Validated RP-HPLC Method for Simultaneous Estimation of Atenolol, Hydrochlorothiazide and Losartan Potassium in Bulk and Pharmaceutical Dosage Form
A simple, specific, accurate, and precise reverse phase high performance liquid chromatographic (RP-HPLC) method was developed for the simultaneous estimation of Atenolol, Hydrochlorothiazide and Losartan potassium in bulk and pharmaceutical dosage form. The separation was carried out using Hypersil C18, 250 ×4.6 mm, 5µm column with mobile phase consisted mixture of Acetonitrile and Potassium dihydrogen ortho phosphate buffer in the ratio of 40:60 (V/V) delivered at a flow rate of 1.5 mL / min and effluents were monitored at 225 nm. The selected chromatographic conditions were effective separation of these drugs. And the retention times of Atenolol, Hydrochlorothiazide and Losartan potassium were found to be 1.46, 2.21 and 3.30 min respectively. The proposed method was found to be linear in the range of 50-150 μg/mL,12.5-37.50 μg/mL, and 50-150 μg/mL for Atenolol, Hydrochlorothiazide and Losartan potassium respectively. The recovery of drugs was found to be 97.56 %, 97.72 % and 98.06 % respectively. The proposed method was validated as per ICH guidelines and it was found to be accurate, precise and robust, and it was applied to the estimation of Atenolol, Hydrochlorothiazide and Losartan potassium in combined tablet dosage form and it can also be used for routine quality control analysis of these drugs in biological samples either alone or in combined pharmaceutical dosage forms.
2. Data Base of Atenolol, Compilation Prepared by Drug Bank. Available from: http://www.Drug bank.ca/drugs/DB00335.
3. Indian Pharmacopoeia, Vol 1. Government of India, Ministry of Health and Family Welfare, Controller of Publications, Delhi. 1996; 72–73.
4. British Pharmacopoeia, Vol 1. The stationary office, London. 2005; 179–181.
5. United States Pharmacopoeia, 28th edition, Vol 1. Rockville, MD: The United States Pharmacopeial Convention. 2005;193.
6. Data Base of Hydrochlorothiazide, Compilation Prepared by Drug Bank. Available from: http://www.Drug bank.ca/drugs/DB00999.
7. Goodman LS, Gilman A. The Pharmacological basis of Therapeutics, 11th edition Diuretics. 2006; 753.
8. Data Base of Losartan, Compilation Prepared by Drug Bank. Available from: http://www.Drugbank.ca/drugs/db00678.
9. United States Pharmacopoeia/National Formulary-24 United States Pharmacopoeial Convention, Inc; Rockville, MD,2006, 1280-1282.
10. Bhatia N M, Desai R B and Jadhav S. Simultaneous Estimation of Losartan Potassium and Hydrochlorothiazide from Tablets by First Order Derivative Spectroscopy. International Journal of Pharmacy and Pharmaceutical Sciences. 2013;5(1):464-466.
11. Kasture A V, Ramteke. Simultaneous UV-spectrophotometric method for the estimation of atenolol and amlodipine besylate in combined dosage form. Indian J Pharm Sci. 2006; 68(3):394-396.
12. Al-Ghannam S M. A simple spectrophotometric method for the determination of β-blockers in dosage forms. J Pharm Biomed Anal. 2006; 40(1):151-156.
13. Erk N. Application of first derivative UV-spectrophotometry and ratio derivatives spectrophotometry for simultaneous determination of
14. Candesartan cilexetil and Hydrochlorothiazide. Pharmazie. 2003; 58:796–800.
15. Charles J J, Brault S, Boyer C, Langlois M H, Cabrero L, Dubost J P. Simultaneous determination of Irbesartan and Hydrochlorothiazide in tablets by derivative spectrophotometry. Anal Lett.2003;36(11):2485–2495.
16. Prabhakar A H, Giridhar R. A rapid colorimetric method for the determination of losartan potassium in bulk and in synthetic mixture for solid dosage form. J Pharm Biomed Anal. 2002;27(6):861-866.
17. Cagigal E, Gonzalez L, Alonso R M, Jimenez R M. Experimental Design Methodologies to Optimise the Spectrofluorimetric Determination of Losartan and Valsartan in Human Urine, Talanta. 2001;54(6): 1121-1133.
18. Chiguru Vishnuvardhan, P. Radhakrishnan and, Sameer G. Navalgund, Krishnam Raju Atcha, N. Satheesh kumar. RP-HPLC Method for the Simultaneous Estimation of Eight Cardiovascular Drugs. Chromatographia. 2014; 77:265–275. DOI 10.1007/s10337-013-2598-0.
