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• 研究与进展 •    

UPLC-ESI-MS分析中棉花次生代谢物标准品推定和加合物形成

李社增1,2,牛露欣2,李博超3,陈秀叶2,马平2*,马峙英1*


1. 河北农业大学/教育部华北作物种质资源重点实验室,河北 保定 071000;2. 河北省农林科学院植物保护研究所/河北省农业有害生物综合防治工程技术研究中心/农业农村部华北北部作物有害生物综合治理重点实验室,河北 保定 071000;3. 南加州大学生物医学工程系,美国 洛杉矶 90089


第一作者简介:李社增(1966-),研究员,shezengli@163.com。*通信作者:pingma88@126.commazhiying1958@126.com

基金项目:国家重点研发计划(2017YFD0201900);国家现代农业产业技术体系—棉花产业技术体系(CARS-15-18)

  

  1. 1. 河北农业大学/教育部华北作物种质资源重点实验室,河北 保定 071000;2. 河北省农林科学院植物保护研究所/河北省农业有害生物综合防治工程技术研究中心/农业农村部华北北部作物有害生物综合治理重点实验室,河北 保定 071000;3. 南加州大学生物医学工程系,美国 洛杉矶 90089
  • 收稿日期:2020-06-12 修回日期:2020-07-23 接受日期:2020-07-23 出版日期:2020-07-27 发布日期:2020-07-27
  • 通讯作者: pingma88@126.com;mazhiying1958@126.com
  • 作者简介:李社增(1966-),研究员,shezengli@163.com
  • 基金资助:
    国家重点研发计划(2017YFD0201900);国家现代农业产业技术体系—棉花产业技术体系(CARS-15-18)

Putative Identification and Adduct Formation of Reference Materials for Cotton Secondary Metabolite using UPLC-ESI-MS Analysis

Li Shezeng1, 2, Niu Luxin2, Li Bochao3, Chen Xiuye2, Ma Ping2*, Ma Zhiying1*   

  1. 1. North China Key Laboratory for Crop Germplasm Resources of Education Ministry, Hebei Agricultural University, Baoding, Hebei 071001, China; 2. Integrated Pest Management Center of Hebei Province, Key Laboratory of IPM on Crops in Northern Region of North China, Ministry of Agriculture and Rural Affairs/Plant Protection Institute, Hebei Academy of Agricultural and Forestry Sciences, Baoding, Hebei 071000, China; 3. Department of Biomedical Engineering, University of Southern California, Los Angeles, CA 90098, USA
  • Received:2020-06-12 Revised:2020-07-23 Accepted:2020-07-23 Online:2020-07-27 Published:2020-07-27

摘要:

【目的】建立超高效液相色谱电喷雾质谱(Ultra-performance liquid chromatography-electrospray ionization-mass spectrometry,UPLC-ESI-MS)分析中棉花代谢产物的高通量鉴定方法;探讨特定UPLC和ESI模式条件下棉花次生代谢产物的加合物种类、主导加合物及适宜的ESI模式。【方法】利用高效液相色谱电喷雾质谱对18个棉花代谢产物标准品进行分析,采用在线XCMS软件进行无靶标质谱数据提取,利用MATLAB软件编程计算程序建立标准品鉴定方法。【结果】针对电喷雾正、负离子模式,建立了基于计算准确相对分子质量的棉花次生代谢产物标准品的快速鉴定方法POSid和NEGid;特定超高效液相色谱和电喷雾正、负离子模式下14个标准品得到正确鉴定。正离子模式下出现[M+H]+、[M+Na]+、[M+NH4]+、[2M+NH4]+、[2M+Na]+和[2M+H]+ 6种加合物,负离子模式下出现[M-H]-、[2M-H]-、[M+Cl]-、[M+FA-H]-、[3M-H]-、[M+Na-2H]-、[M-H2O-H]-和[M+TFA-H]- 8种加合物,单个标准品的质谱可观察到1~6种加合物,每个标准品均有主导加合物并具有电喷雾离子模式的偏好性。蜜二糖适合电喷雾正离子模式检测,棉酚适合2种离子模式检测;12种化合物均是负离子模式信号强于正离子模式,适合采用负离子模式检测。【结论】建立的基于计算准确相对分子质量的代谢产物鉴定方法,能实现对18个棉花次生代谢产物标准品的无靶标质谱数据的鉴定。特定超高效液相色谱和电喷雾条件下,棉花次生代谢产物的主导加合物具有电喷雾离子模式的偏好性。这些结果为开展棉花代谢组研究提供了技术和理论数据支撑。

关键词: 棉花次生代谢产物; 超高效液相色谱-电喷雾质谱; 质谱鉴定; 加合物

Abstract: [Objective] The aim this study is to establish an identification method for cotton metabolites in ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS) analysis, and to investigate the adduct types, dominant adducts and appropriate ESI ion modes of cotton secondary metabolites under the determined UPLC and ESI conditions. [Method] UPLC-ESI-MS was employed to analyze 18 cotton metabolite standards, Online XCMS software was used to extract the nontargeted mass spectrum data, and MATLAB software was used to prepare calculation programs for the identification method of cotton metabolite standards. [Result] A high-throughput identification method of cotton secondary metabolite, named POSid and NEGid separately for the positive and negative ESI modes, was established based on the calculated accurate molecular weight. In the determined UPLC condition and positive and negative ESI mode, 14 cotton metabolite standards were correctly identified. There found 6 adducts including [M+H]+, [M+Na]+, [M+NH4]+, [2M+NH4]+, [2M+Na]+ and [2M+H]+ in positive ion mode, and 8 adducts including [M-H]-, [2M-H]-, [M+Cl]-, [M+FA-H]-, [3M-H]-, [M+Na-2H]-, [M-H2O-H]- and [M+TFA-H]- in negative ion mode, while 1 to 6 adducts were observed in the mass spectrum of a single standard, and each metabolite standard had a dominant adduct of a preference for ESI mode. Melibose was suitable for ESI positive ion mode detection, gossypol was suitable for both ion modes detection; and 12 compounds were suitable for negative ion mode detection due to mass spectrum signals of their dominant adduct stronger in negative ion mode than positive ion mode. [Conclusion] Based on the accurate molecular weight, the established identifying method can identify the 18 cotton metabolite standards with their nontarget mass spectrometry data. Under the determined UPLC and ESI condition, the dominant adducts of cotton secondary metabolites have the preference of ESI mode. These results provide technical and theoretical data support for further study of cotton metabolomics.

Key words: cotton metabolite; ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS); mass spectrum identification; adduct