采用免疫亲和柱结合液相色谱-串联质谱法验证11 + Myco MS-PREP® 方法测定谷物、婴儿食品、香料和动物饲料中的黄曲霉毒素、伏马毒素、脱氧雪腐镰刀菌烯醇、赭曲霉毒素A、玉米赤霉烯酮、HT-2毒素和T-2毒素：AOAC性能验证方法SM 112401

Validation of the 11+Myco MS-PREP® Method for Determination of Aflatoxins, Fumonisins, Deoxynivalenol, Ochratoxin A, Zearalenone, HT-2, and T-2 Toxins in Cereals, Baby Food, Spices, and Animal Feed by Immunoaffinity Column with LC-MS/MS: AOAC Performance Tested MethodSM 112401.

作者信息

Leeman Dave, Allan Andrew B, Cameron Helen, Donelly Carol, Tramaseur Adam, Stratton Joanna, MacDonald Susan J

机构信息

R-Biopharm Rhône Ltd, Block 10 Todd Campus, West of Scotland Science Park, Acre Rd, Glasgow G20 0XA, UK.

Fera Science Ltd, Sand Hutton, York YO41 1LZ, UK.

出版信息

J AOAC Int. 2025 Mar 1;108(2):207-252. doi: 10.1093/jaoacint/qsae097.

DOI:10.1093/jaoacint/qsae097

PMID:39661480

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11879221/

Abstract

BACKGROUND

The 11+Myco MS-PREP® immunoaffinity column (IAC) contains a gel suspension of monoclonal antibodies specific to the toxins of interest. Following sample extraction, the IAC is used for cleanup and concentration of mycotoxins prior to analysis by LC with tandem mass spectrometry (LC-MS/MS).

OBJECTIVE

This study evaluated the IAC with LC-MS/MS method for Performance Tested MethodSM certification for simultaneous determination and confirmation of aflatoxins (AF) B1, B2, G1, G2, and M1; deoxynivalenol (DON), fumonisins B1, B2, and B3; ochratoxin A (OTA); T-2; HT-2; and zearalenone (ZON) from corn, wheat, cereal-based baby food (with and without dairy ingredients), paprika, chili powder, and animal feed.

METHODS

A single extraction method using acetonitrile-water (1 + 1, by volume) was used for all matrixes. The method developer validated all matrixes and an independent laboratory verified method performance on corn and animal feed. Data were analyzed for recovery, repeatability precision, LOD, LOQ, confirmation of identity, and method selectivity.

RESULTS

Recovery (72-138%) and repeatability (0.46-24%), with the exception of sporadic data points, were within acceptance criteria. LOQ was estimated as AFB1 0.018-0.32 μg/kg, AFB2 0.037-0.28 μg/kg, AFG1 0.019-0.14 μg/kg, AFG2 0.036-0.28 μg/kg, DON 4.0-75 μg/kg, fumonisin B1 4.9-37 μg/kg, fumonisin B2 4.0-32 μg/kg, fumonisin B3 2.0-16 μg/kg, OTA 0.15-4.4 μg/kg, T-2 0.5-7.5 μg/kg, HT-2 0.70-7.5 μg/kg, and ZON 1.3-7.2 μg/kg, depending on matrix. Method performance was verified with reference and QC materials. Selectivity and confirmation of identity were also demonstrated.

CONCLUSION

The 11+Myco MS-PREP IAC with LC-MS/MS method demonstrated acceptable performance for simultaneous determination of 12 mycotoxins in seven matrixes.

HIGHLIGHT

The data were reviewed by the AOAC Performance Tested MethodsSM Program and approval was granted for certification of the 11+Myco MS-PREP Method as PTM 112401.

摘要

背景

11 + Myco MS - PREP®免疫亲和柱（IAC）含有针对目标毒素的单克隆抗体凝胶悬浮液。样品提取后，IAC用于在通过液相色谱 - 串联质谱法（LC - MS/MS）分析之前对霉菌毒素进行净化和浓缩。

目的

本研究评估了采用IAC与LC - MS/MS方法进行性能测试方法SM认证，以同时测定和确证玉米、小麦、谷物类婴儿食品（含或不含乳制品成分）、辣椒粉、辣椒粉和动物饲料中的黄曲霉毒素（AF）B1、B2、G1、G2和M1；脱氧雪腐镰刀菌烯醇（DON）、伏马菌素B1、B2和B3；赭曲霉毒素A（OTA）；T - 2；HT - 2；以及玉米赤霉烯酮（ZON）。

方法

对所有基质采用乙腈 - 水（体积比1 + 1）的单一提取方法。方法开发者对所有基质进行了验证，一个独立实验室对玉米和动物饲料的方法性能进行了验证。分析数据以确定回收率、重复性精密度、检测限、定量限、身份确证和方法选择性。

结果

除个别数据点外，回收率（72 - 138%）和重复性（0.46 - 24%）在可接受标准范围内。定量限估计为AFB1 0.018 - 0.32 μg/kg、AFB2 0.037 - 0.28 μg/kg、AFG1 0.019 - 0.14 μg/kg、AFG2 0.036 - 0.28 μg/kg、DON 4.0 - 75 μg/kg、伏马菌素B1 4.9 - 37 μg/kg、伏马菌素B2 4.0 - 32 μg/kg、伏马菌素B3 2.0 - 16 μg/kg、OTA 0.15 - 4.4 μg/kg、T - 2 0.5 - 7.5 μg/kg、HT - 2 0.70 - 7.5 μg/kg和ZON 1.3 - 7.2 μg/kg，具体取决于基质。用参考物质和质量控制物质验证了方法性能。还证明了方法的选择性和身份确证。