A multiplex quadruple-label time-resolved fluorescence immunochromatographic assay (QL-TRFICA) for simultaneous quantitative detection of aflatoxin B₁ (AFB₁), zearalenone (ZEN) and ochratoxin A (OTA) in grains was established in this paper. This assay was developed based on quadruple-label immunoprobes, one of which was a chicken IgY-specific antibody coupled with time-resolved fluorescent nanobeads (TRFN) for the test C line signal while the other three were specific to AFB₁, ZEN and OTA monoclonal antibodies coupled with TRFN for the test T line signal. Quantitative relationships between the concentrations of the three mycotoxins and T/C ratios were established to determine the analyte concentration through a portable multi-channel fluorescence reader. The limits of detection (LODs) for AFB₁, ZEN and OTA were 0.04 μg kg⁻¹, 0.20 μg kg⁻¹ and 0.10 μg kg⁻¹ in 6 grains (corn, soybean, sorghum, wheat, rice and oats), respectively. Recoveries ranged from 71.60% to 119.98% at fortified concentrations of LOD, 2LOD and 4LOD, with a coefficient of variation <15%. Analysis of field grain samples by QL-TRFICA was in accordance with that of LC-MS/MS. Therefore, this proposed multiplex QL-TRFICA was a promising method for on-site screening of multiple mycotoxins in grains.

中文翻译：

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四标记时间分辨荧光免疫色谱分析法同时定量测定谷物中的三种霉菌毒素

建立了同时定量检测谷物中黄曲霉毒素B ₁（AFB ₁），玉米赤霉烯酮（ZEN）和曲霉毒素A（OTA）的多重四重标记时间分辨荧光免疫层析法（QL-TRFICA）。此测定法是基于四重免疫探针开发的，其中一种是鸡IgY特异性抗体，结合时间分辨荧光纳米珠（TRFN）用于测试C线信号，而其他三种是AFB ₁特异性的，ZEN和OTA单克隆抗体与TRFN偶联用于测试T线信号。建立了三种霉菌毒素浓度与T / C比之间的定量关系，以通过便携式多通道荧光读取器确定分析物浓度。检测的对AFB限（检测限）₁，ZEN和OTA是0.04微克千克^-1，0.20微克千克^-1和0.10微克千克^-1分为6种谷物（玉米，大豆，高粱，小麦，大米和燕麦）。强化浓度的LOD，2LOD和4LOD的回收率范围为71.60％至119.98％，变异系数<15％。QL-TRFICA对田间谷物样品的分析与LC-MS / MS一致。因此，该拟议的多重QL-TRFICA是现场筛选谷物中多种霉菌毒素的有前途的方法。