19. Savita S Yadav and Janhavi R Rao. RP-HPLC Method for Simultaneous Estimation of Losartan, Hydrochlorothiazide and Amlodipine in Tablet Dosage Form. Asian J Pharm Clin Res. 2014;7(1):137-140.
20. Ravisankar Panchumarthy, Devala Rao Garikapati. An improved Rapid HPLC Method for the Separation of Five Anti-Hypertensive Agents Using C18 Column: Application to Hydrochlorothiazide determination in bulk and Tablet dosage form. IOSR Journal of Pharmacy. 2013;3(5):7-19.
21. M.M. Eswarudu, T. Narendra Chary, Sunil J, M. Sushma. HPLC Method Development and Validation for Simultaneous Estimation of Irbesartan and Hydrochlorothiazide in Pharmaceutical Dosage Form. Asian J. Research Chem.2012;5(4):348-352.
22. Sharma Rajesh, Sunil Khanna, Mishra Ganesh Prasad. Development and validation of RP-HPLC method for simultaneous estimation of Losartan potassium and Atorvastatin calcium in pharmaceutical preparations. J of Phar Res.2012;5(1):398-400.
23. Ramesh D, Habibuddin MD, Dash RN, Humaira T. Simultaneous determination of Atorvastatin calcium and Losartan potassium in bulk and combined dosage forms by validated RPHPLC with UV detection. Am. J. Pharm Tech Res.2012; 3(1): 675-685.
24. Ganesh GS, Shalini NS, Sistla S. Stability indicating RP- HPLC method for the simultaneous estimation of Atorvastatin and Losartan in bulk and pharmaceutical formulation. J of Sci Res in Phar. 2012;1(3): 53-56.
25. Vijayalakshmi R, Kalyani P, Sandya P, Dhana Raju MD. RP-HPLC method for the simultaneous estimation of Atorvastatin and Losartan in pure and tablet formulations. Int J of Res Phar and Chem.2012;2(3):885-888.
26. R. A. Mhaske, S. Sahasrabudhe and A. A. Mhaske. RP-HPLC method for simultaneous determination of Irbesartan, Losartan, Hydro-Chlorothiazide and Chlorthalidone–application to commercially available drug products. International journal of pharmaceutical sciences and research.2012;3(4):1116-23.
27. Neela M. Bhatia, Sachin B. Gurav, Swapnil D. Jadhav &Manish S. Bhatia, RP-HPLC method for simultaneous estimation of Atorvastatin calcium, Losartan potassium, Atenolol, and Aspirin from tablet dosage form and plasma. Journal of Liquid Chromatography & Related Technologies. 2012;35(3): 428-443.
28. Md. Arif Hossen, Md. Ahsanul Haque, Irin Dewan, A. N. M. Hamidul Kabir, Md. Khalid Hossain and S. M. Ashraful Islam. RP-HPLC Method for the Simultaneous Estimation of Hydrochlorothiazide and Losartan Potassium in Tablet Dosage. Dhaka Univ. J. Pharm. Sci. 2011;10(1):35-42.
29. Krishna R. Gupta, Akshay R. Wadodkar and Sudhir G. Wadodkar. Validated Reverse Phase HPLC Method for Simultaneous Estimation of Atorvastatin and Atenolol in Tablets. Der Pharmacia Lettre. 2011;3(4):393-403.
30. Dubey R, Bhusari VK, Dhaneshwar LR. Validated RP-HPLC method for simultaneous quantitation of Losartan potassium and Metolazone in bulk drug and formulation. Sci Pharm.2011;79: 545–554.
31. Zaveri Maitreyi, Amit Khandhar. Development and Validation of a RP-HPLC for the Simultaneous estimation of Atenolol and Hydrochlorothiazide in Pharmaceutical Dosage Forms. International journal of advances in Pharma science.2010;1(2):167-171.
32. Kumar Naveen, Verma N, Singh Omveer, Joshi N, Kanwar G S; Estimation of Atenolol by Reverse Phase High Performance Liquid Chromatography. E J Chem. 2010;7(3):962-966.
33. Rao KS and Srinivas K. RP-HPLC method for the determination of Losartan potassium and Ramipril in combined dosage form. Indian J Pharm Sci.2010;72(1):108-111.
34. Abdussaleem.K, D. Boopathy, Perumal. Development and Validation of Losartan Potassium and Atenolol in combined dosage form by RP-HPLC. International Journal of Pharm Tech Research.2010;2(1):471-474.
35. Barman R K, Islam M A U, Ahmed M et al. Simultaneous high-performance liquid chromatographic determination of Atenolol and Amlodipine in pharmaceutical-dosage form, Pak J Pharm Sci. 2007; 20(4):274-279.
36. Sivakumar T, Venkatesan P, Manavalan R, Valliappan K. Development of a HPLC method for the simultaneous determination of losartan potassium and atenolol in tablets. Indian J Pharm Sci. 2007; 69(1):154-157.
