Validation of a HPLC/MS method for simultaneous quantification of clonidine, morphine and its metabolites in human plasma.

ABSTRACT: A high-performance liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of morphine, morphine's major metabolites morphine-3-glucuronide and morphine-6-glucuronide, and clonidine, to support the pharmacokinetic analysis of an ongoing double-blinded randomized clinical trial that compares the use of morphine and clonidine in infants diagnosed with neonatal abstinence syndrome. Plasma samples were processed by solid-phase extraction and separated on an Inertsil ODS-3 (4 ?m) column using an 0.1% formic acid in water-0.1% formic acid in methanol gradient. Detection of the analytes was conducted in the positive multiple reaction monitoring mode. The range of quantitation was 1-1000?ng/mL for morphine, morphine-3-glucuronide and morphine-6-glucuronide, and 0.25-100?ng/mL for clonidine. Intra-day and inter-day accuracy and precision were??15% for all analytes across the quantitation range. Extraction recovery rates were??94% for morphine, ?90% for M3G, ?87% for M6G and???79% for clonidine. Matrix effect ranged from 85-94% for clonidine to 101-106% for M3G. The method fulfilled all predetermined acceptance criteria and required only 100??L of starting plasma volume. Furthermore, it was successfully applied to 30 clinical trial plasma samples.

SUBMITTER: Tang F 

PROVIDER: S-EPMC6559858 | biostudies-literature | 2019 Jul

REPOSITORIES: biostudies-literature