“…Therefore, our analytical approach, based on the extraction and instrumental detection of midazolam alone, cannot be applied when the detection and/or quantitation of the above metabolites are required, as in pharmacokinetic/pharmacodynamic studies 17,. 37 However, 4‐hydroxymidazolam is not detectable in human plasma following subtherapeutic midazolam doses,12 or detectable at sub‐ng/ml levels following therapeutic doses;25 in addition, the more important active metabolite 1‐hydroxymidazolam is always found in much lower plasma concentrations than midazolam 12,. 17 Thus, the described method may be usefully applied in cases of acute midazolam intoxication,38 within diagnostic procedures for brain death ascertainments,39–41 assays following fatal overdoses,6,, 24,, 42,, 43 or in cases of driving under the influence of drugs 44,.…”