中药饮片中真菌毒素前处理方法进展
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摘要
中药真菌毒素是真菌在中药中所产生的次生代谢产物,一般不表现出急性毒性,但具有较强的蓄积性。我国常用中药存在真菌毒素污染或潜在污染的情况。为了改善中药基质对真菌毒素检测结果的影响,提高中药真菌毒素检测灵敏度与准确度,保证人类健康,减少真菌毒素带来的经济损失,本文对现代中药真菌毒素提取和净化方法进行了总结和概括,主要包括高速均质提取、搅拌提取、超声提取、振荡提取及免疫亲和柱净化法、固相萃取柱净化法、QuEChERS净化法等,并对真菌毒素前处理方法进行展望,以供国内外同行参考借鉴。
Mycotoxins are secondary metabolites produced by certain fungi in traditional Chinese medicine( TCM),which generally don't show acute toxicity but have strong accumulation. Traditional Chinese medicines are contaminated or potentially polluted by mycotoxins. In order to improve the effect of traditional Chinese medicine matrix on mycotoxin detection,to improve the sensitivity and accuracy of mycotoxin detection,and to ensure human health and reduce the economic loss caused by mycotoxins,this paper summarizes the extraction and purification methods of mycotoxin from TCM which mainly include high-speed homogenization extraction,stirring extraction,ultrasonic extraction,oscillation extraction and immunoaffinity column( ICA) purification,solid phase extraction column( SPE) purification and Qu ECh ERS purification. Some expectation of the methods of mycotoxin pretreatment was brought forward in this article for providing reference for peers at home and abroad.
Mycotoxins are secondary metabolites produced by certain fungi in traditional Chinese medicine( TCM),which generally don't show acute toxicity but have strong accumulation. Traditional Chinese medicines are contaminated or potentially polluted by mycotoxins. In order to improve the effect of traditional Chinese medicine matrix on mycotoxin detection,to improve the sensitivity and accuracy of mycotoxin detection,and to ensure human health and reduce the economic loss caused by mycotoxins,this paper summarizes the extraction and purification methods of mycotoxin from TCM which mainly include high-speed homogenization extraction,stirring extraction,ultrasonic extraction,oscillation extraction and immunoaffinity column( ICA) purification,solid phase extraction column( SPE) purification and Qu ECh ERS purification. Some expectation of the methods of mycotoxin pretreatment was brought forward in this article for providing reference for peers at home and abroad.
引文
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[24]李梦华.中药及其相关产品中多种真菌毒素同步检测研究[D].镇江:江苏大学,2016.
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[32]YangYing,WenJing,KongWeijun,et al.Simultaneous Determination of Four Aflatoxins and Ochratoxin A in Ginger after Inoculation with Fungi by Ultra-Fast Liquid Chromatography-Tandem Mass Spectrometry[J].Journal of the Science of Food and Agriculture,2016,96(12):4160-4167.
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[34]Wen J,Kong W,Wang J,et al.Simultaneous Determination of Four Aflatoxins and Ochratoxin A In Ginger and Related Products by HPLC with Fluorescence Detection after Immunoaffinity Column Clean-Up and Postcolumn Photochemical Derivatization[J].Journal of Separation Science,2013,36(23):3709-16.
[35]李春,苏晓,冯伟红,等.基于UPLC-ESI-MS/MS的何首乌中12种真菌毒素污染检测[J].中国中药杂志,2016,41(23):4368-4374.
[36]王少敏,许勇,毛丹,等.HPLC-MS/MS法测定中药桃仁中黄曲霉毒素G_2、G_1、B_2、B_1[J].药物分析杂志,2011,31(5):907-911.
[37]魏俊德,邱海蕴,慎洁.高效液相色谱-串联质谱法测定白苏子中黄曲霉毒素[C]//国际数字医学会数字中医药分会成立大会暨首届数字中医药学术交流会论文集.2016,131.
[38]郑荣,王少敏,简龙海,等.薏苡仁中7种真菌毒素的液相色谱-串联质谱测定法[J].中国卫生检验杂志,2011,21(2):318-320.
[39]郑荣,毛丹,王柯,等.HPLC法测定中药中黄曲霉毒素B1、B2、G1、G2的含量[J].药物分析杂志,2005,25(6):610-613.
[40]郑荣,毛丹,王柯,等.柱后衍生-高效液相色谱法测定酸枣仁中黄曲霉毒素G2、G1、B2、B1[J].中国卫生检验杂志,2010,20(1):36-37.
