Simultaneous Estimation of Nebivolol and Hydrochlorothiazide in ...

Asian J. Research Chem. 1(2): Oct.-Dec. 2008,,ISSN 0974-4169www.ajrconline.orgRESEARCH ARTICLESimultaneous Estimation of Nebivolol and Hydrochlorothiazide in CombinedTablet Dosage Form by Multicomponent Mode of AnalysisPS Tarte*, SP Wate, PB Khedikar, BT Satpure and G PawnikarSharad Pawar College of Pharmacy, Wanadongari, Hingana Road, Nagpur-441110 (M.S.)*Corresponding Author E-mail: pawantarte@rediffmail.comABSTRACTA simple, accurate, precise and economical procedure for simultaneous estimation of Nebivolol hydrochloride andHydrochlorothiazide in two component tablet dosage form has been developed utilizing concept of standard addition. Themethod is based upon determination of Nebivolol at 281 nm and Hydrochlorothiazide at 272 nm, in aqueous methanol (20% v/v). Nebivolol hydrochloride and Hydrochlorothiazide at their respective λ max 281.0 nm and 272.0 nm shows linearityin the concentration range of 5-35 µg/ml and 10-70 µg/ml respectively. The method was validated statistically. Recoverystudy was performed to confirm the accuracy of the method.KEY WORDSNebivolol hydrochloride, Hydrochlorothiazide, simultaneous estimationINTRODUCTION:The combination of Nebivolol (NBV) andHydrochlorothiazide (HCTZ) is used in hypertension.Chemically Nebivolol and Hydrochlorothiazide areα,α’-[Iminobis(methylene)]bis[6-fluoro-3,4-dihydro-2H-1-benzpyran-2-methanol] and 6-Chloro-3,4-dihydro-7-sulfamoyl-2H-1,2,4-benzothiadiazine 1,1-dioxide respectively. Nebivolol is a 1:1 racemicmixture of the enantiomeric pair, SRRR (d) & RSSS(l). 1,2 The β 1-antagonist actions are attributed primarilyto the d-isomer & both d - & l-isomers furthercontribute to the pharmacological profile of Nebivololthrough vascular endothelial nitric oxide releasingcapabilities. 3,4 Hydrochlorothiazide inhibits theabsorption of sodium and chloride at the beginning ofdistal convoluted tubule. In this combination,Hydrochlorothiazide is official in IP, BP and USP andmany analytical methods are reported for it. Nebivololis not official in any pharmacopoeia but somespectrofluorimetric and HPLC methods are reported for5,6it. There is, however, no work reported oncombination of these two drugs. In the presentcommunication, we propose fast, simple and accuratespectrophotometric method for simultaneousestimation of both the drugs in tablet dosage form.Received on 25.11.2008 Modified on 30.11.2008Accepted on 20.12.2008 © AJRC All right reservedAsian J. Research Chem. 1(2): Oct.-Dec. 2008;Page 77-79MATERIAL AND METHOD:Instrument:1) UV/Visible spectrophotometer SHIMADZU UV-1700with 1cm UV matched quartz cells2) One Pan (K-14 super By K Roy) BalanceReagent:Double distilled water and methanol of analytical grade.Preparation of standard stock solution:Accurately weighed quantity (100 mg) of Nebivolol (NBV)and Hydrochlorothiazide (HCTZ) were dissolved separatelyin 20 ml methanol and volume was made up 100 ml withwater (1000 µg/ml). The aliquot portions of stock solutionsof NBV and HCTZ were diluted appropriately with aqueousmethanol (20%v/v) to obtain concentration 20 µg/ml ofNBV and 10 µg/ml of HCTZ. These solutions were scannedin the range of 200-400 nm in 1cm cell against blank. Fromthe overlain spectrum shown in Fig 1, the wavelengthselected for estimation are 284 nm and 272 nm. The sixmixed standard solutions with concentration of NBV andHCTZ in the ratio of 5:10, 10:20, 15:30, 20:40, 25:50, 30:60(µg/ml) were prepared in aqueous methanol (20% v/v).ALLTHE mixed standard solutions were scanned over the rangeof 200-400 nm, in multicomponent mode; using twosampling wavelength 281nm (λ max of NBV ) and 272 nm(λ max of HCTZ ) . The spectral data from these scans wereused to determine the concentration of drugs in tabletformulation.Procedure for Analysis of Tablet Formulation:Twenty tablets were weighed, average weight determinedand finally powdered. Appropriate quantity of powder fromeach tablet equivalent to 5 mg NBV and12.5 mg HCTZ wasaccurately weighed and following standard addition77

