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A simple and rapid reversed phase high performance liquid chromatographic method for quantification of alprazolam and alpha hydroxyalprazolam in plasma


A simple and rapid reversed phase high performance liquid chromatographic method for quantification of alprazolam and alpha hydroxyalprazolam in plasma



Journal of Liquid Chromatography 14(19): 3531-3546



ISSN/ISBN: 0148-3919

A simple and rapid reversed-phase liquid chromatographic method for the determination of alprozolam and a-hydroxyalprozolam in plasma is described. Flunitrazepam was used as internal standard. Plasma samples were buffered with sodium borate and extracted with dichloromethane/n-pentane 4:6 v/v for 60 sec on a vortex apparatus. Extraction solvent was evaporated to dryness and extraction residues were reconstituted in the mobile phase. Samples were chromatographed of a 5 .mu.m Lichrospher RP-18 column (25 cm .times. 4 mm i.d.) using acetonitrile/water 40:60 v/v as the mobile phase. The column effluent was monitored at 230 nm. The lower limit of detection was 1ng/ml for alprazolam and a -hydroxyalprazolam while the lower limit of quantification was 2 ng/ml for both compounds. Peak height and plasma alprazolam or a-hydroxyalprazolam concentrations were linearly related from 2.5 to 40 ng/ml. No potential sources of interferences have been identified. The methodology was used to monitor plasma alprazolam concentrations in patients receiving two single doses of alprazolam ranging from 0.25 to 1.0 mg each. Plasma alprazolam concentrations ranged from 4 to 27 ng/ml at two hours post second dose.

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Accession: 006966073

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