BONAGIRI SREEDEVI Pharmaceutical Chemistry Teegalaramreddy C JNTUH Hyderabad, INDIA Abstract: A new, simple, precise, accurate and reproducible RP
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1 Volume No., Issue No.,October November, 9 9. VANKAYALAPATI RAJYA LAKSHMI Pharmaceutical Chemistry Teegalaramreddy College Pharmacy JNTUH, JNTUH Hyderabad, INDIA. BONAGIRI SREEDEVI Pharmaceutical Chemistry Teegalaramreddy C Colllegee Pharmacy JNTUH, JNTUH Hyderabad, INDIA INDIA. Abstract: A new, simple, precise, accurate and reproducible RP RP-HPLC HPLC method for Simultaneous estimation in bulk and pharmaceutical formulations. S Seeparration ion w as succe su essfully ully aach hievved d on a Hypersil C8 (mm (mm X.mm X µ Make: Waters) or equivalent in an isocratic mode utiliz tiliziing ng KHPO buffer (ph.): Methanol (:v/v) at a a flowrate owrate of.8.8 ml/min ml/min vallidat idated d and nd the t e rrespo spon nsee as found found too be linear inear in n th the drug co conceentra ration ion ra angee f µg/ g/ml to µg/ml /ml for and µg/ g/ml mlto to µg/ml /ml for Olmesartan Olmesartan. Th Thee va aluee the he coorrel relattion on coe oeffi ficien nt w as found und to to bee.9.9 and for Olmesartan.. The LOD and LOQ for Olmesartan were found to be.8.8,, 9.8, respectively respectively.. The LOD and LOQ for were found to be..7 7,, 9.7 respectively. This method found to be good percentage recovery for were found to be.. and.. respectively indicates that the proposed method is highly accurate. The specificity the method shows good correlation between retention times standard standard with the sample so, the method specifically determines the analyte in the sample without interference from excipients tablet dosage forms. The method extensively validated according to ICH guidelines for Linearity, Range, Accu Accurracy, acy, Precision, Precision, Specificity and Robustness. Keywords: RPRP-HPLC; HPLC;, Olmesartan I. INTRODUCTION torvastatin: torvastatin (Lipitor) is a member the drug class known as statins. It is used for lowering cholesterol. is a competitive inhibitor hydroxymethylglutaryl-coenzyme hydroxymethylglutaryl coenzyme A HMG CoA) reductase, the rate HMG-CoA) rate--determining determining (HMG enzyme in cholesterol biosynthesis via the mevalonate pathway. HMGHMG-CoA CoA reduct reductase ase catalyzes the conversion HMG-CoA HMG CoA to mevalonate. acts primarily in the liver. Decreased hepatic cholesterol levels increases hepatic uptake cholesterol and reduces plasma cholesterol levels. necessary precautions. Olmesartan is also contraindicated in diabetes mellitus patients taking aliskiren. Figure : Chemical structure Olmesartan II. MATERIALS AND METHOD A. Instrumentation Figure: Chemical structure Of Olmesartan: Olmesartan Olmesartan is an antihypertensive agent, which belongs to the class medications called angiotensin II receptor blockers. It is indicated for the treatment high blood pressure and is marketed under the name Olmetec Olmetec. The FDA label includes a black box warning black-box injury and death to the fetus, so women child childbearing age need to be warned and take the 7 7 The separation carried out on HPLC system with Waters 9 alliance with binary HPLC pump, Waters 8 PDA detector, Waters Empower stware with Hypersil C88,, (mm (mm. ; µm) column. B. Chemicals and Reagents a gift sample by Lara drugs pvt Ltd., Hyderabad. KHPO, Methanol HPLC grade purchased from E. Merck (India) Ltd., Mumbai. Ortho phosphoric acid AR grade obtained from S.D. Fine Chemical Chemicalss All rights Reserved. Page 9