method, 7,8 20 mg of pure NBV was added to achieve2:1 ratio and shaken vigorously with methanol for 15minutes and filtered. Necessary dilutions are made withaqueous methanol to give final concentration 20 µg/mland 10 µg/ml of NBV and HCTZ respectively. Thesample was scanned over the range of 200-400 nm inthe multicomponent mode and the concentration ofeach component were obtained by the spectral data ofthe sample solutions with reference to that of the mixedstandard. Results of analysis of tablet formulation areshown in Table 1.Table 1: Results of Analysis of Tablet Formulation andStatistical DataSampleLabelclaim%label SD RSD SEmg/tablet claim*NBV 5 mg 99.98 0.3278 0.0032 0.1465BrandHCTZI99.95 0.3305 0.0033 0.147812.5 mgNBV 5 mg 99.99 0.3048 0.0030 0.1363BrandHCTZII99.97 0.3088 0.0030 0138012.5 mg*Results are mean of five replicatesAsian J. Research Chem. 1(2): Oct.-Dec. 2008,,Linearity and range study was done by preparingconcentration in the range of 80 -120% of the testconcentration and absorbance values were recorded at 281nm and at 272 nm. The plot of linearity and range is shownin Fig 2.Absorbance1.510.5Linearity and rangeHCTZy = 0.0085x + 0.008R 2 = 0.9986272 nm281 nmLi (272 )NBVy = 0.0061x + 0.0054R 2 = 0.998300 50 100 150% ConcentrationFigure 2: Plot of linearity and range study of Nebivolol andHydrochlorothiazideRESULTS AND DISCUSSION:The proposed method for simultaneous estimation of NBVand HCTZ dosage forms were found to be simple, accurate,economical and rapid. The method was validated as per theICH and USP guidelines. The values of SD or RSD arewithin the prescribed limit of 2 %, showing high precisionof method and recovery was close to 100% for both thedrugs. During the linearity study it was observed thatabsorbance values of NBV and HCTZ in the marketedformulation were linear in the range of 80% to 120% of testconcentration with R 2 close to one.Figure 1: Overlain spectra of Nebivolol andHydrochlorothiazide9, 10Validation of method:The proposed method was validated on the basis ofparameters namely accuracy, precision, specificity,ruggedness and linearity and range.The accuracy of the method was ascertained bycarrying out recovery studies using standard additionmethod. The recovery study was performed todetermine if there was any positive or negativeinterference from excipients present in the formulation.The precision of an analytical method is expressed asSD or RSD of a series of measurements. It wasascertained by replicate estimation of drug by theproposed method.Test for ruggedness was carried out by repeating theprocedure under different conditions, i.e., on differentdays, at different time and by different analysts.Table 2: Results of Recovery StudyQuantity of drug % Recovery of % Recovery ofadded (µg/ml) brand* I brand* IINBV HCTZ NBV HCTZ NBV HCTZ10 5 99.42 99.43 99.10 99.7520 10 99.85 99.86 98.97 99.5530 15 99.66 99.68 99.38 99.42*Results are mean of five replicatesTable 3: Results of Ruggedness StudyCondition% Label claim SD RSDNBV HCTZ NBV HCTZ NBV HCTZIntraday 100.01 99.98 0.25320.250.000.00253225Interday 99.82 99.79 0.12120.110.000.00128511Different0.160.0099.83 99.81 0.1626 0.0016analyst5016*Results are mean of five replicatesCONCLUSION:The present study comprises an Absorbance Ratio methodto determine NBV and HCTZ from tablet dosage form.From the study of validation parameters namely accuracy,precision (SD and RSD), ruggedness (interday, intraday anddifferent analyst), specificity, linearity and range, it wasobserved that the method is specific, accurate, precise78

eproducible and rugged. The proposed method couldbe applied to routine analysis in quality controllaboratories.ACKNOWLEDGEMENTS:The authors are thankful to Dr. K. P. Bhusari,Principal, Sharad Pawar College of Pharmacy, Nagpur,for his valuable guidance and advice. The authors aregrateful to M/s Hetero Drug Limited, Medak and M/sGolden Cross, Daman for providing gift sample ofNebivolol and Hydrochlorothiazide respectively.REFERENCES:1. Budavari S. The Merck index: An Encyclopedia ofChemicals, Drugs and Biologicals. 13 th ed. WhitehouseStation (NJ): Merck Research Lab, Division of Merck;2001.1152.2. Sweetman SC. Eds., In; Martindale, The Complete DrugReference, London; Pharmaceutical Press; Division ofthe Royal Pharmaceutical Society of Great Britain,2007. (Electronic version. Available on http://www.medicinecomplete.com)3. Budavari S. The Merck Index: An Encyclopedia ofChemicals, Drugs and Biologicals. 13 th ed. WhitehouseStation (NJ): Merck Research Lab. Division of Merck;2001. 854.4. http:/www.xbl.com5. Rajeshwari KR, Sankar GG and Rao AL. Developmentof spectroflurometric method for the estimation ofNebivolol in tablets and human serum. 57 th IndianPharmaceutical Congress, Scientific Abstracts, 2005,298.6. Rajeshwari KR, Sankar GG and Rao SJVLN. RP-HPLCmethod for the estimation of Nebivolol in bulk andpharmaceutical dosage form. 57 th Indian PharmaceuticalCongress, Scientific Abstracts, 2004, 85.7. Chandna S, Kasture AV and Yeole PG. Simultaneousspectrophotometric determination of, Piogalitazonehydrochloride and Glimepiride in tablets. Indian JPharm Sci. 2005; 67: 6278. Sahu R, Nagar P, Bhatacharya S and Jain D.Simultaneous spectrophotometric estimation ofFamotidine and Domperidone in combined tablet dosageform. Indian J Pharm Sci. 2006; 68:503.9. ICH Q2A; Guidelines on validation of analyticalprocedure; Definitions and terminology, FederalRegister, 1995, 60, 11260.10. ICH Q2B; Guidelines on validation of analyticalprocedure; Methadololgy, Federal Register, 1996, 60,27464.Asian J. Research Chem. 1(2): Oct.-Dec. 2008,,79