Development and validation of UHPLC-MS/MS method for analysis of sedative drugs and their metabolites in blood plasma

Date

2016

Journal Title

Journal ISSN

Volume Title

Publisher

Tartu Ülikool

Abstract

In intensive care units, the precise administration of sedative drugs is crucial in order to avoid under- or over sedation – both of which can be very harmful. It is especially important in case of paediatric patients. A highly sensitive simultaneous UHPLC-MS/MS method was developed for the quantification of morphine, morphine-3-β-glucuronide, morphine-6-β-glucuronide, clonidine, midazolam and 1’-hydroximidazolam in human plasma samples. The lowest limit of quantification for all analytes was 50 pg/mL using only 100 μL of blood plasma. Analytes were separated chromatographically using C18 column with weak ion-pairing additive 1,1,1,3,3,3-hexafluoro-2-propanol with pH = 9 (adjusted using ammonium hydroxide) and methanol. Method was fully validated and method was using matrix matched calibration in the range of 0.05-250 ng/mL for all analytes. Within-day accuracy for all analytes remained 87-113 %, but within-day precision remained within 3-11 % for all analytes at all concentration levels over the calibration range. The method will be applied for real patients’ samples in the EU FP7 project CloSed – "Clonidine for Sedation of Paediatric Patients in the Intensive Care Unit”.

Description

Keywords

UHPLC-MS/MS, blood plasma, sedative drugs, metabolites, morphine, morphine-3-β-glucuronide

Citation