On‐line desorption of dried blood spots coupled to hydrophilic interaction/reversed‐phase LC/MS/MS system for the simultaneous analysis of drugs and their polar metabolites

Abstract

An assay for the simultaneous analysis of pharmaceutical compounds and their metabolites from micro-whole blood samples (i.e. 5 microL) was developed using an on-line dried blood spot (on-line DBS) device coupled with hydrophilic interaction/reversed-phase (HILIC/RP) LC/MS/MS. Filter paper is directly integrated to the LC device using a homemade inox desorption cell. Without any sample pretreatment, analytes are desorbed from the paper towards an automated system of valves linking a zwitterionic-HILIC column to an RP C18 column. In the same run, the polar fraction is separated by the zwitterionic-HILIC column while the non-polar fraction is eluted on the RP C18. Both fractions are detected by IT-MS operating in full scan mode for the survey scan and in product ion mode for the dependant scan using an ESI source. The procedure was evaluated by the simultaneous qualitative analysis of four probes and their relative phase I and II metabolites spiked in whole blood. In addition, the method was successfully applied to the in vivo monitoring of buprenorphine metabolism after the administration of an intraperitoneal injection of 30 mg/kg on adult female Wistar rat.

Serially coupling hydrophobic interaction and reversed-phase chromatography with simultaneous gradients provides greater coverage of the metabolome

“…RPLC/HILIC systems have been utilized in many research areas, including metabolomics (Di Palma et al 2011;Gilar et al 2005;Kalili and de Villiers 2010;Wang and Lehmann 2008;Lam et al 2010). Although the solvents used for RPLC and HILIC are the same, the need for diametrically opposed organic concentration at the start of each separation has led to the development of on-line RPLC/ HILIC valve switching systems Thomas et al 2010;Wang and Lehmann 2008). These methods, although successful, are not suited for routine use in most laboratories as dedicated interfaces are required for handling mobile phase incompatibilities.…”

Quantification of Glucuronide Metabolites in Biological Matrices by LC-MS/MS

“…In step three, the flow is directed only through the RP column, where the non-polars are separated and eluted to MS. This 2D orthogonal approach combines the benefits of both RP and HILIC methods and provides an excellent sensitivity (Thomas, Déglon et al 2010). …”

Analyse des cannabinoïdes par spectrométrie de masse en mode tandem

“…C'est le cas de Thomas et coll. [9] qui pré-sentent une méthode de désorption de spots de sang séchés recueillis sur du papier buvard suivie de l'analyse par LC-MS/MS. Dans ce cas, une goutte de sang capillaire (5 μL) est prélevée au bout du doigt et recueillie sur un papier buvard.…”