[41]杨文武,熊凌云,王瑞芳,等.免疫亲和柱净化HPLC挂后光化学衍生法测定34批中药材中黄曲霉毒素G2、G1、B2、B1[J].现代中药研究与实践,2013,27(1):43-47.
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[47]李峻媛.薏苡仁中常见真菌毒素的检测方法及不同储藏条件下真菌毒素含量变化的研究[D].成都:成都中医药大学,2012.
[2]辽宁出入境检验检疫局网站.关于韩国制订中药材中二氧化硫和黄曲霉毒素相关标准事宜的通知[EB/OL].http://www.lnciq.gov.cn/ywpd/zwjy/ywbl/czjy/200804/t20080416_4539.htm,2008-04-16/2010-04-01.
[3]WM/T2-2004,药用植物及制剂进出口绿色行业标准[S].中华人民共和国商务部,2004,2.
[4]国家药典委员会.中华人民共和国药典附录I[S].北京:中国医药科技出版社,2010:附录I:402-404.
[5]傅勇,曹纪亮,杨小丽,等.真菌毒素检测的前处理技术及其应用[J].中国卫生检验杂志,2012,22(6):1462-1466.
[6]韩铮.中药材中常见真菌毒素分析方法学及代谢动力学研究[D].杭州:浙江大学,2011.
[7]Katerere D R,Stockenstr9m S,Thembo K M,et al.A Preliminary Survey of Mycological and Fumonisin and Aflatoxin Contamination of African Traditional Herbal Medicines Sold in South Africa[J].Human&Experimental Toxicology,2008,27(11):793.
[8]Zheng H,Zheng Y,Luan L,et al.An Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Simultaneous Determination of Aflatoxins B1,B2,G1,G2,M1 And M2 in Traditional Chinese Medicines[J].Analytica Chimica Acta,2010,664(664):165-171.
[9]Han Z,Liu X,Ren Y,et al.A Rapid Method with Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry for Simultaneous Determination of Five Type B Trichothecenes in Traditional Chinese Medicines[J].Journal of Separation Science,2010,33(13):1923-32.
[10]Han Z,Ren Y,Liu X,et al.A Reliable Isotope Dilution Method for Simultaneous Determination of Fumonisins B1,B2 And B3 in Traditional Chinese Medicines by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry[J].Journal of Separation Science,2010,33(17-18):2723-2733.
[11]Zhang L,Dou X,Kong W,et al.Assessment of Critical Points and Development of A Practical Strategy to Extend the Applicable Scope of Immunoaffinity Column Cleanup for Aflatoxin Detection in Medicinal Herbs[J].Journal of Chromatography A,2016,12(1483):56.
[12]Arroyo-Manzanares N,García-Campa1a A M,Gámiz-Gracia L.Multiclass Mycotoxin Analysis in Silybummarianum by Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry Using A Procedure Based on Quechers and Dispersive Liquid-Liquid Microextraction[J].Journal of Chromatography A,2013,1282(5):11.
[13]李鹏,贺澎,欧国静.中药黄曲霉毒素检测方法学的改良研究[J].中国医疗前沿,2012,7(6):71-71.
[14]孔祥虹,高军刚,何强,等.超高效液相色谱-串联质谱法测定植物提取物中的4种黄曲霉毒素[J].食品科技,2013,38(4):325-329.
[15]Zheng R,Xu H,Wang W,et al.Simultaneous Determination of Aflatoxin B(1),B(2),G(1),G(2),Ochratoxin A,and Sterigmatocystin in Traditional Chinese Medicines by LC-MS-MS[J].Analytical and Bioanalytical Chemistry,2014,406(13):3031.
[16]林方芬,郑志勇,黄剑,等.中药饮片中黄曲霉毒素B1含量的快速测定研究[J].江苏中医药,2016,48(12):65-66,69.
[17]Zheng R S,Wang W L,Tan J,et al.An Investigation of Fungal Contamination on The Surface of Medicinal Herbs in China[J].Chinese Medicine,2017,12(1):2.
[18]Ali N,Hashim N H,Saad B,et al.Evaluation of A Method to Determine the Natural Occurrence of Aflatoxins in Commercial Traditional Herbal Medicines from Malaysia and Indonesia[J].Food&Chemical Toxicology,2005,43(12):1763-1772.
[19]付成平,欧阳华学,刘瑜.柱前衍生化高效液相色谱法测定中草药提取物中的黄曲霉毒素B1[J].西南农业学报,2009,22(2):522-524.
