基于超高效液相色谱/高分辨飞行时间质谱正模式电喷雾下6个花色素裂解规律和特征离子研究  

Study on cleavage regularity and characteristic ions of six anthocyanins under ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry positive mode electric spray

作　　者：陈子凡 董莹 池少铃 郑少峰 罗凌超 王国胜 张南峰 CHEN Zi-fan;DONG Ying;CHI Shao-ling;ZHENG Shao-feng;LUO Ling-chao;WANG Guo-sheng;ZHANG Nan-feng(Heyuan Food Inspection Institute,Heyuan 517000,China;Urumqi Customs District,Dongguan 523000,China;Heyuan Drug Inspection Institute,Heyuan 517000,China;Institute of Zhejiang University-Quzhou,Quzhou 324000,China;Guangzhou Customs Technology Center Heyuan Branch,Heyuan 517000,China)

机构地区：[1]河源市食品检验所,河源517000 [2]黄埔海关技术中心,东莞523000 [3]河源市药品检验所,河源517000 [4]浙江大学衢州研究院,衢州324000 [5]广州海关技术中心河源分中心,河源517000

出　　处：《药物分析杂志》2024年第12期2051-2063,共13页Chinese Journal of Pharmaceutical Analysis

基　　金：河源市社会发展科技计划项目(230613111602379);广东省口岸安全智能化检测重点实验室开放课题资助项目(2023B1212010011);2017中央引导地方科技发展专项资金项目(2017245)。

摘　　要：目的:分析花葵素、矢车菊素、飞燕草素、芍药素、矮牵牛素和锦葵色素在超高效液相色谱/高分辨飞行时间质谱(UHPLC-Q TOF MS)正模式电喷雾下的裂解规律和途径,推导出6个花色素的特征产物离子,为建立质谱库以及检测方法提供理论依据。方法:采用Fusion-RPC_(18)(50 mm×2.0 mm,4μm)色谱柱,以0.1%甲酸水(A)-甲醇(B)为流动相,梯度洗脱(0~1 min,95%A;1~5 min,95%A→10%A;5~6 min,10%A;6~7 min,95%A),流速0.3 mL·min^(-1),柱温箱温度40℃,进样体积10μL。TOF MS-IDA MS/MS,CUR帘气0.20 MPa,碰撞气CAD为7 MPa,IS电压5500 V/-4500 V,离子源温度500℃,雾化气GAS1为0.38 MPa,辅助气GAS2为0.48 MPa,DP电压±60 V,裂解电压(35±15)V,累计时间0.2 s。在电喷雾正模式条件下,测得6个花色素的质谱数据,根据花色素的2-苯基色原烯结构形成的吡喃鎓离子,结合质谱仪数据库的辅助分析,推导出可能的产物离子。结果:6个花色素主要发生2种类型反应,一种是吡喃鎓环上发生裂解反应,最终均生成苯三酚和苄醇中间离子;另一种是发生α裂解以及σ裂解丢失环上的基团,最终生成不含基团的链烃。结论:该研究结果可以作为6个花色素的质谱特征离子特征数据的支撑,用于建立花色素产物离子库数据,也为检测方法开发研究提供参考。Objective:To analyze the fragmentation rule and pathway of pelargonidin,cyanidin,delphinidin,peonidin,petunidin and malvinidol under UHPLC-QTOF-MS positive mode electric spray,identify the characteristic product ions of six anthocyanins,and provide a theoretical basis for the establishment of mass spectrometry database and detection methods.Methods:The chromatographic conditions were as follows:chromatographic column,Fusion-RPC_(18)(50 mm×2.0 mm,4μm),mobile phase 0.1%formic acid aqueous solution(A)and methanol(B),gradient elution(0-1 min,95%A;1-5 min,95%A→10%A;5-6 min,1%A;6-7 min,95%A),flow rate 0.3 mL·min^(-1),column temperature 40℃,and injection volume 10μL.The mass spectrometry conditions were as follows:TOF MS-IDA MS/MS,curtain gas,0.20 MPa,collision gas CAD 7 MPa,IS voltage,5500 V/-4500 V,ion source temperature,500℃,nebulizer gas,GAS1,0.38 MPa,auxiliary,GAS2,0.48 MPa,DP voltage,±60 V,fragmentation voltage,(35±15)V,and time 0.2 s.Under the condition of positive mode of electrospray,the mass spectrometry data of six anthocyanins were measured.According to the pyrylium ions formed by the 2-phenylchromogenic structure of anthocyanins,and combined with the auxiliary analysis of the mass spectrometer database,the possible product ions were deduced.Results:The results showed that the six anthocyanins mainly undergo cleavage reactions on the pyranium ring,ultimately generating intermediate ions of pyrogallol and benzyl alcohol.On the other way,it occuredαcracking,σcracking causes the loss of functional groups on the ring,ultimately resulting in the formation of Chain hydrocarbons without functional groups.Conclusion:The research results can provide support for the mass spectrometry characteristic ion data of six anthocyanins,which can be used to establish ion library data for anthocyanin products and also provide reference for the development and research of detection methods.

关 键 词：花色素 正模式电喷雾 飞行质谱 裂解规律 特征离子 飞行时间质谱 

分 类 号：R917[医药卫生—药物分析学]