Development and Validation of HPTLC Method for Simultaneous Estimation of Amlodipine Besylate, Hydrochlorothiazide and Telmisartan In Their Combined Tablet Dosage Form

Abstract

Background: The combination of Amlodipine Besylaye, Hydrochlorthiazide and Telmisartan is prescribed for the treatment of hypertension. An Ultra Performance Liquid Chromatography (UPLC) method has been reported for simultaneous estimation of this combination. Objective: To develop and validate HPTLC Method for simultaneous estimation of Amlodipine Besylate, Hydrochlorothiazide and Telmisartan in their combined tablet dosage form. Materials and Method: The chromatographic separation was performed using aluminum plates pre-coated with silica gel 60F254 as stationary phase and chloroform: butan-1-ol: ammonia (6: 4: 0.1, v/v/v) as mobile phase. Spectro-densitometric scanning was performed at 254 nm. The developed method was validated according to ICH Q2R1 guideline. Results and Discussion: The linearity was established over a concentration range of 200-1000 ng/band, 500-2500 ng/band and 1600-8000 ng/band with correlation coefficient r 2 = 0.9952, 0.9992 and 0.9979 for Amlodipine besylate, Hydrochlorothiazide and Telmisartan, respectively. The R f values of Amlodipine besylate, Hydrochlorothiazide and Telmisartan were found to be 0.27 ± 0.02, 0.43 ± 0.02 and 0.14 ± 0.02 respectively. Recovery of drug was achieved in the range of 99.43–101.57%, 100.22–101.54% and 100.12–100.44% for Amlodipine besylate, Hydrochlorothiazide and Telmisartan, respectively by developed method. Limit of detection and limit of quantitation was found to be 8.6, 58.0 and 186.9 ng/band and 26.1, 175.8 and 566.4 ng/band for Amlodipine besylate, Hydrochlorothiazide and Telmisartan, respectively. Application: The developed HPTLC method was applied for simultaneous estimation of three drugs in their combined tablet dosage forms and results were found to be in good agreement with the labeled claim. Conclusion: The developed HPTLC method was found to be accurate, precise, specific and sensitive. It can be applied for routine analysis (assay) of tablets containing combination of Amlodipine besylate, Hydrochlorothiazide and Telmisartan. Key words: Amlodipine besylate (AML), Hydrochlorothiazide (HCTZ), Telmisartan (TLM), High Performance Thin Layer Chromatography (HPTLC), Simultaneous estimation.