免疫亲和柱净化-在线柱后光化学衍生HPLC-FLD检测莲子中黄曲霉毒素B_1,B_2,G_1,G_2及其液质确证  

Determination of aflatoxins in Nelumbinis Semen by immunoaffinity column clean-up and HPLC-FLD with on-line post-column photochemical derivatization and LC-MS/MS confirmation

作　　者：刘书宇[1] 仇峰[1] 杨美华[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植研究所中草药物质基础与资源利用教育部重点实验室,北京100094

出　　处：《中国中药杂志》2012年第3期305-309,共5页China Journal of Chinese Materia Medica

基　　金：国家"重大新药创制"科技重大专项(2009ZX09502-025;2010ZX09301-002-001);国家中医药管理局中医药行业科研专项(200807042)

摘　　要：目的:建立高效液相色谱-在线柱后光化学衍生-荧光检测器检测药食同源中药材莲子中黄曲霉毒素B1,B2,G1,G2的含量,并采用液质联用法进行确证。方法:样品以甲醇-水(80∶20)溶液提取,经免疫亲和柱净化后,利用在线柱后光化学衍生-HPLC-FLD进行分析测定。并进一步采用LC-MS/MS对阳性样品进行确证。结果:在优化条件下,黄曲霉毒素B1,G1在0.3～30μg.L-1,黄曲霉毒素B2,G2在0.09～9.0μg.L-1线性关系良好,r>0.999 9,回收率86.7%～99.1%,RSD<4.87%。黄曲霉毒素B1,B2,G1,G2的检测限(LOD)分别为0.08,0.03,0.10,0.03μg.kg-1。所测的20批莲子样品中,有14批样品的黄曲霉毒素检测结果呈阳性,其中黄曲霉毒素B1的污染水平为0.40～586μg.kg-1,黄曲霉毒素总量(B1+B2+G1+G2)的污染水平为0.40～602.5μg.kg-1。通过LC-MS/MS确证,在与对照品相同的保留时间处,样品与对照品有相同的特征离子碎片,排除了样品假阳性的可能。结论:该方法简便快速,灵敏度高,重复性好,适用于莲子中黄曲霉毒素的检测。To determine the contents of aflatoxin B1, B2, G1 and G2 in Nelumbinis Semen using on-line post-column photo- chemical derivatization-HPLC-FLD method and verify the method by LC-MS method. Method： The samples were extracted with MeOH-H20 （80： 20） and purified with inmunoaffinity column, aflatoxins were analyzed by HPLC-FLD with post-column photochemical derivatizaton. The positive samples were further confirmed by LC-MS/MS. Result： On optimum conditions, aflatoxin B1, G1 ranging 0. 3-30 mg·L-1 showed a good linear relationship with aflatoxin B2 , G2 ranging 0. 09-9. 0 mg ·L- 1 with r 〉 0. 999 9. The recoveries ranged between 86. 7% and 99. 1% , with RSDs all bellow 4. 87%. LOD of aflatoxin B1 , B2 , GI and G： were 0. 08, 0. 03, 0. 10, 0. 03 μg·kg^-1 , respectively. Among 20 Nelumbinis Semen samples, 14 were found to contain aflatoxin B1 ranging from 0. 40 to 586μg·kg^-1. The total content of aflatoxin B1 , B2 , G1 and G2 were between 0. 40 and 602. 5μg·kg^-1. By LC-MS/MS method, the same fragment ions were founded in samples and the control group at the same retention times, ruling out the possibility of false positive samples. Conclusion： The method is simple, highly sensitive and reproducible for the determination of aflatoxins in Nelumbinis SeInOFI.

关 键 词：黄曲霉毒素 莲子 在线光化学衍生 高效液相色谱 液质确证 

分 类 号：R284.1[医药卫生—中药学]