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.

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.

Genetic structure of the Lesser Gymnure (Genus Hylomys) in SE-Asia: evidence for two species

The Lesser Gymnure is a small galericine Insectivore living in the mainland forests of Southeast Asia, including the major islands of Sumatra, Java and Borneo. It is the only representative of the supposed monospecific genus Hylomys which is morphologically rather constant over its geographic range. Only marginal subspecific differentiation is currently recognized based in slight pelage colour variations. We report here the results of 35 gene loci revealed by protein electrophoresis on 23 specimens sampled over most of Southeast Asia. They were compared with outgroup, Erinaceaus europeaus, member of a distinct subfamily. The surveyed populations of Lesser Gymnures clearly group themselves into distinct taxa, one of which seems restricted to Sumatra, while the other occupies the whole geographic range. The genetic distance between these groups is two times greater than the divergence observed within groups: it is of the same order of magnitude as what is usually reported for congeneric mammal species, which supports their specific distinction. The lack of gene flow is also demonstrated by several diagnostic loci defining unambiguously each species. Both are only distantly related to the outgroup, a result which is consistant with their actual classification into two distinct subfamilies (Erinaceae and Galericinae). Concordant genetic and geographic subdivision of the widespread species further suggest that eustatic sea level changes during the Pleistocene produced predictable patterns in species differentiation.