Determination of midazolam and its metabolites in serum microsamples by high-performance liquid chromatography and its application to pharmacokinetics in rats.
A single-solvent extraction step high-performance liquid chromatographic method is described for quantitating midazolam and its two hydroxy metabolites in rat serum microsamples (50 microliters). The separation used a 2 mm I.D. reversed-phase Symmetry C18 column with an isocratic mobile phase consisting of methanol-acetonitrile-14.9 mM sodium acetate in water at pH 3.0 (10:23:67, v/v). The detection limit was 10 ng/ml for all the compounds using an ultraviolet detector operated at 230 nm. The method was used to study the pharmacokinetics of midazolam after an intravenous bolus dose (0.75 mg/kg).
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Department of Psychology, Rutgers University, New Brunswick, NJ 08903, USA.
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D000818:Animals; D014151:Anti-Anxiety Agents / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D002940:Circadian Rhythm; D006993:Hypnotics and Sedatives / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D008297:Male; D008874:Midazolam / Q000097:blood* / Q000378:metabolism* / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D014151:Anti-Anxiety Agents; D006993:Hypnotics and Sedatives; D008874:Midazolam