Eurasian J. Anal. Chem.

5(2): 145-151, 2010

New RP-HPLC Method for The Determination of Olmesartan Medoxomil in Tablet Dosage Form
Raveendra B.Gandurib*, Ramprasad A.Lankaa, Srinivasu Pamidia, Jayachandra R.Peddareddigarib and Mustafa Mohammedb
a. b

Analytical R&D, Hetero Drugs LTD, Hyderabad, Andhara Pradesh, India. HITS College of Pharmacy, Bogaram (V), Hyderabad, Andhara Pradesh, India.

Received: 16 July 2009; Accepted: 23 December 2009 Abstract A simple rapid, sensitive, accurate, precise and reproducible high performance liquid chromatographic method has been developed to assay Olmesartan medoxomil in tablet dosage form. The HPLC analysis used a reversed phase Hypersil BDS C8 (250X4.6mm, 5m) column and a mobile phase constituted of buffer and acetonitrile (55:45 % v/v). The buffer is composed of 3 g of sodium perchlorate and 3 mL of tri ethyl amine in 1000 mL of water and the pH of the solution was adjusted to 3.0 with orthophosphoric acid. The wave length of the detection is 250 nm. The validation data showed that the assay is sensitive, specific and reproducible for the determination of olmesartan in the dosage form. The method is linear from 10 g mL-1 to 120 g mL-1. The accuracy of the method was found to be 99.54%. Mean inter and intraday assay relative standard deviation (RSD) were less than 1.0%. The proposed method provided an accurate and precise analysis of olmesartan in its pharmaceutical dosage form. Keywords: Olmesartan medoxomil; antihypertensive; reversed-phase; validation

1. Introduction Olmesartan is chemically (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl-4-(1-hydroxy1methylethyl)-2-propyl-1-[4-[2-(tetrazole-5-yl)phenyl]methylimidazole-5-carboxylate. As a selective and competitive, nonpeptide angiotensin II receptor antagonist, olmesartan blocks the vasoconstrictor and aldosterone-secreting effects of angiotensin II [1-4]. A thorough literature has revealed that several methods were reported for the estimation of olmesartan in biological fluids [5-8].The techniques used include HPLC with mass [5-7], flourimetric [8] detections. The use of LC hyphenated techniques for identification of degradation products in stressed tablets of olmesartan was published in [9].Shinde et al have reported the stability indicating LC method for the determination of olmesartan in bulk drug and in pharmaceutical dosage form [10].Lisiane Bajerski et al developed a stability indicating LC determination of olmesartan medoxomil in tablets [11].Determination of olmesartan medoxomil in tablets by UV-Vis spectrophotometry was published in [12]. Piyush Trivedi et al reported a stability-indicating assay method for estimation of olmesartan medoxomil and its metabolite [13].
*

Corresponding Author E-mail: [email protected] ISSN: 1306-3057,

Moment Publication 2010

Ganduri et. al. Patel CV et al described validated absorption factor spectrophotometric and Reversephase High Performance Liquid Chromatography methods for the determination of Ramipril and olmesartan medoxomil in pharmaceutical formulations [14]. The objective of the proposed study was to assay olmesartan in its dosage form. We report the development and validation of a simple HPLC assay with UV detection for the quantitative determination of olmesartan in tablet dosage form. 2. Experimental 2.1 Chemicals and reagents All the reagents were of analytical-reagent grade unless stated otherwise. Glassdistilled and de-ionized water (Nanopure, Barnsted, USA), HPLC-grade acetonitrile, Sodium per chlorate, diethyl amine and orthophosphoric acid (S.D. Fine Chem., Mumbai, India) were used. 2.2 Instrumentation The HPLC system was composed of 2695 water alliance system fitted with 2996 PDA detector with empower software. Analytical column used for this method is Hypersil BDS C8 (250 mm x 4.6 mm) 5m (Thermo Electron Corporation, Runcorn, UK). 2.3 Buffer preparation Buffer solution was prepared by dissolving 3 gm of sodium perchlorate and 3 mL of triethyl amine in sufficient quantity of water and made up the volume to 100 mL (pH adjusted to 3.0 with ortho-phosphoric acid). 2.4 Standard Preparation Olmesartan medoxomil reference substance was accurately weighed (40 mg) and dissolved in a 20 mL quantity of acetonitrile: water (50:50) in a 100 mL volumetric flask and dilute up to the mark and it was further diluted to generate a concentration of 40 g mL-1. 2.5 Sample Preparation Twenty tablets of olmesartan (40 mg of olmesartan) were separately weighed and grounded to fine powder. An amount equivalent to 80mg of olmesartan was transferred into a 100 mL volumetric flask and dissolved in 60 mL quantity of methanol: water (50:50) and made up volume to 100mL. Further dilutions were made to generate a concentration of 40 g mL-1. 2.6 Chromatographic conditions Before the mobile phase was delivered into the system, buffer and acetonitrile were filtered through 0.45 m, PVDF membrane filter and degassed using vacuum. The chromatographic conditions used for the analysis were given below.
Column Wavelength Injection volume Flow rate Column temperature Run time Hypersil BDS C8 (250 mm x 4.6 mm) 5m 250 nm 10 l 1.0 mL min-1 250C 15 min

