International Journal of Innovative Pharmaceutical Sciences and Research

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1 International Journal of Innovative Pharmaceutical Sciences and Research SIMULTANEOUS ESTIMATION OF ATENOLOL AND CHLORTHALIDONE IN BULK AND COMBINED PHARMACEUTICAL DOSAGE FORM BY UV SPECTROSCOPY 1 Gurram Bhagath Kumar Goud*, 2 Sandhya, 3 Vasudha Bakshi Department of pharmaceutical Analysis and Quality Assurance, School of pharmacy, Anurag group of institutions, Venkatapur, Ranga reddy district, Telangana, INDIA Abstract A simple, accurate, precise and economical procedure for simultaneous estimation of Atenolol and Chlorthalidone in combined tablet dosage form has been developed utilizing concept of simultaneous equation method. The method is based upon determination of Atenolol and Chlorthalidone at 211nm and 221nm in methanol. Different analytical parameters such as linearity, precision, accuracy and ruggedness were determined according to ICH guidelines Atenolol and Chlorthalidone at their respective wavelength shows linearity in the concentration range of 5-40 mcg/ml and 1-10mcg/ml.respectively.The method was validated statistically. The results of analysis formulation given as percentage of label claim were found to be 99.55% and % for Atenolol and Chlorthalidone respectively. Therefore, the proposed method can be used for the routine analysis of both drugs simultaneously in quality control laboratories. Keywords: Atenolol, Chlorthalidone, simultaneous estimation, methanol. Corresponding Author Gurram Bhagath Kumar Goud Department of pharmaceutical Analysis and Quality Assurance School of pharmacy, Anurag group of institutions, Venkatapur, Ranga reddy district, Telangana, INDIA gurram.bhagathgoud52@gmail.com Mobile: Available online: November Issue 2749

2 INTRODUCTION Atenolol chemically, 2-[4-{(2RS)-2-hydroxy-3-[(1-methylethyl) amino] propoxy} phenyl] acetamide (Fig.1) is a selective β1 receptor antagonist, a drug belonging to the group of beta blockers, a class of drugs used primarily in cardiovascular diseases. The chemical works by slowing down the heart and reducing its workload. Atenolol does not pass through the blood-brain barrier thus avoiding various central nervous system side effects.[1] Atenolol is primarily used for hypertension, anginapectoris &myocardial infarction. I mainly act by inhibition of renin release and angiotensin-ii (AT-II) and aldosterone production. Chlorthalidone chemically 2-chloro-5-(1- hydroxyl-3-oxo-2, 3-dihydro-1H-isoindol-1-yl) benzene-1-sulfonamide (Fig.2) is widely used in antihypertensive pharmaceutical preparations, reduces active sodium reabsorption and peripheral vascular resistance. Chlortalidone is a diuretic drug used to treat hypertension. It is described as a thiazide diuretic. Compared with other medications of the thiazide class, chlorthalidone has the longest duration of action but a similar diuretic effect at maximal therapeutic doses. It is often used in the management of hypertension and edema. Literature survey reveals that there are several analytical methods for the estimation of atenolol and chlorthalidone individually or in combination with other drugs. Although the combination use of atenolol & chlorthalidone is continuously increasing, there is no UV method for the determination of these drugs in combined dosage form. Hence the aim of my study is to develop and validate a simple, precise, accurate, selective UV method for the estimation of atenolol and chlorthalidone in combined dosage form [1-12]. Fig.1: Structure of ATENOLOL Fig.2: Structure of CHLORTHALIDONE MATERIALS AND METHODS INSTRUMENT SPECIFICATIONS: UV Spectrophotometer, Shimadzu, model CHEMICALS AND REAGENTS: Methanol obtained from local market, manufactured Pure Atenolol and Chlorthalidone were obtained as gift sample from Startech chemical laboratories; Hyderabad.The tablet dosage form TENOCLOR-25( claim:25mg Atenolol and 12.5mg Chlorthalidone) was procured from local market. Available online: November Issue 2750

