LC/MS/MS Method Package for Short Chain Fatty Acids

For the derivatization using 3-nitrophenylhydrazine, carbodiimide is used as a condensing agent, and reaction proceeds under mild conditions. Moreover, in order to efficiently carry out reactions with both short-chain fatty acids and organic acids, this method package includes derivatization using 3-NPH instead of the more commonly used 2-NPH. Also, 3-NPH derivatizations after reactions are stable, and quantitative analysis of the short-chain fatty acids or organic acids in the sample can be performed through reaction with an appropriate amount of derivatization reagent. Additionally, as reactions take place in water solutions using this derivatization reagent, the sample will not be dried and volatile components will not be lost.

Derivatization Reaction with 3-nitrophenylhydrazine (3-NPH)

The following example shows an analysis using the LCMS-8060 of a standard mixed solution of short-chain fatty acids/organic acids. An MRM chromatogram of short-chain fatty acids/organic acids derivatized using 3-NPH is shown. As can be seen, sufficient separations of the components, such as the structural isomers butyric acid, isobutyric acid, fumaric acid, maleic acid, citric acid, and isocitric acid were obtained. This method package includes MRM transitions that were formulated beforehand in order to achieve optimization for 3-NPH derivatizations for short-chain fatty acids/organic acids. As a result, short-chain fatty acids (C2 to C5) and organic acids representative of central metabolic pathways, which are difficult to analyze with LCMS using a conventional reversed phase system, can be analyzed simultaneously.

LabSolutions LCMS Ver. 5.99SP2 or later is required.

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