气相色谱-质谱联用法检测13C标记葡萄糖的同位素丰度

doi:10.7538/tws.2020.33.05.0286

摘要/Abstract

摘要：

为了评价稳定同位素¹³C标记葡萄糖作为示踪工具的质量，本研究通过衍生化试剂的筛选、衍生化条件的优化以及选择合适的质谱采集模式，建立基于气相色谱质谱联用（GC-MS）技术测定¹³C标记葡萄糖同位素丰度的方法。除衍生条件的优化外，本文重点通过配制不同浓度的葡萄糖溶液对两种不同采集模式的检测结果准确性进行探究。结果表明，衍生化试剂N-甲基二(三氟乙酰胺)（MBTFA）的衍生效果较N-甲基-N-(三甲基硅烷基)三氟乙酰胺（MSTFA）更优，在低浓度下采用离子扫描模式（SIM）检测结果的准确性高。本研究的¹³C标记葡萄糖同位素丰度检测方法，其精密度优于0.2%，绝对偏差小于0.3%，可作为¹³C标记葡萄糖同位素丰度的检测方法，并为相关领域研究提供技术支持。

关键词: 气相色谱-质谱法, 衍生化, 葡萄糖, 同位素丰度

Abstract:

Glucose labeled with stable isotope ¹³C is a commonly used tracing tool, which have been applied in life sciences and other fields. In the evaluation of tracer tools, isotope abundance is a key indicator of tracer. In this paper, a method for the determination of ¹³C labeled glucose abundance based on gas chromatographymass spectrometry (GC-MS) technology is established. The isotope abundance of ¹³C labeled glucose was detected through the screening of derivatization reagents, the optimization of derivatization conditions and the selection of appropriate mass spectrometry acquisition mode. Through the preparation of different concentrations of glucose solution to explore the accuracy of the two acquisition modes, it was found that the accuracy of detection results can be greatly improved by using ion scanning mode (SIM) at low concentration. In addition, it was also found that the derivatization reagent N-methyl bis (trifluoroacetamide) (MBTFA) has a better derivatization effect than N-Methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA). The calculated value of the established ¹³C labeled glucose detection method was basically consistent with the theoretical value, the relative standard deviations (RSD, n=8) were below 0.2%，and absolute deviation were below 0.3%. The method for isotope abundance detection is accurate and can provide ¹³C labeled glucose isotope abundance detection method and provide technical support for research in related fields.

Key words: GC-MS, derivatization, glucose, isotope abundance

陈燕秋;侯捷;雷雯;李良君;徐仲杰;高慧敏. 气相色谱-质谱联用法检测¹³C标记葡萄糖的同位素丰度[J]. 同位素, 2020, 33(5): 286-292.

CHEN Yanqiu;HOU Jie;LEI Wen;LI Liangjun;XU Zhong jie;GAO Huimin. Determination of Isotope Abundance of ¹³C-labeled Glucose by Gas Chromatography-Mass Spectrometry[J]. Journal of Isotopes, 2020, 33(5): 286-292.