3 PREPARATION OF STANDARD STOCK SOLUTIONS The 10 mg of standard Atenolol and chlorthalidone were weighed accurately and transferred into 100 ml volumetric flasks seperately. They were dissolved in methanol and diluted up to the mark by using the solvent methanol to obtain a final concentration of 100μg/ml. The resulting solutions were used as working standard solutions. The aliquot portion of stock solutions of Atenolol and Chlorthalidone were diluted approximately with methanol to obtain concentration of 10 mcg/ml of each drugs.these solutions were scanned in the range of nm in 1cm cell against blank.from the overlain spectra the wavelength selected for the estimation are 211 nm and 221nm foratenolol and Chlorthalidone respectively. PREPARATION OF SAMPLE SOLUTIONS For analysis of commercial formulations 20 tablets (TENOCLOR-25) each cantaining 25mg of Atenolol and 12.5mg of Chlorthalidone were accurately weighed and the average weight was calculated and finely powdered.a quantity equivalent to 20mg dissolved in methanol in 10ml volumetric flask and pippet out 0.1ml in to another 10ml volumetric flask and adjust up to the mark this results 20mcg/ml solution which contains 20mcg/ml Atenolol and 5mcg/mlChlorthalidone sonicated to dissolve it.it is mixed well and filtered through 0.45µm filter to get the final concentration. This solution was scanned over the range of nm,using two sampling wavelengths 211nm and 221nm and determined the concentration of these drugs in tablet formulation using the formula. C ATN = A 2 ay 1 -A 1 ay 2 /ax 2 ay 1 -ax 1 ay 2 C CLT = A 1 ay 2 - A 2 ax 1 /ax 2 ay 1 -ax 1 ay 2 A 1 A 2 = absorbance of formulation at 211nm. = absorbance of formulation at 221nm. ax 1 = absorptivity ofatenolol at 211 nm. ax 2 = absorptivity ofatenolol at 221 nm. ay 1 = absorptivity of Chlorthalidone at 211nm. ay 2 = absorptivity of Chlorthalidone at 221 nm. C ATN = concentration of Atenolol C CLT = concentration of Chlorthalidone Available online: November Issue 2751

4 VALIDATION OF THE METHOD 1) LINEARITY: Standard stock solution of Atenolol and Chlorthalidone: 10 mg of Chlorthalidone and Atenolol were accurately weighed and transferred into a clean and dry 100 ml volumetric flasks, dissolved with sufficient volume of methanol seperately.the volume was then made up to 100 ml with methanol to obtain the concentration of 100 μg/ml. Working standard solution: The aliquot portion of the stock solution was diluted approximately with methanol to get a concentration of 1-10 mcg/ml of Chlorthalidone and 5-40mcg/ml of Atenolol. Serial dilutions of concentration range were prepared from the working standard solution. These dilutions were scanned at the detection wavelength of 211nm and 221nm using methanol as blank. The regression equation, Y- intercept and correlation coefficient were calculated. The linearity was thus determined and the concentration range was selected. 2) LIMIT OF DETECTION AND LIMIT OF QUANTITATION: The LOD and LOQ were separately determined based on calibration curve. The residual standard deviation of a regression line or the standard deviation of y- intercepts of regression lines were used to calculate the LOD and LOQ. I. Formula for LOD (μg/ml); LOD = 3.3 x SD / S Where, SD = The standard deviation of the response S = The slope of the calibration curve (mean) II. Formula for LOQ (μg/ml); LOQ = 10 x SD / S Where, SD = The standard deviation of the response S = The slope of the calibration curve (mean) 3) ACCURACY: The accuracy of the method was ascertained by carrying out recovery studies using standard method. The recovery studies are performed to determine if there was any positive or negative interference from excipients present in the formulation. The percentage recovery results revealed that the values were near to 100%, which indicates that the proposed method is accurate as the Available online: November Issue 2752

