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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
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 (DMM...
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Published in: | Journal of pharmaceutical analysis 2023-11, Vol.13 (11), p.1365-1373 |
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Main Authors: | , , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | 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.
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•An effective DMMIC derivatization-assisted LC‒MS method was developed for metabolite profiling of the glutathione anabolic pathway (GAP) in complex biological samples, making it feasible to simultaneously and accurately quantify the 13 main metabolites on the GAP.•Sulfhydryl metabolites and oligopeptides with poor stability were stabilized by an addition of a reductant DTT and directly homogenizing tissue samples after thawing.•Six metabolites on the GAP including glutamine, cysteine, glycine, homocysteine, serine, and methionine were significant differences between the carcinoma and para-carcinoma tissues of esophageal squamous cell carcinoma.•p-Hydroxycinnamaldehyde, as an important active component of Momordica cochinchinensis, promoted the levels of GC and GSH on the GAP in the KYSE-150 esophageal cancer cells through affecting the expression of GCLC and GCLM genes. |
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ISSN: | 2095-1779 2214-0883 |
DOI: | 10.1016/j.jpha.2023.08.016 |