UPLC/MS/MS法定性定量测定中药材中3类27种合成色素(1)
摘要 目的：采用液相-固相萃取-超高效液相色谱-电喷雾串联质谱联用技术对27种色素建立了准确、灵敏的分析方法。方法：针对脂溶性、酸性和碱性3类色素分别采用不同的制备方法、色谱条件和电喷雾极性，采用4种不同药用部位的代表性药材进行了方法学考察。结果：表明方法线性关系良好(r≥0.997);脂溶性和碱性色素的检测限为0.03～3.87 ng/g，酸性色素为0.003～1.94 μg/g;准确度和精密度分别为70%～105%和0.9%～6.8%;应用于25个品种80批样品的筛查。结论：所建方法可靠、高效、准确，适用于中药材复杂基质中不同种类色素的测定。
关键词 27种;合成色素;脂溶性;酸性;碱性;定性定量;中药材;UPLC/MS/MS
Qualitative and Quantitative Determination of 27 Synthetic Dyes of 3 Species in Herbs by UPLC/MS/MS
Zhang Su1，2，Hu Qing1，Sun Jian1，Feng Rui1，Yu Hong1，Ji Shen1
(Shanghai Institute for Food and Drug Control，Shanghai 201203，China)
Abstract Objective：To develop a sensitive and accurate method combining liquid and solid phase extraction with ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry to analyze 27 dyes.Methods：Different preparations，chromatography conditions and polarity of ESI were applied for 3 categories of lipophilic，acidic and basic dyes respectively.Four species of herbs representing different parts of plant were selected for methodology observation.Results：The linear correlation coefficient was good(r ≥ 0.997).The LODs of oil soluble and basic dyes were 0.03～3.87 ng/g and acid dyes were 0.003～1.94 μg/g respectively.The accuracies and precisions were 70%～105% and 0.9%～6.8% respectively.Conclusion：After detecting 80 samples of 25 species herbs，this method proved to be reliable，efficient and accurate in the detection of different kinds of dyes in the complex matrix of herbs.
Key Words 27 kinds; Artificial dyes; Lipophilic，Acidity; Basicity; Qualitative and quantitative determination; Herbs; UPLC/MS/MS
中圖分类号：R284.1文献标识码：Adoi：10.3969/j.issn.1673-7202.2019.04.007
中药主要为天然产物，随着社会发展，人们对身体健康愈加重视，市场对中药的需求不断增加，价格也水涨船高。由于当前中药生产、经营秩序尚未完全规范，社会诚信体系缺失，监管无法涉及所有环节，中药染色、以劣充好等违法违规行为仍屡禁不止[1-3]。这些违法行为严重危害人民身体健康和生命安全、影响中医中药的声誉和中医药事业的健康发展。特别是合成色素的添加，其特征的偶氮基团(-N=N-)结构，易被肠道菌群还原形成芳香胺类化合物[4]，可引起神经毒性[5]、基因毒性[6]以及致癌性[7]。
目前，合成色素的分析方法包括薄层色谱法(TLC)[8]、毛细管电泳法(CE)[9]、离子色谱法[10]、高效液相色谱法(HPLC)[11-12]、液相色谱-串联质谱法(LC-MS/MS)[13-14]、液相色谱-四极杆飞行时间质谱法[15-16]等。本实验室已建立了快速检测的薄层色谱法，粗略定量和定性的高效液相色谱法[17]以及用于定性的UHPLC-Q-TOF法[18]。这些方法可在不同水平的实验室用于不同处理过程的色素测定。本文重点阐述利用UPLC/MS/MS对色素同时进行定量和定性分析。对3组色素的流动相和质谱条件进行优化，为体现不同基质的复杂性，选用4种不同药用部位的代表性药材进行了方法学考察。应用该方法对25个品种80批样品进行了测定，结果表明该方法灵敏、准确，适用于中药材基质中染色色素的测定。
1 仪器与试药
