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Selective and Sensitive Determination of Protein and Non‐Protein Amino Acids by Capillary Gas Chromatography with Nitrogen–Phosphorus Selective Detection
Author(s) -
Kataoka Hiroyuki
Publication year - 1997
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/(sici)1099-0801(199705)11:3<154::aid-bmc652>3.0.co;2-g
Subject(s) - chemistry , chromatography , amino acid , gas chromatography , urine , calibration curve , nitrogen , phosphorus , detection limit , organic chemistry , biochemistry
A selective and sensitive method for the determination of protein and non‐protein amino acids by capillary gas chromatography (GC) has been developed. The amino acids were converted into their N ( O,S )‐isobutoxycarbonyl methyl ester derivatives and measured by GC with nitrogen‐phosphorus selective detection using a DB‐17ht capillary column. Using this method, the derivatives of the 21 protein amino acids and the 33 non‐protein amino acids provided excellent NPD responses, and were quantitatively and reproducibly resolved within 28 min. The detection limits of these amino acids were 6–150 pg per injection. The calibration curves were linear in the range 0.02–2 μg for each amino acid, the correlation coefficients being above 0.990. This method was successfully applied to small urine samples without prior clean‐up, and analysed without any influence from coexisting substances. Overall recoveries of amino acids added to urine sample were 83–108%. The analytical results of free amino acid contents in urine samples of normal subjects are presented. © 1997 John Wiley & Sons, Ltd.