液质联用法快速筛查中药材及饮片中常见染色掺假物质的方法研究及数据库建立

目的：建立基于液质联用法的中药材及饮片中常见染色掺假物质的快速筛查方法。方法：采用Poroshell 120 EC-C₁₈ 色谱柱（3.0 mm×100 mm，2.7 μm），以乙腈和0.02 mol·L^-1 的乙酸铵水溶液为流动相进行梯度洗脱，流速0.4 mL·min^-1，柱温35 ℃，检测波长为428、483、509、608 nm；采用电喷雾电离源（ESI），正离子模式检测，扫描质谱扫描范围m/z 100~1 000。将胭脂红、偶氮玉红、苋菜红、赤藓红、罗丹明B、酸性红73、诱惑红、中性红、刚果红、甲基红、曙红、新品红、金光红C、橙G、碱性橙Ⅱ、金胺O、金橙Ⅱ、柠檬黄、日落黄、酸性橙Ⅰ、皂黄、碱性橙21、碱性橙22、孔雀石绿、亮绿、亮蓝、结晶紫、专利蓝、亚甲基蓝、苏丹红Ⅰ、苏丹红Ⅱ、苏丹红Ⅲ、苏丹红Ⅳ、苏丹红G、分散红9、808 猩红、柑橘红2 及松香酸38 种已有或可能的染色掺假物质分别依次进行LC-MS/MS（QTOF）分析，结合MassHunter PCDL 软件建立数据库，以通过软件自动筛选出可疑染色掺假物质。结果：采用所建立的方法快速筛查验证了红花、西红花、血竭、朱砂、延胡索、黄连、蒲黄、五味子、熟地、制首乌、制黄精、全蝎、松花粉、山茱萸、乳香、没药、沉香、乌梅、海金沙、青黛20 种中药材及饮片共200 余批，对于各种药材及饮片基本实现通用，有效识别了补充检验方法未收载的染色掺假物质，验证了快速筛查方法的可行性。结论：建立的常见染色掺假物质数据库和分析方法具有快速、灵敏、准确的优点，节省了时间和实验成本。

Objective: To establish a rapid LC-MS/MS method for screening commonly used dyes in traditional Chinese medicinal materials and decoction pieces. Methods: Poroshell 120 EC-C₁₈ chromatographic column (3.0 mm×100 mm,2.7 μm)was used and the mobile phase was acetonitrile and 0.2 mol·L^-1 ammonium acetate solution with gradient elution at the flow rate of 0.4 mL·min^-1. The column temperature was 35℃ and the detection wavelengths were 428 nm,483 nm,509 nm and 608 nm. ESI source was operated in negative ion mode and the scan range was m/z 100-1 000. The purpose is to screen out suspicious dyeing adulterated material automatically by the LC-MS/MS(QTOF)data of 38 known or potential adulterated material(ponceau 4r,azorubin,amaranth, erythrosin B,rhodamine B,acid red 73,allura red,neutral red,congo red,methyl red,eosin Y,new fuchsin, lake red C,orange g,chrysoidine G,auramine O,orange Ⅱ,tartrazine,food yellow 3,orange I,metanil yellow, basic orange 21,basic orange 22,malachite green,brilliant green,brilliant blue FCF,basic violet 3,patent blue V,methylene blue,sudan Ⅰ,sudan Ⅱ,sudan Ⅲ,sudan Ⅳ,sudan red G,disperse red 9,808 scarlet,citrus red 2,abietic acid)acquired by using Agilent LC-MS optimized analysis condition and MassHunter PCDL software. Results: 20 traditional Chinese medicine materials(Carthami Flos,Croci Stigma,Draconis Sanguis,Cinnabar, Corydalis Rhizoma,Coptidis Rhizoma,Typhae Pollen,Schisandrae Chinensis Fructus,Rehmanniae Radix Praeparata,Polygoni Multiflori Radix Praeparata,Polygonati Rhizoma Praeparata,Scorpio,Pini Pollen,Corni Fructus,Olibanum,Myrrha,Aquilariae Lignum Resinatum,Mume Fructus,Lygodii Spora,Indigo Naturalis) with various colors for more 200 batches sample were successfully screened and identified. This method can be applied to different materials and can efficiently detect dyeing adulterated samples which have not been included in supplementary testing method,proved the practicability of this fast screening method. Conclusion: The established common dyeing adulterated material database and analysis method are proved to be fast,sensitive,and accurate. It also saves time and experimental cost.

