Abstract

SIMULTANEOUS QUANTIFICATION OF EMTRICITABINE AND TENOFOVIR DISOPROXIL FUMARATE IN SPIKED HUMAN PLASMA BY HPLC

Sunitha P. G.* and Kaliappan.Ilango

ABSTRACT

A rapid, precise, specific and accurate reverse phase high performance liquid chromatographic method has been developed and subjected to validation for determination of Emtricitabine(EMB) and Tenofovir disoproxil fumarate(TEN) in spiked human plasma. The chromatographic separation was achieved on Phenomenex Luna C18 (25cm x 4.6 mm, particle size 5μm) column with a mixture of acetonitrile:triethylamine (pH adjusted with glacial acetic acid to 3.5) in the ratio of 50:50(v/v) as mobile phase. UV detection was performed at 265 nm. The validation parameters such as for linearity, accuracy, precision, specificity and sensitivity were determined as per ICH guidelines. The proposed method was successfully applied for the quantitative estimation of dosage form available in market. The retention time of 2.1minutes and 5.8 minutes were obtained for EMB and TEN respectively. The calibration curve was linear over the concentration range of 0.5-2.5μg mL-1 for EMB and 1.5-7.5 μg mL-1 for TEN. The results of recovery studies indicate that the proposed method is applicable for the simultaneous estimation of EMB and TEN in pharmaceutical dosage form.

Keywords: Emtricitabine, Tenofovir, RP-HPLC, Plasma spiking, Dosage form, Validation.