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利用气相色谱-质谱联机测定土壤氨基酸手性异构体13C富集比例

何红波1;张威1,2;丁雪丽1,2;白震1,2;刘宁1,2张旭东1   

  1. 1中国科学院沈阳应用生态研究所陆地生态过程重点实验室, 沈阳 110016;2中国科学院研究生院, 北京 100039
  • 收稿日期:2007-06-26 修回日期:1900-01-01 出版日期:2008-06-20 发布日期:2008-06-20

Determination of 13C-enrichment in soil amino acid enantiomers by gas chromatogram/mass spectrometry.

HE Hong-bo1;ZHANG Wei1,2;DING Xue-li1,2;BAI Zhen1,2;LIU Ning1,2;ZHANG Xu-dong1   

  1. 1Key Laboratory of Terrestrial Ecological Process, Institute of Applie
    d Ecology, Chinese Academy of Sciences, Shenyang 110016, China;2Graduate
    University of Chinese Academy of Sciences, Beijing 100039, China
  • Received:2007-06-26 Revised:1900-01-01 Online:2008-06-20 Published:2008-06-20

摘要: 氨基酸手性异构体的转化与更新程度在表征土壤有机质的循环转化机制方面具有重要意义.为有效区分土壤中原有的和利用外加碳源新合成的氨基酸,本文建立了稳定同位素培养气相色谱/质谱联机测定土壤氨基酸手性异构体13C富集比例的方法.由于培养过程中加入的13C葡萄糖被迅速利用形成氨基酸碳骨架,因而利用质谱技术可检测同位素的富集强度的变化.外加葡萄糖直接合成氨基酸的比例可利用质谱碎片(F+n)的相对强度变化来评价(n为质谱碎片F中含有的碳原子数目);而源于葡萄糖的13C同位素在氨基酸分子中的富集程度是所有同位素峰相对强度变化的总和.13C在目标化合物中的富集比例用原子百分超(APE)评价,D氨基酸的APE还需进一步利用水解诱导的外消旋化系数校正.13C在目标化合物中的富集程度可反映各氨基酸手性异构体的碳循环速率大小,是研究土壤氨基酸动态变化的有力工具.

关键词: 油松人工林, 近自然经营, 幼苗更新, 林分生长, 物种多样性, 土壤特性

Abstract: The transformation and renewal of amino acid enantiomers is of significance in indicating the turnover mechanism of soil organic matter. In this paper, a method of gas chromatogram/mass spectrometry combined with U-13 -C-glucose incubation was developed to determine the 13C-enrichment in soil amino acid enantiomers, which could effectively differentiate the original and the newly synthesized amino acids in soil matrix. The added U-13-C-glucose was utilized rapidly to structure the amino acid carbon skeleton, and the change of relative abundance of isotope ions could be determined by mass spectrometry. The direct incorporation of U-13C glucose was estimated by the intensity increase of m/z (F+n) to F (F was parent fragment, and n was the carbon number in the fragment), while the total isotope incorporation from the added 13C could be calculated according to the abundance ratio increment summation from m/z (Fa+1) through (Fa+T) (Fa was the fragment containing all original skeleton carbons, and T was the carbon number in the amino acid molecule). The 13C enrichment in the target compound was expressed as atom percentage excess (APE), and that of D-amino acid needed to be corrected by the coefficient of hydrolysisinduced racemization. The13C enrichment reflected the carbon turnover velocity of individual amino acid enantiomers, and was powerful to investigate the dynamics of soil amino acids.

Key words: Pinus tabulaeformis plantation, closetonatural forest management, seedling regeneration, stand growth, species diversity, soil characteristics.