Automation of metabolic stability studies in microsomes, cytosol and plasma using a 215 Gilson liquid handler.
A 215 Gilson liquid handler was used to automate enzymatic incubations using microsomes, cytosol and plasma. The design of automated protocols are described. They were based on the use of 96 deep well plates and on HPLC-based methods for assaying the substrate. The assessment of those protocols was made with comparison between manual and automated incubations, reliability and reproducibility of automated incubations in microsomes and cytosol. Examples of the use of those programs in metabolic studies in drug research, i.e. metabolic screening in microsomes and plasma were shown. Even rapid processes (with disappearance half lives as low as 1 min) can be analysed. This work demonstrates how stability studies can be automated to save time, render experiments involving human biological media less hazardous and may be improve inter-laboratory reproducibility.
Linget
J M
JM
Laboratoire de recherche Glaxo Wellcome France, Les Ulis.
du Vignaud
P
P
D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D017076:Computer-Aided Design; D003600:Cytosol / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D008861:Microsomes / Q000378:metabolism*; D010078:Oxazines / Q000032:analysis; D010949:Plasma / Q000378:metabolism; D051381:Rats; D015203:Reproducibility of Results
D000097:Acetonitriles; D010078:Oxazines; C014180:resorufin; C032159:acetonitrile