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Measurement of the 15 N/ 14 N ratio of phenylalanine in fermentation matrix by isotope ratio mass spectrometry.
Biotechnology Letters 2017 December
OBJECTIVES: To determine the origin of 15 N-labeled phenylalanine in microbial metabolic flux analysis using 15 N as a tracer, a method for measuring phenylalanine δ15 N using HPLC coupled with elemental analysis-isotope ratio mass spectrometry (EA-IRMS) was developed.
RESULTS: The original source of the 15 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 15 N-labeled inorganic nitrogen source, rather than 15 N-labeled amino acids, was explored using this method.
CONCLUSIONS: The method described here can also be applied to the analysis of metabolic flux.
RESULTS: The original source of the 15 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 15 N-labeled inorganic nitrogen source, rather than 15 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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