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This study aimed to develop a reverse phase high-performance liquid chromatographic (RP-HPLC) method for the determination of efavirenz in human plasma and to use it for determining the concentrations of efavirenz in Chinese AIDS patient. A simple mobile phase consisting of 0.01 mol/L NaH2PO4 solution and acetonitrile (38:62, V/V) was pumped at a flow rate of 1.0 mL/min through a reverse phase Diamonsil C18 column maintained at 30°C. Diazepam was used as an internal standard and monitored with efavirenz at 247 nm. The protein of 100 μL plasma sample was precipitated before 20 μL of the supernatant was directly injected into the column. The linear response over the concentration ranges 0.10-20.0 μg/mL was obtained and the linear regression equations was Y = 2.2873X ‒ 0.1449 (r = 0.9999). The intra-day and inter-day precisions (1.9-2.6%, 2.2-7.2%, respectively), the relative and absolute recovery (99.3-106.3%, 75.6-80.3%, respectively) met the international standards. Stability of plasma samples were evaluated for short-term (ambient temperature for 16 h) and long-term (-20°C for 30 days) storage conditions and were found to be stable. The mean plasma concentration of efavirenz of the 406 patients was 2.21 ± 1.95 μg/mL, 77.3% of which were within the therapeutic window (1-4 μg/mL), 15.1% were below the window, and 7.6% were over it. In conclusion, the method had advantages of convenience, rapidity, necessary accuracy and precision, high practicality and met the needs for therapeutic drug monitoring and the pharmacokinetic study of efavirenz, especially in underdeveloped countries. For Chinese AIDS patients, it was beneficial to use efavirenz under the guidance of therapeutic drug monitoring.

bioscience trends

A simple, rapid, economical, and practical method for the determination of efavirenz in plasma of Chinese AIDS patients by reverse phase high-performance liquid chromatography with ultraviolet detector