气相色谱-燃烧-同位素比值质谱法分析氨基酸氮稳定同位素并初步评估水生生物体营养级

引用本文: 赵晶晶, 张忠义, 郑能建, 田晶, 朱光旭, 肖化云. 气相色谱-燃烧-同位素比值质谱法分析氨基酸氮稳定同位素并初步评估水生生物体营养级[J]. 分析化学, 2017, 45(3): 309-315. doi: 10.11895/j.issn.0253-3820.160594 shu

Citation: ZHAO Jing-Jing, ZHANG Zhong-Yi, ZHENG Neng-Jian, TIAN Jing, ZHU Guang-Xu, XIAO Hua-Yun. Stable Nitrogen Isotope Analysis of Amino Acids by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry for High-Resolution Trophic Level Estimation[J]. Chinese Journal of Analytical Chemistry, 2017, 45(3): 309-315. doi: 10.11895/j.issn.0253-3820.160594 shu

气相色谱-燃烧-同位素比值质谱法分析氨基酸氮稳定同位素并初步评估水生生物体营养级

1.
中国科学院地球化学研究所环境地球化学国家重点实验室, 贵阳 550081;
2.
中国科学院大学, 北京 100049
基金项目:
本文系国家自然科学基金项目(Nos.41425014，2016YFA061000)资助

摘要: 氨基酸稳定氮同位素(δ¹⁵N)分析能准确有效地评估生物体的营养级以及氮在食物链中的流动。本研究优化了氨基酸氮同位素的分析方法：样品在酸性条件下水解后，释放出的蛋白质氨基酸经阳离子交换树脂纯化后，衍生为对应的N-新戊酰基，O-异丙醇(N-pivaloyl-isopropyl，NPP)酯，利用气相色谱-燃烧-同位素比值质谱仪(Gas chromatography-combustion-isotope ratio mass spectrometry，GC-C-IRMS)测定其δ¹⁵N。经非极性气相色谱柱DB-5ms分离后，13种氨基酸NPP酯衍生物均可得到良好的基线分离。在样品量不低于20 ng N条件下，GC-C-IRMS方法的精密度优于1‰，测得的δ¹⁵N值与EA-IRMS法测得的δ¹⁵N值没有明显差异。阳离子树脂纯化前后各氨基酸δ¹⁵N值差异低于1‰，表明没有产生明显的同位素分馏。采用本方法成功地估算了阿哈湖生态系统中常见水生生物的营养级，可作为研究氨基酸代谢以及生态系统特征的新方法。

关键词:

English

Stable Nitrogen Isotope Analysis of Amino Acids by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry for High-Resolution Trophic Level Estimation

1.
State Key Laboratory of Environmental Geochemistry, Institute of Geochemistry, Chinese Academy of Sciences, Guiyang 550001, China;
2.
University of Chinese Academy of Sciences, Beijing 100049, China
Received Date: 09 August 2016
Revised Date: 20 December 2016
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 41425014, 2016YFA0601000)

Abstract: The analysis of stable nitrogen isotopic composition (δ¹⁵N) of individual amino acid was recognized as an effective method for estimating the trophic level of organisms and detecting the nitrogen flow in food webs. In this study, we evaluated a two-stage procedure of esterification followed by acylation, in which biological samples underwent hydrolysis in acid and the released individual amino acids were derivative into the corresponding N-pivaloyl-iso-propyl (NPP) esters for gas chromatography-combustion-isotope ratio mass spectrometric (GC-C-IRMS) analysis. A total of 13 kinds of individual amino acid derivatives were baseline separated on a nonpolar gas chromatography column (DB-5ms). The amount of sample for each test was not less than 20 ng N on column. High correlations were observed between the δ¹⁵N values respectively obtained by GC-C-IRMS and element analysis-isotope ration mass spectrometry (EA-IRMS). Furthermore, the mean precision of this method was better than 1‰. Cation-exchange chromatograph was used to purify the samples, and the difference of the detection δ¹⁵N values before and after purification by the resin was within 1‰. This method was applied to estimate the trophic level of various natural freshwater organisms from Aha Lake. The present study provided a new idea for the application of stable nitrogen isotope (δ¹⁵N) in the trophic level estimation of organisms and metabolism analysis of amino acid.

Key words:

N-pivaloyl-iso-propyl esters
/ Amino acid
/ Gas chromatography-combustion-isotope ratio mass spectrometry
/ Trophic level

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

气相色谱-燃烧-同位素比值质谱法分析氨基酸氮稳定同位素并初步评估水生生物体营养级

English

Stable Nitrogen Isotope Analysis of Amino Acids by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry for High-Resolution Trophic Level Estimation

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

气相色谱-燃烧-同位素比值质谱法分析氨基酸氮稳定同位素并初步评估水生生物体营养级

English

Stable Nitrogen Isotope Analysis of Amino Acids by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry for High-Resolution Trophic Level Estimation

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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通讯作者: 陈斌, bchen63@163.com

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