NITCD: A streamlined metabolomics strategy based on bromine isotopes and MS-TDF software.

Area of Science:

• Metabolomics
• Analytical Chemistry
• Biochemistry

Background:

• Derivatization methods, especially stable isotope derivatization, are crucial for improving analyte coverage and accuracy in metabolomics.
• Previous homolog derivatization with MS-TDF software offered low-cost metabolomics but suffered from high false positive rates due to differential derivatization efficiencies.
• Limitations in existing methods necessitate novel approaches for enhanced metabolite identification and quantification.

Purpose of the Study:

• To introduce a novel natural isotope triple-dimensional combinatorial derivatization (NITCD) strategy to overcome limitations of previous methods.
• To enhance metabolite identification accuracy and enable reliable relative quantification in metabolomics.
• To apply the NITCD strategy in a study of rhein treatment for inflammatory bowel disease (IBD).

Main Methods:

• Development and application of the natural isotope triple-dimensional combinatorial derivatization (NITCD) strategy.
• Utilizing 4-bromo-2-hydrazinopyridine for a characteristic 1:1 isotopic doublet pattern (79Br/81Br).
• Employing mass spectrometry triple-dimensional derivatization filter (MS-TDF) software for intelligent metabolite identification.
• Using 2-hydrazinopyridine as a structurally matched internal standard for improved quantification.

Main Results:

• The NITCD strategy successfully reduced false-positive identifications and enabled reliable relative quantification.
• Application to rhein treatment in IBD detected 564 compounds in mouse plasma, 148 in colon, and 81 in spleen.
• Identified dynamic metabolic changes during rhein treatment, revealing potential reversal of IBD-induced alterations in key metabolic pathways.

Conclusions:

• The NITCD strategy offers a significant advancement in metabolomics, improving accuracy and reliability.
• This method is effective for identifying and quantifying metabolites in complex biological samples, such as plasma and tissues.
• Rhein treatment shows potential to modulate IBD-related metabolic disruptions in pathways like arachidonic acid and tryptophan metabolism.