[1] 高珊,张婷婷,崔邵婧,等. 市售蒲黄饮片的品质考察[J]. 中国药 事,2014,28(3):292 GAO S,ZHANG TT,CUI SJ,et al. Quality investigation on commercial Pollan Typhae decoction pieces[J]. Chin Pharm Aff, 2014,28(3):292
[2] 付凌燕,闵春艳,汪祺,等. 市售西红花药材掺伪染色检测方法的 实验研究[J]. 药物分析杂志,2012,32(1):74 FU LY,MIN CY,WANG Q,et al. Experimental research on the detection methods of Croci Stigma adulteration in the markets[J]. Chin J Pharm Anal,2012,32(1):74
[3] 肖凌,侯俊杰,聂晶,等. 元胡止痛系列制剂中金胺O 的检测[J]. 药物分析杂志,2012,32(11):2008 XIAO L,HOU JJ,NIE J,et al. Detection of auramine O in Yuanhu Zhitong preparations[J]. Chin J Pharm Anal,2012,32(11):2008
[4] 楚亮,周娟,王颖,等. 伪品人工牛黄中染色物质的鉴定[J]. 中成 药,2011,33(9):1556 CHU L,ZHOU J,WANG Y,et al. Identification of chemical industry dyes in adulteration of Bovis Calculus Artifactus[J]. Chin Tradit Pat Med,2011,33(9):1556
[5] 肖凌,侯俊杰,徐箐,等. 红花中一种新掺伪染料的确认及6 种染 色成分检测方法研究[J]. 药物分析杂志,2014,34(7):1231 XIAO L,HOU JJ,XU J,et al. Identification of a new adulteration dye and development of detection methods for six adulteration dyes in Carthami Flos[J]. Chin J Pharm Anal 2014,34(7):1231
[6] 翟清,杨志伟,石鑫磊,等. 染色蒲黄中柠檬黄的检测方法研究[J]. 中国医药科学,2015,5(19):69 ZHAI Q,YANG ZW,SHI XL,et al. Detection method of tartrazine in staining Typhae Pollen[J]. China Med Pharm,2015,5(19):69
[7] 饶伟文,蒋玲,赵纯玉,等. 几种染色掺假中药的化工染料鉴定[J]. 药物分析杂志,2007,27(11):1742 RAO WW,JIANG L,ZHAO CY,et al. Identification of chemical industry dyes in adulteration of Chinese medicinal materials[J]. Chin J Pharm Anal,2007,27(11):1742
[8] 姜国萍,孙萍,朱日然,等. 染色中药添加色素的检测技术研究进 展[J]. 时珍国医国药,2015,26(3):694 JIANG GP,SUN P,ZHU RR,et al. The dyeing testing technology research progress of Chinese medicine added pigment[J]. Lishizhen Med Mater Med Res,2015,26(3):694
[9] 薛虎寅,尹永梅,张太昌,等. 偶氮类合成色素检测技术的研究进 展[J]. 生物技术进展,2012,2(3):171 XUE HY,YIN YM,ZHANG TC,et al. Research on analysis of azo dyes in food products[J]. Curr Biotechnol,2012,2(3):171
[10] 李帮锐,冯家力,潘振球,等. 高效液相色谱-质谱/质谱联用法测定 饮料中的人工合成色素[J]. 中国卫生检验杂志,2007,17(4):579 LI BR,FENG JL,PAN ZQ,et al. Determination of synthetical pigments in drinks by high performance liquid chromatography-mass spectrometry/mass spectrometry[J]. Chin J Health Lab Technol, 2007,17(4):579
[11] 黄方取,曹丽军,严继东,等. 高效液相色谱-质谱/质谱联用法测 定葡萄酒中合成色素[J],中国卫生检验杂志,2013,23(15):3038 HUANG FQ,CAO LJ,YAN JD,et al. Determination of synthetical pigments in port wine by high performance liquid chromatographymass spectrometry/mass spectrometry[J]. Chin J Health Lab Technol,2013,23(15):3038
[12] 富小珺,张春梅. 几种染色药材中化工染料金胺O 的定性鉴别[J]. 中国医药指南,2013,11(21):107 FU XJ,ZHANG CM. The qualitative identification of auramine O in several stained traditional Chinese medicine[J]. Guide China Med, 2013,11(21):107
[13] 张智慧,阎正,蔡立鹏,等. HPCE 方法测定食品中皂黄色素[J]. 分析试验室,2009,28(S2):253 ZHANG ZH,YAN Z,CAI LP,et al. Determination of metanil yellow in food by HPCE[J]. Chin J Anal Lab,2009,28(S2):253
[14] 胡青,孙健,张甦,等. 中药中48 种非法染色色素的TLC 法检测[J]. 中国医药工业杂志2015,46(7):695 HU Q,SUN J,ZHANG S,et al. Simultaneous detection of 48 banned pigments in traditional Chinese medicine by TLC[J]. Chin J Pharm,2015,46(7):695
[15] 王学虎,袁敏,张前上,等. 液质联用仪自动筛选常见安眠镇静药 的应用研究[J]. 中国法医学杂志,2010,25(6):420 WANG XH,YUAN M,ZHANG QS,et al. A study on the method to screen hypnotics automaticaly by HPLC/MS[J]. Chin J Forensic Med,2010,25(6):420.
[16] 张燕飞,耿昭,黄再强,等. 朱砂中非法添加色素的HPLC-QTOF-MS 分析以及TLC 和HPLC 检测方法的建立[J]. 药物分 析杂志,2017(待发表) ZHANG YF,GENG Z,HUANG ZQ,et al. Qualitative analysis of illegally added pigment in cinnabar by HPLC-Q-TOF and establishment of TLC and HPLC analysis method[J]. Chin J Pharm Anal,2017(Article in Press)
[17] 耿昭,张燕飞,苟琰,等. 蒲黄中一种新染色掺假色素的确认及9 种橙黄色色素的检测方法研究[J]. 中国药房,2017(待发表) GENG Z,ZHANG YF,GOU Y,et al. Identification of a new adulterated dye and detection methods research for nine orangeyellow dyes in Typhae Pollen[J]. China Pharm(Article in Press)