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Eurasian J. Anal. Chem. 5(2): 145-151, 2010

2.6 Method validation Method validation was conducted according to published guidelines [15-16]. Assay performance was evaluated by intraday and inter day (two different days) precision and determined from replicate analysis of samples (40 g mL-1) in two analytical runs. Analysis of six different sample solutions was performed in the same day for intraday precision. Accuracy of the method was tested by adding a known amount of olmesartan standard (40, 80 and 120 g mL-1) in three sample solutions. The precision and accuracy were expressed in terms of RSD from mean intra and inter day assays and recovery of the theoretical concentration. Robustness was tested by analysis of variations in analytical condition. Influence of mobile phase composition and different column brands were evaluated. The chromatographic parameters monitored were peak retention time, tailing factor and theoretical plate number. 3. Results and discussion Changes in the analytical procedure were tested. Different mobile phases with different proportions of organic modifier (acetonitrile) were tried. The pH value of the mobile phase was checked over a wide range (2.8-3.2). The pH of the aqueous phase was adjusted with orthophosphoricacid. Chromatographic run was evaluated using Hypersil BDS C8 column. After selecting the best conditions based on peak performance, the run time of the proposed assay was 15 min with isocratic elution. During injection of a standard and sample solution, the retention times were 6.954 and 6.966 min respectively (Fig.1).

Fig 1. (a) Chromatogram of the standard solution

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(b)

Fig 1. (b) Chromatogram of the sample solution 3.1 Interference studies 3.1.1 Excipient interference All the inactive ingredients namely hydroxyl propyl cellulose, lactose, magnesium stearate, microcrystalline cellulose and titanium dioxide were also tested in the proposed method. No peak was found at the retention time of olmesartan medoxomil when injected in to chromatographic system. 3.1.2 Impurity interference One of the degradant of olmesartan medoxomil, free olmesartan was injected into the chromatographic system at a level of 0.8 g mL-1. It had been found that it is not interfering with peak of interest. Forced degradation studies revealed that peak was pure in all the stress conditions. The results were shown in Table 1. Table 1. Forced Degradation studies
Stress condition Protected sample Water/Reflux-1.0 Hrs Acid degradation 0.01N HCl Reflux-1.0 Hrs Base degradation 0.01N NaoH Reflux-1.0 Peroxide degradation 1.5% H2O2 Reflux-1.0 Thermal degradation At 1050C-24 Hrs Photolytic degradation At 254nm-24 Hrs %Assay 99.1 97.3 97.9 96.8 97.0 91.2 91.0 %Degradation --1.8 1.2 2.3 2.1 8.0 8.2 Peak purity 0.99724 0.99902 0.99608 0.99955 0.99964 0.99586 0.99920

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3.1 Robustness Typical variations in analytical conditions were tested. Influence of flow rate, pH, mobile phase composition and filter variability were studied. The results were shown in the Table 2. Table 2. Robustness study of olmesartan
Robustness Parameter Variation in pH 2.8 3.0 (Optimized) 3.2 Mobile phase 50:50 Composition 55:45 (Optimized) 60:40 Variation in flow rate 0.8 mL min-1 1.0 (Optimized) 1.2 Filter variability Nylon Centrifuged PVDF Tailing factor 1.1 1.1 1.3 1.3 1.1 1.3 1.2 1.1 1.0 1.3 1.1 1.2 Theoretical plate number (n) 10176 13247 9754 9568 13247 11053 14560 13247 11221 15938 13247 14009 RSD (%) 0.08 0.05 0.10 0.11 0.05 0.12 0.08 0.05 0.05 0.11 0.05 0.08

