SIMPLE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC DETERMINATION OF THE PROTEASE INHIBITOR INDINAVIR IN HUMAN PLASMA

Indinavir is a member of a class of protease inhibitors that actively prevent the acquired immunodeficiency syndrome virion from maturing. A high-performance liquid chromatographic (HPLC) assay was developed an d validated for the determination of indinavir in human plasma. Indina vir and the internal standard were isolated from the plasma by ether e xtraction. The residue after evaporation of ether was reconstituted wi th buffer and injected onto a C-4 reversed-phase column eluted isocrat ically with a mobile phase consisting of 35:65 (v/v) of acetonitrile a nd buffer. A wavelength of 210 nm was found to he optimum for detectio n. The calibration range of this assay was from 10 to 5000 ng/ml and c oefficients of variation For the assay ranged from 4.6% to 11.0% for t hree different drug concentrations and the limit of quantitation was 1 0 ng/ml. During the validation, short-term stability of the drug in pl asma, stability during heat deactivation and on repeated freezing and thawing of plasma was evaluated. The overall recovery of indinavir by the ether extraction method was 91.4%. This HPLC assay was found to be a simple and reproducible method for monitoring indinavir levels in h uman plasma obtained during clinical trials of the drug. (C) 1998 Else vier Science B.V.