Abstract
A pharmacokinetic interaction study between efavirenz (EFV), a non-nucleoside reverse transcriptase inhibitor used in the treatment of HIV-1 infection, and an African traditional medicine, African potato in human subjects was undertaken. This necessitated the development and validation of a quantitative method for the analysis of EFV in plasma. A simple mobile phase consisting of 0.1M formic acid, acetonitrile and methanol (43:52:5) was pumped at a low flow rate of 0.3ml/min through a reverse phase Phenomenex® Luna C18 (2) (5μm, 150mm×2.0mm i.d.) column maintained at 40°C. Diclofenac sodium was used as an internal standard (IS) and EFV and IS were monitored at 247nm and 275nm, respectively. A simple and rapid sample preparation involved the addition of mobile phase to 100μl of plasma to precipitate plasma proteins followed by direct injection of 10μl of supernatant onto the column. The procedures were validated according to international standards with good reproducibility and linear response (r=0.9990). The intra- and inter-day accuracies were between 12.3 and 17.7% at the LLOQ and between −5.8 and 9.1% for the QC samples. The intra- and inter-day precision of EFV determinations were 5.1 or less and 7.2% RSD or less, respectively across the entire QC concentration range. Mean recovery based on high, medium and low quality control standards ranged between 92.7 and 94.1% with %RSD values better than 3%. Plasma samples were evaluated for short-term (ambient temperature for 6h) and long-term (−10±2°C for 60 days) storage conditions and were found to be stable. The method described is cost-effective and has the necessary accuracy and precision for the rapid quantitative determination of EFV in human plasma.
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