Objectives: To determine the origin of N-labeled phenylalanine in microbial metabolic flux analysis using N as a tracer, a method for measuring phenylalanine δN using HPLC coupled with elemental analysis-isotope ratio mass spectrometry (EA-IRMS) was developed.
Results: The original source of the N-labeled phenylalanine was determined using this new method that consists of three steps: optimization of the HPLC conditions, evaluation of the isotope fractionation effects, and evaluation of the effect of pre-processing on the phenylalanine nitrogen stable isotope. In addition, the use of a N-labeled inorganic nitrogen source, rather than N-labeled amino acids, was explored using this method.
Conclusions: The method described here can also be applied to the analysis of metabolic flux.
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