37. Panchal HJ and Suhagia BN. Simultaneous analysis of Atorvastatin calcium and Losartan potassium in tablet dosage forms by RP-HPLC and HPTLC. Acta Chromatographica 2010; 22:173-187.
38. Patel N D, Captain A D, Parmar K E, Development and Validation of HPTLC Method for Simultaneous Determination of Atenolol and Losartan Potassium in Bulk and in Pharmaceutical Dosage Form, International Journal of Pharmacy and Pharmaceutical Sciences, 2013;5(2):325-331.
39. NJ Shah, BN Suhagia, RR Shah, NM Patel. Development and validation of a HPTLC method for the simultaneous estimation of Irbesartan and hydrochlorothiazide in tablet dosage form, Indian journal of Pharmaceutical science.2007;69(2):202-205.
40. Argekar A P, Powar S G. Simultaneous determination of atenolol and amlodipine in tablets by high-performance thin-layer chromatography. J Pharm Biomed Anal.2000; 21(6):1137-1142.
41. Santhi Neelima M, Mohan Gandhi B, Bhaskara Raju V, Srinivas Sumanth K, Srinivas, K, Mounika P, Jhansi Naga Lakshmi P. Development and validation of stability indicating reverse phase high performance liquid chromatography method for simultaneous estimation of atenolol, hydrochlorothiazide and losartan in bulk and pharmaceutical dosage form. Asian journal of pharmaceutical and clinical research.2016;9(2):118-124.
42. Savita S Yadav, Janhavi R. Rao. Simultaneous estimation of Losartan, Hydrochlorothiazide and Atenolol from solid dosage form by RP-HPLC. International Journal of Pharmacy and Pharmaceutical Sciences. 2014; 6(1):283-288.
43. Tengli AR and Gurupadayya BM. Method development and validation of tablet dosage form containing Losartan, Atenolol and Hydrochlorothiazide using internal standard by RP-HPLC. Journal of Chromatography Separation Techniques.2013;4(5):1-5. doi:10.4172/2157-7064.1000180.
44. Sharma R, Khanna S, Mishra G P, RP-HPLC Method for Simultaneous Estimation of Atenolol, Hydrochlorothiazide and Losartan in Tablet Dosage Form, Chem Sci Trans., 2013, 2(S1), S1-S6.
45. Parthiban C, Bhagavan Raju M, Sudhakar M. Simultaneous estimation and validation of Atenolol, Hydrochlorothiazide and Losartan in tablet dosage form by RP-HPLC method. International Journal of Pharmacy & Industrial Research.2011;1(4):325-329.
A. B. Thomas, U. B. Chavan, R. K. Nanda, L. P. Kothapalli, S. N. Jagdale, S. B. Dighe and A. D. Deshpande, Simultaneous RP-HPLC analysis of atenolol, hydrochlorothiazide, and losartan potassium in a tablet formulation. AK Journals Acta Chromatographica, 2010;22(2): 219-226.
46. Kavitha J, Muralidharan J. Development and Validation of New Method for Atenolol, Hydrochlorothiazide and Losartan potassium by RP-HPLC: Its Application to Routine Quality Control Analysis. International Journal of Chem.Tech Research, 2010;2(2):880-884.
47. Rao DD, Satyanarayana NV, Sait SS, Reddy YR, Mukkanti K. Simultaneous Determination of Losartan Potassium, Atenolol and Hydrochlorothiazide in Pharmaceutical Preparations by Stability-Indicating UPLC. Chromatographia.2009;70(3-4):647-651.
48. Sathe SR, Bari SB, Simultaneous Analysis of Losartan Potassium, Atenolol and Hydrochlorothiazide in Bulk and in Tablets by High-Performance Thin-layer Chromatography with UV Absorption Densitometry. Acta Chromatogr. 2007; 19: 270–278.
49. Lavanya G, Sunil M, Eswarudu MM, Eswaraiah MC, Harisudha K and Spandana BN. Analytical Method Validation: An Updated Review. Int J Pharm Sci Res. 2013, 4(4), 1280-1286.
50. ICH Harmonized Tripartite Guideline, Validation of analytical procedures: Text and methodology, Q2 (R1), International Conference on Harmonization, Geneva. 2005; 1-13.
51. ICH Harmonized Tripartite Guideline Stability Testing of New Drug Substances and Products Q1A (R2), International Conference on Harmonization, Geneva. 2003; 1-18.
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
AUTHORS WHO PUBLISH WITH THIS JOURNAL AGREE TO THE FOLLOWING TERMS:
Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial 4.0 Unported License. that allows others to share the work with an acknowledgment of the work's authorship and initial publication in this journal.
Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in this journal.
Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).