Analytical Sciences


Abstract − Analytical Sciences, 34(9), 1011 (2018).

Sensitive and Comprehensive LC-MS/MS Analyses of Chiral Pharmaceuticals and Their Hepatic Metabolites Using Ovomucoid Column
Kenichiro TODOROKI,*1 Yuto KUDOH,*1 Misuzu NAKAMURA,*1 Yuki SHIMIZU,*2 Takamitsu SASAKI,*2 Hideyuki OTSUKI,*3 Keisuke WADA,*3 Jun Zhe MIN,*4 Hajime MIZUNO,*1 Kouichi YOSHINARI,*2 and Toshimasa TOYO’OKA*1
*1 Laboratory of Analytical and Bio-Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka 422-8526, Japan
*2 Laboratory of Molecular Toxicology, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga, Shizuoka 422-8526, Japan
*3 Shinwa Chemical Industries Ltd., 50-2 Kagekatsu-cho, Fushimi, Kyoto 612-8307, Japan
*4 Key Laboratory for Natural Resource of Changbai Mountain & Functional Molecules, Ministry of Education, College of Pharmacy, Yanbian University, Yanji, 133002, Jilin Province, China
A sensitive analytical method was developed for the simultaneous detection of 11 chiral pharmaceuticals and their hepatic metabolites by liquid chromatography–tandem mass spectrometry (LC-MS/MS) using an ovomucoid chiral column. After optimization of the LC conditions, all pharmaceuticals examined were enantio-separated with Rs of >0.82 in LC-MS/MS analysis. The limit of detections of all pharmaceuticals by MS/MS detection ranged from 1.2 to 92.3 nM, which is approximately 1000 – 25000 times lower than those obtained by UV detection. From hepatic metabolite analyses in P450-expressing cells, metabolites of three pharmaceuticals were detected and enantio-separated. By using the proposed method, changes in the optical isomer ratio of the hepatic metabolites chlorpheniramine and verapamil caused by differential cytochrome P450 enzyme expression for each isomer, could be successfully traced.