Determination of midazolam and its major metabolite 1 '-hydroxymidazolam by high-performance liquid chromatography-electrospray mass spectrometry in plasma from children

Muchohi, S. N., Ward, Stephen ORCID: https://orcid.org/0000-0003-2331-3192, Preston, L., Newton, C. R. J. C., Edwards, Geoffrey and Kokwaro, G. O. (2005) 'Determination of midazolam and its major metabolite 1 '-hydroxymidazolam by high-performance liquid chromatography-electrospray mass spectrometry in plasma from children'. Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, Vol 821, Issue 1, pp. 1-7.

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Abstract

We have developed a sensitive, selective and reproducible reversed-phase high-performance liquid chromatography method coupled with electrospray ionization mass spectrometry (HPLC-ESI-MS) for the simultaneous quantification of midazolam (MDZ) and its major metabolite, 1'-hydroxymidazolam (1'-OHM) in a small volume (200 mu l) of human plasma. Midazolam, 1'-OHM and 1'-chlordiazepoxide (internal standard) were extracted from alkalinised (pH 9.5) spiked and clinical plasma samples using a single step liquid-liquid extraction with 1-chlorobutane. The chromatographic separation was performed on a reversed-phase HyPURITy (TM) Elite C18 (5 mu m particle size; 100 mm x 2.1 mm i.d.) analytical column using an acidic (pH 2.8) mobile phase (water-acetonitrile; 75:25% (v/v) containing formic acid (0.1%, v/v)) delivered at a flow-rate of 200 mu l/min. The mass spectrometer was operated in the positive ion mode at the protonated-molecular ions [M + 1](+) of parent drug and metabolite. Calibration curves in spiked plasma were linear (r(2) > 0.99) from 15 to 600 ng/ml (MDZ) and 5-200 ng/ml (1'-OHM). The limits of detection and quantification were 2 and 5 ng/ml, respectively, for both MDZ and V-OHM. The mean relative recoveries at 40 and 600 ng/ml (MDZ) were 79.4 +/- 3.1% (n = 6) and 84.2 +/- 4.7% (n = 8), respectively; for 1'-OHM! at 30 and 200 ng/ml the values were 89.9 +/- 7.2% (n = 6) and 86.9 +/- 5.6% (n 8), respectively. The intra-assay and inter-assay coefficients of variation (CVs) for MDZ were less than 8%, and for 1'-OHM were less than 13%. There was no interference from other commonly used antimalarials, antipyretic drugs and antibiotics. The method was successfully applied to a pharmacokinetic study of MDZ and 1'-OHM in children with severe malaria and convulsions following administration of MDZ either intravenously (i.v.) or intramuscularly (i.m.). (c) 2005 Elsevier B.V. All rights reserved.

Item Type:	Article
Uncontrolled Keywords:	midazolam 1 '-hydroxymidazolam pharmacokinetics in children alpha-hydroxy metabolite gas-chromatography human serum status epilepticus pediatric-patients seizures diazepam 1-hydroxymidazolam pharmacokinetics flumazenil
Subjects:	QV Pharmacology > QV 38 Drug action. QV Pharmacology > QV 4 General works QV Pharmacology > Drug Standardization. Pharmacognosy. Medicinal Plants > QV 771 Standardization and evaluation of drugs
Faculty: Department:	Groups (2002 - 2012) > Molecular & Biochemical Parasitology Group
Digital Object Identifer (DOI):	https://doi.org/10.1016/j.jchromb.2005.03.015
Depositing User:	Sarah Lewis-Newton
Date Deposited:	10 Nov 2011 17:30
Last Modified:	17 Jul 2020 10:58
URI:	https://archive.lstmed.ac.uk/id/eprint/1914

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