3.2 Linearity The curve proved to be linear over a concentration range of 10-120 g mL-1 (Fig 2). Standard solution were prepared at seven concentrations (10, 20, 40, 60, 80, 100, 120 g mL-1) were injected in duplicate. Linear regression of concentration Vs peak area resulted in an average coefficient of determination (R2) 1.000. The percentage of Y-intercept is 0.150.
6000000

5000000

4000000 Area

3000000

2000000

1000000

0 0 20 40 60 80 100 120 140 Concentration (ug/m L)

Fig 2. Calibration curve of Olmesartan

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Ganduri et. al. 3.4 Precision The intra and inter day precision were estimated from duplicate injection of six sample solutions prepared at 40 g mL-1 of olmesartan analyzed on two different days. Mean and RSD were obtained from calculated olmesartan concentration (Table 3). The results indicate that the method is reproducible. Table 3. Intra and inter day precision for olmesartan
Sample No 1 2 3 4 5 6 Mean SD RSD (%) Intraday 99.6 101.2 98.5 100.2 99.1 99.8 99.7 0.93 0.93 Interday 99.2 100.7 99.1 99.5 98.8 100.2 99.6 0.88 0.89

3.3 Accuracy Accuracy was calculated as the percentage recovery of the known added amount of olmesartan reference substance in the sample solutions using three concentration levels covering the specified range (50,100,150 g mL-1) was added in the sample solutions (40 g mL-1). The accuracy of the method ranged from 98.7 to 100.3 %, indicating that this assay is reliable (Table 4) and meeting the acceptance criteria 98.0 to 102 %. Table 4. Accuracy of the analysis of olmesartan
Percent level 50 100 150 Added amount 40.26 80.11 119.82 Found amount 40.05 79.90 119.14 Recovery, % 99.46 99.73 99.43 RSD (%) 0.71 0.60 0.55

4. Conclusion The HPLC method developed and validated allows a simple and fast quantitative determination of olmesartan from its formulation. A mobile phase composed of solvent A and acetonitrile with a short run time (15 min) and isocratic elution used are advantageous and made the routine analysis easy. Among the significant advantages of this method are simplicity, selectivity, accuracy and precision ensuring that it is suitable for determining the content of olmesartan in tablet dosage form.

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Acknowledgements The authors are thankful to Hetero Drugs Limited, Hyderabad, India for providing reference standard and all facilities to complete this research work. References 1. 2. 3. Norwood D, Branch E, Smith B and Honeywell M (2002) Olmesartan medoxomil for hypertension: a clinical review, Drug forecast. P&T 27 (Suppl 12):611. Chrysant SG and Chrysant GS (2004) Expert Opin Pharmacother 37:657. doi:10.1517/ 14656566.5.3.657. Chrysant SG, Weber MA, Wang AC and Himman DJ (2004) Evaluation of antihypertensive therapy with the combination of olmesartan medoxomil and hydrochlorthiazide. Am J Hypertens 17:252. doi:10.1016/j.amjhyper.2003.11.003. Marshall TG, Lee RE and Marshall FE (2006) Common angiotensin receptor blockers may directly modulate the immune system via VDR,PPAR and CCR2b.Theor Biol Med Model 3:1.doi:10.1186/1742-4682-3-1. Vikas V (2008) LC-MS-MS determination of olmesartan in human plasma. Chromatographia 67(1):1 Chen SH (2008) HPLC-MS for determining olmesartan in human plasma. NanFang Vike Da Xue Xue Bao 28(60):1104. Liud (2007) Quantitative determination of olmesartan in human plasma and urine by LC coupled to tandem mass spectrometry. J Chromatogr B 856(1):190. Liu JF Wang (2006) Determination of olmesartan in human plasma by HPLC with fluorescence detection. J Pharm Anal 26(5):686 Murakami T, Konno H, Fukutsu, Onodera M, Kawasaki T and Kusu F (2008) Identification of a degradation product in stressed tablets of olmesartan medoxomil by the complementary use of HPLC hyphenated techniques. J Pharm Biomed Anal 47(3):553.

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