ABSTRACT:
A simple, fast, reproducible, reliable and sensitive UPLC method was developed for the simultaneous estimation of anti-retroviral drugs in combination namely, Emtricitabine, Tenofovir disoproxil fumarate and Efavirenz. The chromatographic separation was carried out on a sub 2 micron particle size column Waters Acquity UPLC BEH C18, 100 x 2.1 mm, 1.7 µm to achieve more efficiency and better resolution in a gradient mode elution consisting of mobile phase A : Buffer (0.05% of Trifluro acetic acid in water) and mobile phase B: Methanol. Detection wavelength was set at 262 nm with a flow rate of 0.4ml/min. The retention times of Emtricitibine, Tenofovir disoproxil fumarate and Efavirenz was 0.6, 1.88 and 3.23 minute respectively. Under optimized conditions all three components were well separated from each other and also from main degradation impurities like Monoester impurity of Tenofovir, S-Oxide impurity of Emtricitabine and Amino alcohol and Quinoline impurity of Efavirenz. This was further supported by forced degradation studies. The % RSD was well within the limits and the correlation coefficient was 0.999.The developed method was validated as per ICH guidelines and can be used in routine quality testing of individual dosage forms and in combination.