1.1 仪器 采用超高效液相色谱-串联质谱联用技术(UPLC/MS/MS)，在正、负离子2种采集模式下，对复杂中药基质中的目标化合物进行了鉴定。Waters ACQUITY UPLC I-Class(Milford，Massachusetts，USA)由二元泵(BSM)、进样器(SM-FTN)和柱温箱(CM-A)组成，配有AB Sciex API 4000 MS/MS系统(Framingham，Massachusetts，USA)，采用Analyst 1.6.1软件进行数据采集和处理。, http://www.100md.com(张甦 胡青 孙健 冯睿 于泓 季申)
关键词 27种;合成色素;脂溶性;酸性;碱性;定性定量;中药材;UPLC/MS/MS
Qualitative and Quantitative Determination of 27 Synthetic Dyes of 3 Species in Herbs by UPLC/MS/MS
Zhang Su1，2，Hu Qing1，Sun Jian1，Feng Rui1，Yu Hong1，Ji Shen1
(Shanghai Institute for Food and Drug Control，Shanghai 201203，China)
Abstract Objective：To develop a sensitive and accurate method combining liquid and solid phase extraction with ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry to analyze 27 dyes.Methods：Different preparations，chromatography conditions and polarity of ESI were applied for 3 categories of lipophilic，acidic and basic dyes respectively.Four species of herbs representing different parts of plant were selected for methodology observation.Results：The linear correlation coefficient was good(r ≥ 0.997).The LODs of oil soluble and basic dyes were 0.03～3.87 ng/g and acid dyes were 0.003～1.94 μg/g respectively.The accuracies and precisions were 70%～105% and 0.9%～6.8% respectively.Conclusion：After detecting 80 samples of 25 species herbs，this method proved to be reliable，efficient and accurate in the detection of different kinds of dyes in the complex matrix of herbs.
Key Words 27 kinds; Artificial dyes; Lipophilic，Acidity; Basicity; Qualitative and quantitative determination; Herbs; UPLC/MS/MS
中圖分类号：R284.1文献标识码：Adoi：10.3969/j.issn.1673-7202.2019.04.007
中药主要为天然产物，随着社会发展，人们对身体健康愈加重视，市场对中药的需求不断增加，价格也水涨船高。由于当前中药生产、经营秩序尚未完全规范，社会诚信体系缺失，监管无法涉及所有环节，中药染色、以劣充好等违法违规行为仍屡禁不止[1-3]。这些违法行为严重危害人民身体健康和生命安全、影响中医中药的声誉和中医药事业的健康发展。特别是合成色素的添加，其特征的偶氮基团(-N=N-)结构，易被肠道菌群还原形成芳香胺类化合物[4]，可引起神经毒性[5]、基因毒性[6]以及致癌性[7]。
目前，合成色素的分析方法包括薄层色谱法(TLC)[8]、毛细管电泳法(CE)[9]、离子色谱法[10]、高效液相色谱法(HPLC)[11-12]、液相色谱-串联质谱法(LC-MS/MS)[13-14]、液相色谱-四极杆飞行时间质谱法[15-16]等。本实验室已建立了快速检测的薄层色谱法，粗略定量和定性的高效液相色谱法[17]以及用于定性的UHPLC-Q-TOF法[18]。这些方法可在不同水平的实验室用于不同处理过程的色素测定。本文重点阐述利用UPLC/MS/MS对色素同时进行定量和定性分析。对3组色素的流动相和质谱条件进行优化，为体现不同基质的复杂性，选用4种不同药用部位的代表性药材进行了方法学考察。应用该方法对25个品种80批样品进行了测定，结果表明该方法灵敏、准确，适用于中药材基质中染色色素的测定。
1 仪器与试药
1.1 仪器 采用超高效液相色谱-串联质谱联用技术(UPLC/MS/MS)，在正、负离子2种采集模式下，对复杂中药基质中的目标化合物进行了鉴定。Waters ACQUITY UPLC I-Class(Milford，Massachusetts，USA)由二元泵(BSM)、进样器(SM-FTN)和柱温箱(CM-A)组成，配有AB Sciex API 4000 MS/MS系统(Framingham，Massachusetts，USA)，采用Analyst 1.6.1软件进行数据采集和处理。, http://www.100md.com(张甦 胡青 孙健 冯睿 于泓 季申)