2 Volume No., Issue No.,October November, 9. Mumbai and mille Q water.. C. HPLC Conditions The column efficiency, resolution and peak asymmetry were calculated for the standard solutions (Table I).The values obtained demonstrated the suitability the system for the analysis this drug combinations, system suitability parameters may fall within ± standard deviation range during routine performance the method. The mobile phase consisting Phosphate buffer ph.: Methanol degassed and pumped from the solvent reservoir in the ratio :v/v pumped into the column at a flow rate.8 ml/min. The column temperature C. The detection monitored at nm and the run time min. The volume injection loop µl prior to injection the drug solution the column equilibrated for at least min. with the mobile phase flowing through the system. TABLE I: SYSTEM SUITABILITY PARAMETERS D. Preparation standard solution Accurately weigh and transfer mg and mg Olmesartan into ml volumetric flask and add ml mobile phase and sonicate min (or) shake min and makeup the volume with mobile phase. Pipette out.ml standard stock standard stock into ml volumetric flask dilute to volume with mobile phase and inject into HPLC. E. Preparation Of Solution Accurately weighed.mg sample. Transfer the sample powder into ml volumetric flask add ml mobile phase, sonicate for mins. Then make up the volume with mobile phase and filter through the.µm filter paper. Transfer ml above solution ml volumetric flask and make up the volume with mobile phase. System Suitability Studies Parameter Olmesartan Retention time Tailing factor.. RSD... Specificity Specificity is the ability to assess unequivocally the analyte in the presence components which may expect to be present. Typically these might include impurities, degradants, matrix, etc.. Accuracy and precision The accuracy the method determined by recovery experiments. The recovery studies were carried out six times. The percentage recovery and standard deviation the percentage recovery were calculated. From the data obtained, added recoveries standard drugs were found to be accurate (Table-II&III). TABLE II: ACCURACY FOR ATORVASTATIN Figure : Standard chromatogram for Figure : Formulation chromatogram for F. Method validation 7 Accu racy level weight added found Reco very All rights Reserved. Page
3 Volume No., Issue No.,October November, TABLE III: ACCURACY FOR OLMESAAN Accura cy level Sampl e weigh added μg/m l foun Recove ry Mea n Figure : AccuracyChromatograms- Figure 7: AccuracyChromatograms The precision the method demonstrated by inter-day and intra-day variation studies. In the intraday studies, six repeated injections standard and sample solutions were made and the response factor drug peaks and percentage RSD were calculated. In the inter-day variation studies. six repeated injections standard and sample solutions were made for three consecutive days and response factor drugs peaks and percentage RSD were calculated. The chromatograms three different levels shown in Figure, &7. From the data obtained, the developed RP-HPLC method found to be precise (Table-IV). TABLE IV: PRECISION STUDIES (ATOR) (OLM) Assay Assay (OLM ) (ATOR ) Figure : AccuracyChromatograms All rights Reserved. Page
4 Volume No., Issue No.,October November, 9.. Linearity and range The linearity the method determined at five concentration levels. The calibration curve constructed by plotting response factor against concentration drugs. The slope and intercept value for calibration curve Y=(R=.9) for and Y=9(R=) for Olmesartan. The results shows an excellent correlation exists between areas and concentration drugs within concentration range indicated above. The overlay chromatograms linearity for shows in figure and the results for calibration curves are given in figure 8 & 9. TABLE V: ROBUSTNESS FOR ATORVASTATIN USP Tailing Temp Temp-.. Flow-. Flow Table VI: ROBUSTNESS FOR OLMESAAN USP Tailing Temp Temp-..9 Flow Flow LOD&LOQ Limit quantification and detection were predicted by plotting linearity curve for different nominal concentrations and Olmesartan. RSD (σ) method predicted using following formulas (a) and (b). Precision established at this predicted level applied; the LOQ and LOD values were Figure 8: Linearity Curve for (a) LOQ = σ / S (b) LOD =. σ / S Where σ = residual standard deviation response S = slope the calibration curve AU OLMESAAN -.7 ATORVASTATIN -. Figure 9: Linearity Curve for Olmesartan.. Minutes Figure : Overlay chromatograms Linearity for.. Figure : Chromatograms for LOQ Robustness Robustness the method determined by making slight changes in the chromatographic conditions. It observed that there were no marked changes in the chromatograms which demonstrated that the RP-HPLC method developed, are robust (Table-V&VI). Figure : Chromatograms for LOD All rights Reserved. Page
