Development of a sensitive and reliable high performance liquid chromatography method with fluorescence detection for high-throughput analysis of multi-class mycotoxins in Coix seed
Development of a sensitive and reliable high performance liquid chromatography method with fluorescence detection for high-throughput analysis of multi-class mycotoxins in Coix seed
复制标题
开发灵敏可靠的荧光检测高效液相色谱方法，用于薏苡仁中多类霉菌毒素的高通量分析
DOI：
10.1016/j.aca.2013.08.042
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发表时间：
2013-10-17
影响因子：
6.2
通讯作者：
Yang, Mei-Hua
中科院分区：
文献类型：
--
作者：
Kong, Wei-Jun;Li, Jun-Yuan;Yang, Mei-Hua
薏苡仁是一种可食用和药用的植物，易被一组以上的真菌毒素污染，对人体健康有潜在的危害。建立了一种可靠、灵敏的方法，可同时测定国内销售的10批薏苡仁中7种真菌毒素(黄曲霉毒素B-1、B-2、G（1)、G(2)、玉米赤霉烯酮、α -玉米赤霉烯醇、β -玉米赤霉烯醇）。该方法采用甲醇/水（80/20）快速超声辅助固液萃取（USLE），免疫亲和柱（IAC）净化，在线光化学衍生化（PCD），高效液相色谱-荧光检测（HPLC-FLD）。对提取、清理、分离和检测条件进行了优化，提高了样品通量，实现了快速分离和灵敏检测。通过分析以三种不同浓度加标的样品对7种真菌毒素进行方法验证。加样回收率为73.5% ~ 107.3%，相对标准偏差(rsd) < 7.7%。日内和日间精密度（以rsd表示）对所有研究分析物均低于4%。检测限和定量限分别为0.01 ~ 50.2 μ g kg（-1）和0.04 ~ 125.5 μ g kg(-1)，均低于欧盟规定的真菌毒素耐受水平。检测呈阳性的样品进一步用高效液相色谱串联电喷雾质谱法进行验证分析。这是首次应用USLE-IAC-HPLC-PCD-FLD检测薏苡仁中多类真菌毒素的情况。(c) 2013 Elsevier B.V.版权所有
As an edible and medicinal plant, Coix seed is readily contaminated by more than one group of mycotoxins resulting in potential risk to human health. A reliable and sensitive method has been developed to determine seven mycotoxins (aflatoxins B-1, B-2, G(1), G(2), zearalenone, alpha-zearalenol, and beta-zearalenol) simultaneously in 10 batches of Coix seed marketed in China. The method is based on a rapid ultrasound-assisted solid-liquid extraction (USLE) using methanol/water (80/20) followed by immunoaffinity column (IAC) clean-up, on-line photochemical derivatization (PCD), and high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD). Careful optimization of extraction, clean-up, separation and detection conditions was accomplished to increase sample throughput and to attain rapid separation and sensitive detection. Method validation was performed by analyzing samples spiked at three different concentrations for the seven mycotoxins. Recoveries were from 73.5% to 107.3%, with relative standard deviations (RSDs) lower than 7.7%. The intra-and inter-day precisions, expressed as RSDs, were lower than 4% for all studied analytes. Limits of detection and quantification ranged from 0.01 to 50.2 mu g kg(-1), and from 0.04 to 125.5 mu g kg(-1), respectively, which were below the tolerance levels for mycotoxins set by the European Union. Samples that tested positive were further analyzed by HPLC tandem electrospray ionization mass spectrometry for confirmatory purposes. This is the first application of USLE-IAC-HPLC-PCD-FLD for detecting the occurrence of multi-class mycotoxins in Coix seed. (c) 2013 Elsevier B.V. All rights reserved.