Targeted profiling of polar metabolites in cancer metabolic reprogramming by hydrophilic interaction liquid chromatography-tandem mass spectrometry
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Updated: Sep 22, 2026
A Strategy for Sensitive, Large Scale Quantitative Metabolomics
Published on: May 27, 2014
Xue-Man Dong1, Pu Wu2, Long-Hui Cheng2
1Innovation Research Institute of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; School of Pharmacy, Hangzhou Normal University, Hangzhou 311121, China; Key Laboratory of Elemene Class Anti-Cancer Chinese Medicines, Engineering Laboratory of Development and Application of Traditional Chinese Medicines, Collaborative Innovation Center of Traditional Chinese Medicines of Zhejiang Province, Hangzhou Normal University, Hangzhou 311121, China.
Abstract:
Metabolic reprogramming of cancer cells is a hallmark of cancer, in which the polar metabolites involving aerobic glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, and glutaminolysis play a crucial role in the occurrence and development of cancer. Therefore, targeted analysis of the polar metabolites in these pathways is of great value for understanding cancers, finding diagnostic biomarkers, and identifying therapeutic targets. However, it is still challenging to directly determine polar metabolites in these pathways without derivatization due to their diverse chemical properties, isomers, and strong polarity. Herein, a highly selective and sensitive HILIC-MS/MS method was developed for direct determination of the polar metabolites in aerobic glycolysis, PPP, TCA cycle, and glutaminolysis pathways. Without derivatization, 19 polar metabolites and their isomers with carbonyl, carboxyl, or phosphoryl groups in human plasma and cell extracts of prostate cancer (PC) were determined with strong retention and high resolution. This method has been widely verified by measuring linearity, precision, sensitivity, repeatability, matrix effect, and accuracy. The analysis of plasma samples by HILIC-MS/MS revealed distinct PC-specific metabolic signatures compared to a healthy control. In addition, this method could also be used to screen the targets of metabolic inhibitors at the cellular level. We conclude that the developed HILIC-MS/MS method provides a valuable means to study the cancer metabolic reprogramming or energy metabolism in living organisms.
Insights
A new HILIC-MS/MS method enables direct analysis of polar metabolites in key cancer pathways. This technique aids in understanding cancer metabolism, identifying biomarkers, and screening drug targets in prostate cancer.
Area of Science:
- Metabolomics
- Cancer Biology
- Analytical Chemistry
Background:
- Metabolic reprogramming is a hallmark of cancer, involving pathways like aerobic glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, and glutaminolysis.
- Polar metabolites in these pathways are crucial for cancer occurrence and development, making their targeted analysis valuable for understanding cancer, biomarker discovery, and therapeutic target identification.
- Direct determination of polar metabolites is challenging due to diverse chemical properties, isomers, and strong polarity, often requiring derivatization.
Purpose of the Study:
- To develop a highly selective and sensitive HILIC-MS/MS method for the direct determination of polar metabolites in key cancer metabolic pathways without derivatization.
- To apply this method for analyzing polar metabolites in human plasma and prostate cancer cell extracts.
- To investigate the utility of the method for identifying cancer-specific metabolic signatures and screening metabolic inhibitor targets.
Main Methods:
- Development and validation of a Hydrophilic Interaction Liquid Chromatography-tandem Mass Spectrometry (HILIC-MS/MS) method.
- Direct determination of 19 polar metabolites and their isomers in human plasma and prostate cancer cell extracts.
- Validation of the method for linearity, precision, sensitivity, repeatability, matrix effect, and accuracy.
Main Results:
- The HILIC-MS/MS method achieved strong retention and high resolution for direct determination of polar metabolites and their isomers without derivatization.
- Distinct prostate cancer-specific metabolic signatures were identified in plasma samples compared to healthy controls.
- The method demonstrated applicability in screening targets of metabolic inhibitors at the cellular level.
Conclusions:
- The developed HILIC-MS/MS method provides a valuable tool for the direct and sensitive analysis of polar metabolites involved in cancer metabolic reprogramming.
- This method facilitates a deeper understanding of cancer energy metabolism in biological systems.
- It offers potential for biomarker discovery and therapeutic target identification in cancer research.