5 Volume No., Issue No.,October November, 9. TABLE VII: LOD and LOQ for and Olmesartan LOD OLM.8 79 LOQ OLM.8 77 LOD ATOR LOQ ATOR.8 9 III. RESULTS AND DISCUSSION System suitability results were given by table and system suitability parameters are retention time, resolution, tailing and plate count were shown uniformity and RSD less than. So we can say system is suitable for analysis method specificity concluded by figure: and figure: those figures are standard chromatogram and other one is formulation they were not observed placebo and excipients peaks interference with standard and analytic peak so it proves method is selective. The result given in table I say that the method precision passed for both studies. The method accuracy evaluated by recovery studies. recovery founded & as per ICH 97- and also percentage RSD very low so method is accurate shown in table II&III. Linearity calibration curve given below fig: 8&9 and plot the graph three different concentrations versus areas to construct the linear regression equation and to calculate the value correlation co-efficient. Linear correlation found to be Y= (R=.9) for, Y=9 (R=) for Olmesartan. Method robustness results were given by table V&VI, LOQ and LOD Results were given by table VII. IV. ACKNOWLEDGEMENT I am thankful to department Pharmaceutical Analysis and Quality Assurance Teegalaramreddy College Pharmacy, Jawaharlal Nehru Technological University, Hyderabad, for providing instruments and analytical support. 7 REFERENCES [] B.Revanth reddy, G. Aravind, simultaneous estimation olmesartan and atorvastatin in bulk and formulation by using uv spectroscopy and RP-HPLC, Indian journal research in pharmacy and biotechnology, ISSN: 7, vol. (), pp. -, March. [] D.Nagavalli, Venkata suresh babu aluri, Development uv spectrophotometric method for the simultaneous estimation olmesartan medoxomil and atorvastatin calcium in tablet by simultaneous equation and first order derivative method, Journal pharmacy research, vol, no (). [] N.Nikita patel, R.Parag patel, A.Falguni tandel, S.Charmy kothari, A.Shailesh shah, Ratio derivative spectrophotometric method for simultaneous estimation olmesartan medoxomil and atorvastatin calcium in their combined tablet dosage form, International journal pharmacy and pharmaceutical sciences, ISSN- 97-9, vol.,. [] Kumanan Raghunathan, Jitendra M Reddy, and Manasa R. Stability indicating RPHPLC method development and validation Olmesartan medoxomil, Asian J Phar Biol Res., Issue:, vol.,. [] Chimalakonda Kameswara Rao, Kakumani Kishore Kumar,Maddala VijayaLaxmi, Polisetty Srinivasulu, Gutta Madhusudhan, Khagga Mukkanti, and Koduri Sai Venkata Srinivas, Development and Validation Stability Indicating LC Method for Olmesartan Medoxomil, American Journal Analytical Chemistry, vol., pp.,. [] M. Lakshmi Surekha, G.Kumara Swamy and D. Vinay Kumar, Development and Validation RP-HPLC Method for the Estimation in Bulk and Tablet Dosage Form, International Journal Pharma Sciences, ISSN: -8, vol., No.,pp.9-9,. [7] Şerife Hande Temir, Bedia Koçyiğit Kaymakçıoğlu,A validated HPLC method for analysis atorvastatin calcium in tablet dosage forms, MUSBED, vol.(), pp.77,. CONCLUSION The proposed HPLC method found to be simple, precise, accurate and sensitive for the simultaneous estimation and Olmesartan pharmaceutical dosage forms. Hence, this method can easily and conveniently adopt for rountine quality control analysis and Olmesartan pure and its pharmaceutical dosage forms. V. All rights Reserved. Page
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