Development of a metabolomics-based data analysis approach for identifying drug metabolites based on high-resolution mass spectrometry

Hsiao-Hsien Ting ^a,1, Yi-Shiou Chiou ^b,1, Tien-Yi Chang ^c, Guan-Yu Lin ^d, Pei-Jhen Li ^d,*, Chia-Lung Shih ^c,**

^a Department of Anesthesiology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi City 60002, Taiwan

^b Master Degree Program in Toxicology, College of Pharmacy, Kaohsiung Medical University, Kaohsiung 80756, Taiwan

^c Clinical Research Center, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi City 60002, Taiwan

^d Department of Chemistry and Biochemistry, National Chung Cheng University, Chiayi 621301, Taiwan

A metabolomics-based approach to data analysis is required for drug metabolites to be identified quickly. This study developed such an approach based on high-resolution mass spectrometry. Our approach is a two-stage one that combines a time-course experiment with stable isotope tracing. Pioglitazone (PIO) was used to improve glycemic management for type 2 diabetes mellitus. Consequently, PIO was taken as a model drug for identifying metabolites. During Stage I of data analysis, 704 out of 26626 ions exhibited a positive relationship between ion abundance ratio and incubation time in a time-course experiment. During Stage II, 25 isotope pairs were identified among the 704 ions. Among these 25 ions, 18 exhibited a dose-response relationship. Finally, 14 of the 18 ions were verified to be PIO structure-related metabolite ions. Otherwise, orthogonal partial least squares-discriminant analysis (OPLS-DA) was adopted to mine PIO metabolite ions, and 10 PIO structure-related metabolite ions were identified. However, only four ions were identified by both our developed approach and OPLS-DA, indicating that differences in the designs of metabolomicsbased approaches to data analysis can result in differences in which metabolites are identified. A total of 20 PIO structure-related metabolites were identified by our developed approach and OPLS-DA, and six metabolites were novel. The results demonstrated that our developed two-stage data analysis approach can be used to effectively mine data on PIO metabolite ions from a relatively complex matrix.

Keywords: A time-course experiment, OPLS-DA, Pioglitazone, Stable isotope tracing

https://doi.org/10.38212/2224-6614.3451