参考文献

［1］Caron A, Lavoie C, Francois C, et al. Oxidation of ［C］ glucose ingested before and/or during prolonged exercise［J］. European Journal of Applied Physiology, 2003(2): 217-223.
［2］王龙. 基于13C同位素标记实验龟裂链霉菌代谢通量分析方法的建立与应用［D］. 上海：华东理工大学,2013.
［3］Peronnet F, Abdelaoui M, Lavoie C, et al. Effect of a 20-day ski trek on fuel selection during prolonged exercise at low workload with ingestion of glucose-¹³C［J］. European Journal of Applied Physiology, 2009(1): 41-49.
［4］Shew S B, Keshen T H, Jahoo R F, et al. Assessment of cysteine synthesis in very low birth weight neonates using a［13C6］ glucose tracer［J］. J Pediatr Sugr, 2005, 40(1): 52-56.
［5］Lefebvre P, Luyckx A, Mosora F, et al. Effect of butylbiguanide therapy in obese mildly diabetic subjects［J］. Diabetologia, 1978(1): 39-45.
［6］张敏,管樑,朱承谟,等. 国产¹³C-葡萄糖呼气试验检测糖尿病小鼠胰岛素抵抗的实验研究［J］. 上海交通大学学报(医学版),2012,32(4):463-466.Zhang Min, Guan Liang, Zhu Chengmo, et al. Experimental study on domestic ¹³C-glucose breath test to detect insulin resistance in diabetic mice［J］. Journal of Shanghai Jiaotong University (Medical Edition), 2012, 32(4): 463-466（in Chinese）.
［7］Vonk R J, Stellard F, Priebe M G, et al. The ¹³C/²H-glucose test for determination of small intestinal lactase activity［J］. European Journal of Clinical Investigation, 2001, 1(3): 226-233.
［8］王建,王泽建,黄明志,等. ¹³CMFA 过程中GC-MS分析菌体蛋白氨基酸的¹³C标记丰度［J］. 中国生物工程杂志,2009,29(7):87-93.Wang Jian, Wang Zejian, Huang Mingzhi, et al. Analysis of ¹³C labeled abundance of amino acids in bacterial proteins by GC-MS during ¹³C MFA process［J］. Chinese Journal of Bioengineering, 2009, 29(7): 87-93（in Chinese）.
［9］Tissot S, Normand S, Guilluy R, et al. Use of a new gas chromatograph isotope ratio mass spectrometer to trace exogenous ¹³C labelled glucose at a very low level of enrichment in man［J］. Diabetologia, 1990(8): 449-456.
［10］徐仲杰,卢伟京,卢浩. ¹³C₆-1,2,5,6-O-双异丙叉-α-D-呋喃葡萄糖的合成［J］. 同位素,2011,24(4):229-233.Xu Zhongjie, Lu Weijing, Lu Hao. Synthesis of ¹³C₆-1,2,5,6-O-bisisopropylidene-α-D-furan glucose［J］. Journal of Isotopes, 2011, 24(4): 229-233（in Chinese）.
［11］Qi Chao, Liu Liyan. Enzymatic synthesis of large amounts of ¹³C-labeled uridine diphosphoglucose［J］. Journal of University of Science and Technology of China, 2004, 34(5): 624-629.
［12］张亮,杜晓宁,李良君. L-亮氨酸-U-¹³C₆的研制［J］. 同位素,2011,24(增刊):106-111.Zhang Liang, Du Xiaoning, Li Liangjun. Preparation of L-Leucine-U-¹³C₆［J］. Journal of Isotopes, 2011, 24 (Suppl): 106-111（in Chinese）.
［13］王静,陈悦,袁子民,等. 基于尿液代谢组学分析肉豆蔻麸煨炮制前后对大鼠长期毒性的作用差异［J］. 中国实验方剂学杂志,2018,24(4):8-13.Wang Jing, Chen Yue, Yuan Zimin, et al. Analysis of the difference in the long-term toxicity of rat nutmeg bran before and after processing based on urine metabolomics［J］. Chinese Journal of Experimental Pharmacology, 2018, 24(4): 8-13（in Chinese）.
［14］Rhee Eugene P, Gerszten Robert E. Metabolomics and cardiovascular biomarker discovery［J］. Clinical chemistry, 2012, 58(1): 139-147.
［15］Boerboom Gavin, van Kempen Theo, Navarro-Villa Alberto, Pérez-Bonilla Adriano. Unraveling the cause of white striping in broilers using metabolomics［J］. Poultry science, 2018, 97(11): 1-10.
［16］Chen M, Lu B, Li Y, et al. Metabolomics insights into the modulatory effects of long-term compound polysaccharide intake in high-fat diet-induced obese rats［J］. Nutrition & Metabolism, 2018, 15(1): 8.
［17］Gay L J, Schneiter P, Schutz Y, et al. A non- invasive assessment of hepatic glycogen kinetics and postabsorptive gluconeogenesis in man［J］. Diabetologia, 1994, 37(5): 517-523.
［18］Yatscoff R W, Foster R T, Aspeslet L J, et al. ¹³C glucose breath test for the diagnosis of diabetes: WO, 1999056790［P］. 1999-02-12.
［19］Riddell M C, Baror O, Schwarcz H P, et al. Substrateutilization in boys during exercise with ［¹³C］-glucose ingestion［J］. European Journal of Applied Physiology, 2000(4): 441-448.
［20］杨子昂,吴国豪,吴肇汉. 稳定核素示踪法应用于内毒素诱导兔高代谢脓毒血症的代谢研究［C］∥全国肠外肠内营养学术会议. 合肥：中华医学会，2010.
［21］侯捷,张鹏帅,侯静华,等. 茶叶中多种氨基酸¹⁵N同位素丰度的检测研究［J］. 同位素,2018,31(4):199-207.Hou Jie, Zhang Pengshuai, Hou Jinghua, et al. Detection of ¹⁵N isotope abundance of various amino acids in tea［J］. Journal of Isotopes, 2018, 31(4): 199-207（in Chinese）.
［22］雷雯,侯捷,杜晓宁,等. 血液中¹⁵N标记氨基酸同位素丰度［J］. 分析化学,2015,43(10):1465-1471.Lei Wen, Hou Jie, Du Xiaoning, et al. Determination of isotope abundance of ¹⁵N labeled amino acids by gas chromatography-Mass spectrometry［J］. Analytical Chemistry, 2015, 43(10): 1465-1471（in Chinese）.
［23］侯捷,雷雯,杜晓宁,等. 气相色谱-质谱联用法分析15N标记精氨酸发酵液中的氨基酸组分及其同位素丰度［J］. 分析测试学报,2016,35(9):1132-1136. Hou Jie, Lei Wen, Du Xiaoning, et al. Analysis of amino acid components and isotopic abundance in ¹⁵N-labeled arginine fermentation broth by gas chromatography-mass spectrometry［J］. Journal of Analysis and Testing, 2016, 35(9): 1132-1136（in Chinese）.

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气相色谱-质谱联用法检测¹³C标记葡萄糖的同位素丰度

Determination of Isotope Abundance of ¹³C-labeled Glucose by Gas Chromatography-Mass Spectrometry

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