[20]陈重均.超高效液相色谱串联质谱定量分析中药材及其制剂中真菌毒素方法的研究[D].杭州:浙江大学,2015.
[21]李峻媛.薏苡仁中常见真菌毒素的检测方法及不同储藏条件下真菌毒素含量变化的研究[D].成都:成都中医药大学,2012.
[22]胡一晨,万丽,范成杰,等.免疫亲和柱净化HPLC柱后光化学衍生化法检测中药及染菌中药制剂中间体的黄曲霉毒素[J].中国实验方剂学杂志,2012,18(10):116-119.
[23]赵连华.陈皮中黄曲霉毒素的累积对其质量影响的研究[D].长春:吉林农业大学,2015.
[24]李梦华.中药及其相关产品中多种真菌毒素同步检测研究[D].镇江:江苏大学,2016.
[25]胡一晨.中药及其提取物中农药多残留同步分析方法及黄曲霉毒素净化方法的研究[D].成都:成都中医药大学,2012.
[26]朱迪,谭丹,向文英,等.免疫亲和柱-HPLC柱后光化学衍生法测定中药饮片中黄曲霉毒素B1、B2、G1、G2的含量[J].贵阳医学院学报,2015,40(8):843-847.
[27]Kong W J,Li J Y,Qiu F,et al.Development of A Sensitive and Reliable High Performance Liquid Chromatography Method with Fluorescence Detection for High-Throughput Analysis of Multi-Class Mycotoxins in Coixseed[J].Analytica Chimica Acta,2013,799(17):68-76.
[28]Yang L,Wang L N,Pan J Y,et al.Determination of Ochratoxin A in Traditional Chinese Medicinal Plants by HPLC-FLD[J].Food Additives&Contaminants Part A Chemistry Analysis Control Exposure&Risk Assessment,2010,27(7):989-997.
[29]郝爱鱼,赵丽元,刘英慧,等.HPLC柱后光衍生荧光法测定中药饮片中黄曲霉毒素残留量[J].药物分析杂志,2012(12):2203-2207.
[30]Wei R,Qiu F,Kong W,et al.Co-Occurrence of Aflatoxin B 1,B 2,G 1,G 2,and Ochrotoxin A In Glycyrrhizauralensis,Analyzed By HPLC-MS/MS[J].Food Control,2013,32(1):216-221.
[31]曹纪亮.中药鸦胆子和生姜中黄曲霉毒素和赭曲霉毒素A的检测研究[D].成都:成都中医药大学,2013.
[32]YangYing,WenJing,KongWeijun,et al.Simultaneous Determination of Four Aflatoxins and Ochratoxin A in Ginger after Inoculation with Fungi by Ultra-Fast Liquid Chromatography-Tandem Mass Spectrometry[J].Journal of the Science of Food and Agriculture,2016,96(12):4160-4167.
[33]韦日伟,杨小丽,仇峰,等.免疫亲和柱净化-在线柱后光化学衍生-HPLC-FLD同时测定甘草中黄曲霉毒素B1,B2,G1,G2和赭曲霉毒素A的含量[J].中国中药杂志,2011,36(17):2342-2346.
[34]Wen J,Kong W,Wang J,et al.Simultaneous Determination of Four Aflatoxins and Ochratoxin A In Ginger and Related Products by HPLC with Fluorescence Detection after Immunoaffinity Column Clean-Up and Postcolumn Photochemical Derivatization[J].Journal of Separation Science,2013,36(23):3709-16.
[35]李春,苏晓,冯伟红,等.基于UPLC-ESI-MS/MS的何首乌中12种真菌毒素污染检测[J].中国中药杂志,2016,41(23):4368-4374.
[36]王少敏,许勇,毛丹,等.HPLC-MS/MS法测定中药桃仁中黄曲霉毒素G_2、G_1、B_2、B_1[J].药物分析杂志,2011,31(5):907-911.
[37]魏俊德,邱海蕴,慎洁.高效液相色谱-串联质谱法测定白苏子中黄曲霉毒素[C]//国际数字医学会数字中医药分会成立大会暨首届数字中医药学术交流会论文集.2016,131.
[38]郑荣,王少敏,简龙海,等.薏苡仁中7种真菌毒素的液相色谱-串联质谱测定法[J].中国卫生检验杂志,2011,21(2):318-320.
[39]郑荣,毛丹,王柯,等.HPLC法测定中药中黄曲霉毒素B1、B2、G1、G2的含量[J].药物分析杂志,2005,25(6):610-613.
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