학술논문
UPLC-MS/MS法与ELISA法测定胖大海中黄曲霉毒素的比较研究 / Comparative study on determination of aflatoxins in Sterculiae Lychnophorae Semen by UPLC-MS/MS and ELISA
Document Type
Academic Journal
Author
Source
中国药师 / China Pharmacist. 26(10):168-175
Subject
Language
Chinese
ISSN
1008-049X
Abstract
目的 采用超高效液相色谱-串联质谱法(UPLC-MS/MS)和酶联免疫吸附法(ELISA)对胖大海中的黄曲霉毒素进行检测,比较两种方法的检测结果以及方法学差异性.方法 UPLC-MS/MS法测定:样品采用 70%甲醇提取经过免疫亲和柱净化,色谱柱为C18(100 mm×2.1 mm,1.7 μm),柱温为40℃,流动相为水含0.1%甲酸(A)-甲醇含0.1%甲酸(B),梯度洗脱,流速为0.3 mL·min-1,多反应检测扫描采集模式,正负离子同时扫描进行数据采集.同时采用ELISA法测定,样品采用甲醇提取,酶联免疫试剂盒反应.结果 UPLC-MS/MS法测定:黄曲霉毒素B1、B2、G1、G2 线性关系良好,检测限分别为 0.0135,0.0029,0.0086,0.0036 μg·kg-1,定量限分别为 0.0450,0.0098,0.0286,0.0119 μg·kg-1,平均回收率分别为 98.98%,100.32%,98.53%,99.56%,RSD≤2.42%(n=9).ELISA法测定:黄曲霉毒素B1 和总量的定量限分别为0.16,0.08 μg·kg-1,检出限分别为 0.05,0.02 μg·kg-1,平均回收率分别为 93.07%,89.67%,RSD≤13.99%(n=9).两种方法测定结果的标准偏差在 3.46%～7.63%之间.结论 UPLC-MS/MS法的定量限低,检测限较低,加样回收率高,灵敏度和准确度较高,结果可靠.与UPLC-MS/MS法相比,ELISA法测定胖大海中黄曲霉毒素总量和黄曲霉毒素B1 结果差异较小,未出现假阳性等情况.