Application of microdialysis to study the in vitro metabolism of drugs in liver microsomes.
Current methods for studying in vitro drug metabolism involve add-incubate-separate-measure approach. Separation of the desired analytes requires removal of protein which is typically accomplished by precipitation and centrifugation and extraction of the analytes into an organic phase. The analysis scheme then becomes more complex resulting in a decrease in precision and an increase in assay time. Microdialysis sampling circumvents these problems by allowing researchers to sample the reaction mixture periodically and obtain the complete metabolic profile. In the present study, microdialysis sampling was used to investigate Phase I metabolism of salicylic acid, diazepam and ibuprofen in rat liver microsomes. The major metabolites of these drugs were profiled by LC. Michaelis-Menten enzyme kinetic parameters, Km and Vmax were obtained for the formation of diazepam metabolites by both microdialysis and conventional microsomal incubations and were in good agreement with the values reported in the literature. This study shows that microdialysis has considerable promise as a sampling technique for in vitro drug metabolism studies. By making minor modifications to the instruments, microdialysis can be applied to other in vitro systems such as isolated hepatocytes to study the Phase II metabolism or tissue slices to study drug distribution.
Gunaratna
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Bioanalytical Systems Inc., West Lafayette, IN 47906, USA.
Kissinger
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PT
D000818:Animals; D014151:Anti-Anxiety Agents / Q000378:metabolism*; D000889:Anti-Arrhythmia Agents / Q000378:metabolism; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003975:Diazepam / Q000378:metabolism*; D007052:Ibuprofen / Q000378:metabolism*; D007700:Kinetics; D017551:Microdialysis / Q000379:methods*; D008862:Microsomes, Liver / Q000378:metabolism*; D003708:Nordazepam / Q000032:analysis; D051381:Rats; D012459:Salicylates / Q000378:metabolism; D020156:Salicylic Acid; D013693:Temazepam / Q000032:analysis
D014151:Anti-Anxiety Agents; D000889:Anti-Arrhythmia Agents; D000894:Anti-Inflammatory Agents, Non-Steroidal; D012459:Salicylates; D003708:Nordazepam; D013693:Temazepam; D020156:Salicylic Acid; D003975:Diazepam; D007052:Ibuprofen