Bioanalytical method to detect midazolam in plasma.

Midazolam (MDZ) is the first choice in palliative sedation, and commonly used in sleep induction in anesthesia, with rapid onset of action. However, monitoring of the level of sedation in patients is not accurate. We developed and validated a bioanalytical method to detect MDZ in plasma using high-performance liquid chromatography (HPLC) coupled to a photodiode array detector (PDA) for future monitoring of sedation. MDZ was extracted by solid-phase extraction (SPE). Analyses were performed on a C18 column, using 0.05% triethylamine and acetonitrile as mobile phase, analyzing at 220 nm. Recovery was evaluated by comparing extracted and nonextracted solutions. Precision, accuracy, linearity, limits of detection (LD) and quantification (LQ), specificity and selectivity were determined. The mean recovery obtained by SPE was 101.03%. The method was linear in the range 1.0-50.0 μg/mL. The LD and LQ were, respectively, 0.43 and 1.43 μg/mL. The specificity of the MDZ peak was adequate. The method was able to detect MDZ among other drugs. Plasma anticoagulants showed no interference with the drug detection. The bioanalytical method using HPLC-PDA and SPE was successfully validated and showed linearity, precision, accuracy, specificity and high sensitivity for detection of MDZ in human plasma.