生物体中氨基酸单体碳稳定同位素测试方法研究  

Study on the Compound Specific Carbon Stable Isotope Determina⁃tion of Amino Acids in Organisms

作　　者：梁建鑫 尹希杰 苏静 林锡煌 李玉红 LIANG Jian-xin;YIN Xi-jie;SU Jing;LIN Xi-huang;LI Yu-hong(Information and Testing Support Center,Third Institute of Oceanography,Ministry of Natural Resources,Xiamen 361005,China;Zhoushan Field Scientific Observation and Research Station for Marine Geo-hazards,China Geological Survey,Qingdao 266237,China)

机构地区：[1]自然资源部第三海洋研究所分析测试中心,福建厦门361005 [2]中国地质调查局舟山海洋地质灾害野外科学观测研究站,山东青岛266237

出　　处：《分析测试学报》2024年第3期447-454,共8页Journal of Instrumental Analysis

基　　金：中国地质调查局舟山海洋地质灾害野外科学观测研究站开放基金资助项目(ZSORS-22-10);国家自然科学基金项目(41976050,42176057)。

摘　　要：氨基酸作为蛋白质的基本组成单位,是重要的生命物质,其单体碳同位素研究在生物地球化学、生态学、生物体代谢和环境科学等领域具有重要意义。该文优化了海参和海藻氨基酸提取和纯化流程,通过N新戊酰基-O-异丙酯(NPP)方法衍生化后,分别用气相色谱-质谱(GC-MS)和气相色谱-燃烧-同位素比值质谱(GC-C-IRMS)测试其浓度和碳同位素组成。结果显示,15种氨基酸单体的分离效果较好,回收率为46.4%~96.3%,各氨基酸在1.0~16.0µmol/L范围内线性关系良好(r^(2)为0.987~0.999)。15种氨基酸单体衍生物δ^(13)C值的标准偏差均小于0.30‰(n=10),在0.6~2.0 mmol/L浓度范围内δ^(13)C的平均误差为±0.24‰,方法检出限为0.6 nmol。海参和海藻样品各氨基酸单体δ^(13)C值的范围分别为-31.10‰~-8.58‰和-30.53‰~-13.76‰,标准偏差均在0.33‰以内,可满足生物体氨基酸单体碳同位素的测试精度需求。As the basic unit of protein,amino acids(AAs)are important living substances.The study of their monomer carbon isotope is of great significance in the fields of biogeochemistry,ecolo⁃gy,organism metabolism and environmental science.In this paper,we optimized the extraction and purification processes of AAs from sea cucumber and seaweed.After derivatization by N-pivaloyl-Oisopropyl(NPP)method,the concentration and carbon isotope composition were tested by gas chro⁃matography-mass spectrometry(GC-MS)and gas chromatography-combustion-isotope ratio mass spectrometry(GC-C-IRMS).The results showed that 15 AAs were successfully separated,the re⁃coveries were between 46.4%and 96.3%,and the linear relationship of each amino acid was good in the range of 1-16µmol/L(r^(2)=0.987-0.999).The standard deviations ofδ^(13)C values of 15 AAs mono⁃mer derivatives were less than 0.30‰(n=10),and the average error ofδ^(13)C was±0.24‰in the con⁃centration range of 0.6-2.0 mmol/L.The detection limit of this method was 0.6 nmol.Theδ^(13)C val⁃ues of AAs monomers in sea cucumber and seaweed samples ranged from-8.58‰to-31.10‰,and-13.76‰to-30.53‰,respectively,and the standard deviations were all within 0.33‰,which could meet the accuracy requirements of carbon isotope analysis of AAs monomers in organisms.

关 键 词：气相色谱-燃烧-同位素比值质谱(GC-C-IRMS) N-新戊酰基-O-异丙酯(NPP) 氨基酸 碳稳定同位素 

分 类 号：O657.63[理学—分析化学] O629.7[理学—化学]