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[前沿] 基于氯甲酸酯的氨基酸化合物碳、氮同位素分析

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发表于 2014-3-28 01:30 | 显示全部楼层 |阅读模式
Compound-specific δ13C and δ15N analysis of amino acids: a rapid, chloroformate-based method for ecological studies
Abstract
RATIONALE
Compound-specific stable isotope analysis of amino acids has proven informative to many ecological systems, but only a handful of analytical methods are routinely employed. We evaluated a simple, rapid procedure in which biological samples undergo short-duration acid hydrolysis and the resulting amino acids are derivatized with methyl chloroformate for gas chromatography/combustion/isotope-ratio mass spectrometry (GC/C/IRMS).

METHODS
Amino acid derivatives were separated on a polar gas chromatography column, combusted, and δ13C and δ15N values were measured. Tests of reproducibility and accuracy were conducted for amino acid reference mixtures and biological samples. A brief case study of turtles was used to assess whether isotopic data were consistent with a priori ecological expectations.

RESULTS
The methyl chloroformate based reaction successfully converted 15 amino acids from acid hydrolysates of biological materials into separable derivatives. The δ13C and δ15N values had high average measurement precision (σ <1 ‰). Reference materials were measured accurately, with good agreement between EA/IRMS and GC/C/IRMS determinations. Analysis of turtle blood samples yielded data consistent with their trophic ecology.

CONCLUSIONS
This derivatization method is a rapid means of determining carbon and nitrogen isotopic ratios of amino acids present in the biological materials often sampled for ecological studies. While amino acids with charged or polar side chains do not have uniformly high recoveries, the average precision of measurements is comparable with that of other, more established methods. Batches of samples may be prepared from many raw materials in less than a day, representing a significant reduction in preparation time over prevailing methods. Copyright © 2013 John Wiley & Sons, Ltd.

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procedure, 氨基酸, 碳同位素, 氮同位素, 气相色谱
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