Analysis of midazolam and metabolites in plasma by high-performance liquid chromatography: probe of CYP3A.

Abstract:

:Hydroxylation of midazolam (MDZ) is mediated almost exclusively by CYP3A isoforms. The authors describe a high-performance liquid chromatography assay involving MDZ, 1'-hydroxymidazolam, and 4-hydroxymidazolam in plasma. The compounds were eluted on an Ultrasphere ODS, 3-microm particle size, 7.5 cm x 4.6 mm reversed-phase column and monitored by ultraviolet absorbance at 254 nm. The composition of the mobile phase was 35.2% acetonitrile:4.8% methanol:60% buffer acetate (vol/vol/vol), 0.1 M, pH 4.7; the flow rate was 1 ml/minute. Calibration curves were linear (coefficients of correlation > 0.99) within the range of concentrations established (20 to 640 nM). Within- and between-day coefficients of variation were consistently better than 8%. The overall recovery was >90% and the lowest detectable concentration was 8 nM. This approach provides a simple, rapid, and sensitive assessment of MDZ and metabolites in plasma, with a very good accuracy and precision, which enables it as an in vivo marker of CYP3A activity in humans.

journal_name

Ther Drug Monit

authors

Carrillo JA,Ramos SI,Agundez JA,Martinez C,Benitez J

doi

10.1097/00007691-199806000-00013

subject

Has Abstract

pub_date

1998-06-01 00:00:00

pages

319-24

issue

3

eissn

0163-4356

issn

1536-3694

journal_volume

20

pub_type

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