DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells
10.1016/j.jpha.2023.08.016
- Author:
Li LIU
1
;
Yu-Han LU
;
Min-Dan WANG
;
Qun-Fei ZHAO
;
Xiu-Ping CHEN
;
Hang YIN
;
Chen-Guo FENG
;
Fang ZHANG
Author Information
1. The Research Center of Chiral Drugs,Shanghai Frontiers Science Center for TCM Chemical Biology,Innovation Research Institute of Traditional Chinese Medicine,Shanghai University of Traditional Chinese Medicine,Shanghai,201203,China;Department of Thoracic Surgery,Renji Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai,200127,China
- Keywords:
Glutathione anabolic pathway;
Metabolite profiling;
DMMIC derivatization;
LC-MS;
Esophageal squamous cell carcinoma;
p-Hydroxycinnamaldehyde;
KYSE-150cell
- From:
Journal of Pharmaceutical Analysis
2023;13(11):1365-1373
- CountryChina
- Language:Chinese
-
Abstract:
In this work,a new pyrylium derivatization-assisted liquid chromatography-mass spectrometry(LC-MS)method was developed for metabolite profiling of the glutathione anabolic pathway(GAP)in cancer tissues and cells.The pyrylium salt of 6,7-dimethoxy-3-methyl isochromenylium tetrafluoroborate(DMMIC)was used to label the amino group of metabolites,and a reductant of dithiothreitol(DTT)was employed to stabilize the thiol group.By combining DMMIC derivatization with LC-MS,it was feasible to quantify the 13 main metabolites on the GAP in complex biological samples,which had good linearity(R2=0.9981-0.9999),precision(interday precision of 1.6％-19.0％and intraday precision of 1.4％-19.8％)and accuracy(83.4％-115.7％).Moreover,the recovery assessments in tissues(82.5％-107.3％)and in cells(98.1％-118.9％)with GSH-13C2,15N,and Cys-15N demonstrated the reliability of the method in detecting tissues and cells.Following a methodological evaluation,the method was applied successfully to investigate difference in the GAP between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma(ESCC)and the effect of p-hydroxycinnamaldehyde(CMSP)on the GAP in KYSE-150 esophageal cancer cells.The results demonstrate that the developed method provides a promising new tool to elucidate the roles of GAP in physiological and pathological processes,which can contribute to research on drugs and diseases.
