Microsomal metabolism of calycosin, formononetin and drug-drug interactions by dynamic microdialysis sampling and HPLC-DAD-MS analysis.
A dynamic microdialysis sampling method with liquid chromatography-diode array detection and time-of-flight mass spectrometry (LC-DAD-TOF/MS) analysis was developed to investigate rat microsomal metabolisms of calycosin and formononetin, and their drug-drug interactions. Two hydroxylated metabolites from calycosin, and three hydroxylated or 4'-O-demethylated derivatives from formononetin were detected and identified after co-incubation with microsomes. Calibration curves offered linear ranges of two orders of magnitude with r(2)>0.999 for calycosin, formononetin and daidzein. The quantitative LC method provides a range of 0.028-0.034microg/mL for limits of detection, overall precision less than 5% and accuracy less than 3% by RSD. Besides, calycosin and formononetin were found to produce the depressive effect on the CYP450 enzyme reaction, and inhibit phase I enzyme reaction of each other when they are concurrent. Dynamic microdialysis sampling with LC-DAD-TOF/MS analysis developed in this work is a powerful tool for in vitro metabolism studies of drugs and metabolic interactions.
Wen
Xiao-Dong
XD
China Pharmaceutical University, Ministry of Education, Nanjing, China.
Qi
Lian-Wen
LW
Li
Bin
B
Li
Ping
P
Yi
Ling
L
Wang
Ya-Qiong
YQ
Liu
E-Hu
EH
Yang
Xiao-Lin
XL
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004347:Drug Interactions; D007529:Isoflavones / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D017551:Microdialysis; D008861:Microsomes / Q000378:metabolism*; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D007529:Isoflavones; C121707:7,3'-dihydroxy-4'-methoxyisoflavone; C007768:formononetin