Expanding Metabolome Coverage in LC-MS/MS Analysis through Hydralazine-Based Multifunctional Derivatization

Area of Science:

• Analytical Chemistry
• Biochemistry
• Mass Spectrometry

Background:

• Metabolomics aids disease biomarker discovery and mechanism investigation.
• Challenges persist in detecting diverse metabolites due to sensitivity limitations.
• Comprehensive metabolome profiling requires improved analytical techniques.

Purpose of the Study:

• To develop a novel chemical derivatization method for sensitive detection of poorly ionized metabolites.
• To enhance the analysis of small metabolites using liquid chromatography-tandem mass spectrometry (LC-MS/MS).
• To facilitate large-scale quantitative analysis of metabolites in biological samples.

Main Methods:

• Developed a hydralazine (HZN)-based derivatization strategy targeting carbonyl, carboxyl, and phosphoryl groups.
• Optimized derivatization parameters, including condensation reagents.
• Utilized liquid chromatography-tandem mass spectrometry (LC-MS/MS) for detection and quantification.

Main Results:

• Achieved significant sensitivity improvements (10-5000-fold) for HZN-derivatized metabolites.
• Established limits of quantitation from 5 pM to 100 nM for various metabolites.
• Successfully applied the method to reveal metabolic alterations in mice on a high-fat diet.

Conclusions:

• The HZN derivatization method offers highly sensitive detection of diverse small metabolites.
• This approach simplifies MS/MS analysis and enables efficient large-scale metabolomic studies.
• The method shows promise for investigating disease-related metabolic changes, particularly those involving gut microbiota.