高效液相色谱-四极杆-飞行时间质谱快速筛查鉴别食品中非法添加的62种中药材北大核心CSCD

建立了高效液相色谱-四极杆-飞行时间质谱快速筛查鉴别食品中非法添加的62种中药材的方法。依据卫生部关于进一步规范保健食品原料管理的通知(卫法监发[2002]51号)中可用于保健食品的物品名单,确定食品中可能非法添加的62种中药材原料清单,再从62种中药材中筛选其特征组分,获得不同中药材对应的特征组分清单。62种对照药材经甲醇超声提取后,于Thermo Accucore aQ色谱柱(150 mm×2.1 mm,2.6μm)上分离,在电喷雾正负离子扫描模式下,分别以0.1%(v/v)甲酸水溶液-乙腈和水-乙腈为流动相梯度洗脱,进行一级质谱和二级质谱全扫描检测,采用Library View软件建立不同中药材对应的特征组分的一级精确质量数据库和二级碎片质谱库。样品同法处理后上样分析,采用Peak View软件将样品高分辨数据与自建数据库中的质谱图、精确分子离子质量数、碎片离子质量数、保留时间等相关参数进行快速筛查鉴别分析。该工作通过建立“中药材-特征组分”对应清单,构建了食品中易非法添加的62种中药材中共388种特征组分的高分辨质谱库,每种中药材包括5~10种特征组分,通过对实际食品样品配制酒、代用茶及饮料进行筛查分析,1批次配制酒样品与淫羊藿中药材的7种特征组分匹配一致,推断该配制酒样品中加入了淫羊藿中药材。该法可实现无标准品情况下中药材的定性筛查,具有高通量、准确、简便、快捷的特点,解决了食品中非法添加中药材难以识别和确证的难题,可以实现食品中非法添加中药材的快速筛查鉴别分析。

Rapid screening and identification of 62 kinds of illegally added traditional Chinese medicine in food by high performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry

An analytical method based on high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(HPLC-Q-TOF/MS)was established for the rapid screening and identification of 62 kinds of illegally added traditional Chinese medicine(TCM)in food.According to the notice of the Ministry of Health of the People’s Republic of China on further regulating the management of raw materials of health food(Weifa Jianfa(2002)No.51),the characteristic components of the 62 kinds of TCM were screened,and the corresponding characteristic component lists of different TCM were obtained.Methanolic extracts of the 62 kinds of standard medicinal materials were subjected to HPLC-Q-TOF-MS analysis.The filtrate was separated on a Thermo Accucore aQ column(150 mm×2.1 mm,2.6μm)using 0.1%(v/v)formic acid aqueous solution or water and acetonitrile as the mobile phases for gradient elution in the electrospray positive and negative ion scanning mode.All the data were determined on the full scan of primary mass spectrometry and secondary mass spectrometry,with mass acquisition ranges of 100-1000 Da and 50-1000 Da,respectively.A 10 mmol/L sodium formate solution was used as the mass correction solution in both the positive and negative ion modes.Library View software was used to establish the precursor ion accurate quality database and the product ion fragment mass spectrometry database of the corresponding characteristic components of the different kinds of TCM.In the Library View database software,the name of each characteristic component of the 62 kinds of TCM was input(serial number)in order to classify the screened characteristic components.The samples were processed using the same method and analyzed.Peak View software was used to rapidly analyze and screen the target components of the TCM.The high-resolution data collected from the samples to be tested were imported into the Peak View software,followed by the compound list of the established MS database of standard medicinal materials.After setting the identification method parameters and library retrieval parameters,a matching analysis was performed,and the candidate substances for each peak were automatically identified by comparing the mass spectrum,accurate molecular ion mass number,fragment ion mass number,retention time,and other related parameters.The determination conditions of compound detection were as follows:the comprehensive score was more than 70 points.The molecular formula,retention time,mass spectrum as well as the primary isotope mass spectrometry,primary mass spectrometry,and secondary mass spectrometry data were matched with the library compounds.The corresponding list of“TCM-characteristic components”was established,and a high-resolution MS library of 388 characteristic components from the 62 types of TCM was constructed.Each TCM contains 5-10 characteristic components.According to the screening analysis of the actual food samples of the prepared wine,substitute tea,and beverage,one batch of the prepared wine sample matched with seven characteristic components of epimedium,and it was inferred that epimedium was added to the prepared wine samples.This method can allow for the qualitative screening of TCM without standards and has the characteristics of high throughput,accuracy,simplicity,and rapidity.It solves the difficulty in identifying and confirming illegally added TCM in food;provides technical methods and a basis for cracking down on the illegal addition of TCM in food;and facilitates the rapid screening and identification of illegally added TCM in food.