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Simultaneous Estimation of Tenofovir and Emtricitabine in Human Plasma Using HPLC after Protein Precipitation Extraction

Arti Soni and Seema Thakral

Tenofovir (TNF) and emtricitabine (FTC) are both reverse transcriptase inhibitors, often used in combination for anti-retroviral therapy. In the present study, a reverse phase high performance liquid chromatographic method was developed and validated for the simultaneous estimation of TNF and FTC in human plasma using stavudine as the internal standard. Protein precipitation extraction procedure utilizing perchloric acid was employed to extract the drugs from human plasma. Estimation of the drug contents was done by using a mixture of sodium dihydrogen orthophosphate buffer (pH 6.9) and methanol as the mobile phase and absorbance was read at 259 nm for TNF and 280 nm for FTC. Retention time was found to be 6.6 (± 0.1) min for TNF and 17.1 (± 0.2) min for FTC. The recovery, selectivity, linearity, precision as well as the accuracy of the method was evaluated from spiked human plasma samples during the course of validation. The result revealed that the analytical technique presented here demonstrates acceptable accuracy and precision, shorter and simpler sample preparation, and a reduced need for complicated equipment along with a tolerable analysis time.

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