Analytical method development and validation of related substances by rp-hplc of emtricitabine and tenofovir disoproxil fumarate tablets

Jay Bhatt*, Ujashkumar A Shah, Hirak kumar V Joshi, J K Patel

Faculty of Pharmacy, Nootan Pharmacy College, Sankalchand Patel University, Visnagar, Gujarat, India

ABSTRACT

The developed method was a simple, accurate, precise, specific and robust method for the validation of Emtricitabine and Tenofovir isoproxil Fumarate Tablets by reverse phase high pressure liquid chromatography. For Emtricitabine and Tenofovir Disoproxil Fumarate Chromatography was performed on Agilent 1200 series, UV and PDA Detector, Waters X-bridge C18 (250 mm x 4.6 mm, 5 μm) by preparing Buffer solution: Dissolve 0.63 g of ammonium formate in 1000 mL of purified water and mix. Adjust to pH of 3.90 +0.05 with diluted formic acid. And used it as mobile phase A. Mobile Phase B: mixture of buffer solution and methanol in the ratio of (20 : 80) % v/v at a flow rate of 1.0 mL/min and at 254 nm wavelength. The retention times of Emtricitabine and Tenofovir Disoproxil Fumarate are approx. 29 min and 70 min. respectively. 5-Fluorocytosinc, Sulfoxide Impurity Isomer 1, Sulfoxide Impurity isomer 2, 5-Fluorouracil analogue, Tenofovir (PMPA) Impurity, Monoester Impurity and Dimer Impurity found linear over the range of LOQ - 150 % of target concentration. Method also found precise by spiking impurities at specification level. Accuracy was demonstrate at LOQ - 150 % level by preparing sample in triplicate for each level and found accurate. Hence, the method could be successfully used for the analysis Impurities in Emtricitabine and Tenofovir Disoproxil Fumarate Tablets.

Keywords:

Emtricitabine, Tenofovir Disoproxil Fumarate, HPLC, UV and PDA Detector, Related Substances