentration levels ranging from 2 to 12 μg ml-1 for ondansetron and 10-60 μg ml-1 for ranitidine, respectively. The calibration curve was constructed by plotting the response factor against the concentration of drugs. The slope and intercept value for the calibration curve were y = 0.014x + 0.0016 (r2 = 0.9992) for ondansetron and y = 0.01x + 0.0071 (r2 = 0.9993) for ranitidine, respectively. These results indicated that there was an excellent correlation exists between the response factor and the concentration of drugs within the concentration range indicated above.

The limit of detection (LOD) and limit of quantification (LOQ) of the developed method were determined by injecting progressively low concentrations of standard solutions using the developed RP-HPLC method. The LOD is the smallest concentration of the analyte that gives a measurable response (a signal-to-noise ratio of 3). The LOD for ondansetron and ranitidine were found to be 12 ng ml-1 and 62 ng ml-1 , respectively. The LOQ is the smallest concentration of the analyte, which gives a response that can be accurately quantified (a signal-to-noise ratio of 10). The LOQ was 37 ng ml-1 and 187 ng ml-1 for ondansetron and ranitidine, respectively .

The ruggedness of the method was determined by carrying out the experiment on different instruments like Shimadzu HPLC (LC-10AT) and Waters HPLC by different operators using different columns of similar type like Hypersil C 18 , Phenomenex LUNA C 18 and Hichrom C 18 . Robustness of the method was determined by making a slight change in the chromatographic conditions. It was observed that no marked changes in the chromatograms demonstrated that the HPLC method developed are rugged and robust. In order to demonstrate the stability of both standard and sample solutions during analysis, both solutions were analyzed over a period of 5 h at room temperature. The results show that for both solutions, the retention time and peak area of ondansetron and ranitidine were remained almost unchanged (% RSD less than 2.0) and no significant degradation within the time period was observed. This indicated that both solutions were stable for at least 5 h, which was sufficient enough to complete the whole analytical process.

Limit of detection (LOD) and limit of quantification (LOQ) demonstrated the suitability of system for analysis of this drug combinations, system suitability parameters may fall within ±3 % standard deviation range during routine performance of the method.

The proposed RP-HPLC method for the simultaneous estimation of ondansetron and ranitidine in combined dosage forms was found to be accurate, precise, linear, rugged, robust, simple, and rapid. Hence, the present RP-HPLC method is suitable for the quality control of raw materials, formulations, and dissolution studies.

 
  References Top

1.The Merck Index. 11th ed. Vol. 1225. New Jersey, USA: John Wiley and Sons; 1989. p. 1454.  Back to cited text no. 1
    
2.Venkateshwaran TG, King DT, Stewart JT. HPLC determination of a metoclopramide and ondensetron mixture in 0.9% sodium chloride injection. J Liquid Chrom Related Tech 1993;133:291-5.  Back to cited text no. 2
    
3.Magdalene XK, Stella K, Michael T. A novel gradient HPLC method for simultaneous determination of ranitidine, methyl paraben and propyl paraben in oral liquid pharmaceutical formulation. J Pharm Biomed Anal 2005;38:763-7.  Back to cited text no. 3
    
4.Chin Ho, Huang H-M, Hsu S-Y, Shaw C-Y. Simultaneous High Performance Liquid Chromatographic analysis for famotidine, ranitidine HCl, cimetidine and nizatidine in commercial products. Drug Dev Ind Pharm 1999;25:379-85.  Back to cited text no. 4
    
5.Pillai S, Singhvi I. Spectrometric simultaneous estimation of ranitidine hydrochloride and ondansetron hydrochloride from tablet formulation. Indian J Pharm Sci 2007;69:601-4.  Back to cited text no. 5
  Medknow Journal  
6.Patel NM, Fursule RA, Shirkhedkar AA, Talele GS. Simultaneous estimation of ranitidine hydrochloride and ondensetron hydrochloride by reverse phase high performance liquid chromatography. Asian J Chem 2006;18:2691-4.  Back to cited text no. 6
    
7.Abdel Hamid ME, Sharma D. Simultaneous quantification of doxorubicin, lorazepam, metoclopramide, ondansetron and ranitidine in mixtures by liquid chromatographyTandem mass spectrometry. J Liquid Chrom Related Tech 2004;27:641-60.  Back to cited text no. 7
    
8.ICH, Q2A Text on validation of analytical procedures, International Conference on Harmonization 1994.  Back to cited text no. 8
    
9.ICH, Q3B Validation of analytical procedures: Methodology, International Conference on Harmonization, 1996.  Back to cited text no. 9
    


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2]


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