5 results are within the official limits. It also reveals that the commonly used excipients and additives in the formulation were not interfering with the proposed method. Table 1: Detection &Quantification limits in UV. Limit Wavelength(nm) Atenolol Chlorthalidone LOD 211nm nm LOQ 211nm nm ) PRECISION: Precision was found by carrying out the analysis of the standard drugs at two different concentrations in the linearity range of drugs for three times on the same day. Each concentration was applied in duplicate and % RSD was calculated (Table 2). Table 2: Precision Studies Drug Concentration Intra-day precision(n=3) Inter-day precision(n=3) (mcg/ml) %RSD %RSD Atenolol Chlorthalidone RESULTS AND DISCUSSION Atenolol and Chlothalidone showed maximum absorbance in Methanol at 211 and 221 nm.the proposed method for simultaneous estimation of both the drugs was validated as per the ICH guidelines The linearity was observed in the concentration range of 5-40 mcg/ml for Atenolol and 1-10mcg/ml for Chlorthalidone with regression co-efficient of 0.998,0.999(at 211nm & 221nm respectively) and & 0.998(at 211nm &221nm respetively).amount of drugs estimated by the proposed method was in good agreement with the label claim.the accuracy of the method was assessed by recovery experiments. Recovery was close to 100% for both the drugs Table 3: Calibration data of Atenolol Concetration (mcg/ml) 211 nm 221 nm Available online: November Issue 2753

6 Table 4: calibration data of Chlorthalidone Concetration (mcg/ml) 211 nm 221 nm Drug Table 5: Results of analysis of tablet formulation (TENOCLOR) Amount labled mg/tablet Amount estimatedmg/tablet % Lable clam %RSD Atenolol Chlorthalidone Table 6: Results of recovery studies Drug Recovery % Amount added Amount recovered (mcg/ml) (mcg/ml) % Recovery Atenolol Chlorthalidone Fig. 3: Overlain spectra of Atenolol and Chlorthalidone in methanol Fig.4: Formulation spectra Available online: November Issue 2754

7 Fig 5: Calibration curve of ATENOLOL at 211 nm Fig 6: Calibration curve of ATENOLOL at 221 nm Fig. 7: Calibration curve of CHLORTHALIDONE at 211 nm Fig. 8: Calibration curve of CHLORTHALIDONE at 221 nm CONCLUSION The present study comprises a UV spectroscopic method of analysis for the simultaneous estimation of Chlorthalidone and Atenolol in tablet dosage form.from the study of validation parameters,it was observed that the method is specific,accurate,precise,reproducible.the proposed method could be applied to routine analysis in quality control laboratories. REFERENCES Agon, P., Goethals, p., Van Haver, D. and Kaufman, J.M.: J. Pharm. Pharmacol., 43(8): (1991) Wikipedia.org/wiki/Chlorthalidone.\ 5. Permender Rathee., Sushila Rathee., Shyama Thakur. And Vikash Kumar.: International Journal of Chem. Tech. Research CODEN (USA).,2: (2010). 6. Vetushi, C. and Ragno, G.: International Journal of Pharmaceutics., 65: (1990). Available online: November Issue 2755

8 7. Salvador, C.S. and Luz, E.: J. Chromatog. Sci., 690: (1997). 8. El-Gindy., Alaa Sallam., Shehab. and Abdel Salam.: Jounal of Separation Sciences., 31(4): (2008). 9. Bilal, Y.: FABAD J. Pharm. Sci., 33: (2008). 10. De croo, F., Van Den Bossche, W.and De Moerloose.: Journal of Chromatogaphy A., 325: (1985). 11. Kavitha, J. An Muralidharan, S.: Internal Journal of Chem. Tech Research CODEN (USA)., 2: (2010). 12. Pankaj Kumara., Shubhanjali Shukla., Ashok Laxman Rao, G. and Subudhi, B.B.: Der Pharma Chemica., 3(2): (2011). Available online: November Issue 2756

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