Polyamine quantitation by LC-MS using isobutyl chloroformate derivatives.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Metabolomics

Background:

• Polyamines are crucial metabolites often inadequately quantified in standard metabolomics workflows.
• Existing methods lack comprehensive coverage or require specialized techniques for polyamine analysis.

Purpose of the Study:

• To develop and validate a straightforward protocol for the quantitative analysis of polyamines.
• To enable polyamine measurement as a standalone or supplementary step in existing metabolomics pipelines.

Main Methods:

• Isobutyl-chloroformate derivatization of metabolite extracts.
• Quantitative analysis using a 15-minute Liquid Chromatography-Mass Spectrometry (LC-MS) method.
• Utilized triple quadrupole mass spectrometry with at least two transitions per compound.

Main Results:

• Successfully quantified thirteen polyamines and two internal standards.
• Achieved individual quantification of co-eluting isomers N1- and N8-acetylspermidine.
• Defined the linear dynamic range for each quantified compound across various biological sample types.

Conclusions:

• The developed isobutyl-chloroformate derivatization method offers a simple, robust, and rapid approach for polyamine quantification.
• This protocol significantly improves polyamine coverage in metabolomics, applicable both independently and post-initial analysis.
• The method facilitates detailed polyamine profiling, including isomer-specific analysis, in diverse biological matrices.