✍ Scribed by P. Odou; H. Robert; M. Luyckx; C. Brunet; T. Dine; B. Gressier; M. Cazin; J. C. Cazin
No coin nor oath required. For personal study only.
A routine high-performance liquid chromatographic method for measuring midazolam in human serum has been developed. The sample preparation procedure consisted of simple tiquid-liquid extraction with dichloromethane, followed by evaporation under nitrogen. The mobile phase used was a mixture of acetonitrile at 0.02 M and sodium acetate at pH 3.0 (80:20, vlv) and a flow-rate of 1.2 mWmin. The separation was performed on two cyanopropyl columns (150 x 4.6 mm). The detection was by UV absorption at 240 nm. A linear range from 10 to 1000 ng/mL and a quantification limit of 7.4 ng/mL of serum was reached. The mean intra-assay and inter-assay reproducibility from serum sample spiked with 100 ng/mL were 4.1 and 4.7%, respectively. The recoveries from serum sample spiked with 50, 100, 500 nglmL were 85.46, 85.38 and 85.57%, respectively. This method was developed to allow pharmacokinetic study of midazolam in young patients in short surgical interventions.
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