DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells

Li Liu; Yu-Han Lu; Min-Dan Wang; Qun-Fei Zhao; Xiu-Ping Chen; Hang Yin; Chen-Guo Feng; Fang Zhang

doi:10.1016/j.jpha.2023.08.016

Volume 13 Issue 11

Nov. 2023

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Li Liu, Yu-Han Lu, Min-Dan Wang, Qun-Fei Zhao, Xiu-Ping Chen, Hang Yin, Chen-Guo Feng, Fang Zhang. DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells[J]. Journal of Pharmaceutical Analysis, 2023, 13(11): 1365-1373. doi: 10.1016/j.jpha.2023.08.016

Citation:

Li Liu, Yu-Han Lu, Min-Dan Wang, Qun-Fei Zhao, Xiu-Ping Chen, Hang Yin, Chen-Guo Feng, Fang Zhang. DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells[J]. Journal of Pharmaceutical Analysis, 2023, 13(11): 1365-1373. doi: 10.1016/j.jpha.2023.08.016

Citation:

Li Liu, Yu-Han Lu, Min-Dan Wang, Qun-Fei Zhao, Xiu-Ping Chen, Hang Yin, Chen-Guo Feng, Fang Zhang. DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells[J]. Journal of Pharmaceutical Analysis, 2023, 13(11): 1365-1373. doi: 10.1016/j.jpha.2023.08.016

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DMMIC derivatization-assisted liquid chromatography-mass spectrometry method for metabolite profiling of the glutathione anabolic pathway in esophageal cancer tissues and cells

doi: 10.1016/j.jpha.2023.08.016

Li Liu^a,b,
Yu-Han Lu^a,c,
Min-Dan Wang^a,
Qun-Fei Zhao^a,
Xiu-Ping Chen^{a
,
,},
Hang Yin^{b
,
,},
Chen-Guo Feng^{a
,
,},
Fang Zhang^{a
,
,}

a. 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;
b. Department of Thoracic Surgery, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 200127, China;
c. School of Public Health, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China

Funds:

We thank the Shanghai Municipal Committee of Science and Technology (Grant Nos.: 20XD1423400, 23ZR1460900 and 20DZ2201100), Shanghai Municipal Health Commission/Shanghai Municipal Administration of Traditional Chinese Medicine (Grant No.: ZY(2021–2023)-0501), Shanghai Science and Technology Development Fund from Central Leading Local Government (Grant No.: YDZX20223100001004), National Natural Science Foundation of China (Grant No.: 21672249), and Expenditure Budget Program of Shanghai University of Traditional Chinese Medicine (Grant Nos.: 2020LK051, and 2021LK001).

Received Date: Apr. 24, 2023
Accepted Date: Aug. 22, 2023
Rev Recd Date: Aug. 12, 2023
Publish Date: Sep. 22, 2023

Abstract

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.
- Glutathione anabolic pathway,
- Metabolite profiling,
- DMMIC derivatization,
- LC-MS,
- Esophageal squamous cell carcinoma,
- p-Hydroxycinnamaldehyde,
- KYSE-150cell

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References(40)

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