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  • Title: Simultaneous determination of flumazenil, midazolam and metabolites in human biological fluids by liquid chromatography.
    Author: Chan K, Jones RD.
    Journal: J Chromatogr; 1993 Sep 08; 619(1):154-60. PubMed ID: 8245156.
    Abstract:
    A simple procedure for the simultaneous determination of flumazenil, midazolam, 1-hydroxymethylmidazolam and 4-hydroxymidazolam in plasma or urine in surgical patients is described. The assay involves a preliminary extraction of the drugs, metabolites and internal standard (flurazepam) from biological fluid into an organic solvent mixture (dichloromethane-diethyl ether, 40:60 v/v). The extract was evaporated to dryness at 40 degrees C under a gentle stream of nitrogen. The residue was re-dissolved in distilled methanol (80 microliters) and a 30-microliters aliquot was injected via an automatic sampler into the liquid chromatograph and eluted with the mobile phase (32% acetonitrile in 0.004 M sodium hydrogenphosphate buffer containing 1 ml of triethylamine and adjusted to pH 7.2) at a flow-rate of 1.5 ml/min on a 30-microns C8 precolumn linked to a 4-microns Nova-pak C18 cartridge column (100 mm x 8 mm I.D.) at ambient temperature (25 degrees C). The eluate was detected at 220 nm.
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