Development of High-Performance Liquid Chromatographic Method for Determination of Brotizolam in Human Plasma. A Clinical Pharmacokinetic Study

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Abstract A high-performance liquid chromatographic method using an ultraviolet detector (HPLC-UV) was developed for the determination of brotizolam in human plasma. Brotizolam and triazolam, which was used as an internal standard (I.S.) in plasma, were detected by ultraviolet absorbance at 240 nm. Brotizolam in plasma was extracted by a rapid and simple procedure based on C18 bonded-phase extraction, and C8 reversed-phase HPLC separation. Determination of brotizolam was possible in the concentration range of 1.0–50.0 ng/mL. The detection limits of determination for brotizolam were 0.5 ng/mL. The mean recoveries of brotizolam added to plasma were more than 94.5% with a coefficient of variation of brotizolam of less than 7.3%. The method is applicable to pharmacokinetic study of brotizolam in plasma of healthy volunteers.

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