超高效液相色谱法测定食用菌中L-麦角硫因及质谱确证

Determination of L-ergothioneine in edible fungi by ultra performance liquid chromatography and confirmation by mass spectrometry

建立了超高效液相色谱(UPLC)结合亲水相互作用色谱(HILIC)测定食用菌中L-麦角硫因含量的方法,并采用UPLC-四极杆串联飞行时间质谱(UPLC-Q-TOF-MS)进行确证。对食用菌样品前处理及色谱条件进行优化,样品经提取过膜后,采用HILIC-UPLC检测,外标法定量。结果显示,在不同食用菌样品中,加标回收率为80.0%~120.0%;日内、日间相对标准偏差(RSD)范围分别为0.4%~4.6%和1.7%~6.0%;采用科克伦和格拉布斯检验法,对7个不同实验室测定的4个相同食用菌样品数据的有效性进行检验,全部符合要求,计算得其重复性、再现性相对标准偏差范围分别为1.6%~2.4%和2.7%~7.5%。采用该方法对23个食用菌样品进行测定,其中牛肝菌中L-麦角硫因含量最高。方法可用于食用菌中L-麦角硫因含量的检测。

A method was developed for the determination of L-ergothioneine in edible fungi by ultraperformance liquid chromatography(UPLC)coupled hydrophilic interaction liquid chromatography(HILIC),and the confirmation analysis was achieved by UPLC-Q-TOF-MS.After optimizing the edible fungi sample pretreatment method and chromatographic conditions,the extracts were filtered through 0.22µm filters and analyzed by HILIC-UPLC.The external standard method was used for quantitation.The spiked recoveries of L-ergothioneine from the samples of edible fungiafter ranged from 80.0%to 120.0%.The relative standard deviations(RSDs)of intra-day and inter-day were 0.4%-4.6%and 1.7%-6.0%,respectively.Seven different laboratories were applied in this experiment to test the contents of L-ergothioneine in four kinds of edible fungi.Cochrane and Grubbs tests were used to test the validity of the testing data,and they all met the requirements.The RSDs of repeatability and reproducibility were calculated to be 1.6%-2.4%and 2.7%-7.5%,respectively.This method was used to determine 23 edible fungi samples,among which the L-ergothioneine content in boletus was the highest.The developed method is suitable for the determination and confirmation of L-ergothioneine content in edible fungi.