Center of Excellence in Food Safety Research, Faculty of Food Science and Technology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2010 Dec;27(12):1683-93. doi: 10.1080/19440049.2010.514951.
Method validation for quantitative analysis of aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZEA) in cereals using HPLC with fluorescence detector (FLD) is described. Mycotoxins were extracted with methanol : water (80 : 20) and purified with a multifunctional AOZ immunoaffinity column before HPLC analysis. The validation of the analytical method was performed to establish the following parameters: specificity, selectivity, linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision (within- and between-day variability), stability, robustness, measurement of performance, and measurement of uncertainty. Calibration curves were linear (r > 0.999) over the concentration range, from the LOQ to 26, 40 and 400 ng/g for AFs, OTA and ZEA, respectively. LOD and LOQ were 0.0125 and 0.05 ng/g for aflatoxin B1 (AFB1) and G1 (AFG1), 0.0037 and 0.015 ng/g for aflatoxin B2 (AFB2) and G2 (AFG2), as well as 0.05 and 0.2 ng/g for OTA and 0.5 and 2 ng/g for ZEA, respectively. The mean recovery values were 77-104% for different concentrations of AFs, OTA and ZEA in spiked cereal samples. Both intra- and inter-day accuracy and precision were within acceptable limits. This method was successfully applied for the simultaneous determination of mycotoxins for 60 cereal samples collected from Malaysian markets. Fifty per cent of the cereal samples were contaminated with at least one of these mycotoxins, at a level greater than the LOD. Only one wheat sample and two rice samples were contaminated with levels greater than the European Union regulatory limits for AFs and OTA (4 and 5 ng/g). The means and ranges of mycotoxins obtained for the cereal samples were 0.4 ng/g and 0.01-5.9 ng/g for total AFs; 0.18 ng/g and 0.03-5.3 ng/g for OTA; and 2.8 ng/g and 2.4-73.1 ng/g for ZEA, respectively. The results indicate that the method is suitable for the simultaneous determination of AFs, OTA and ZEA in cereals and is suitable for routine analysis.
本文描述了使用高效液相色谱法（HPLC）结合荧光检测器（FLD）对谷物中的黄曲霉毒素（AFs）、赭曲霉毒素 A（OTA）和玉米赤霉烯酮（ZEA）进行定量分析的方法验证。真菌毒素用甲醇：水（80：20）提取，并用多功能 AOZ 免疫亲和柱纯化，然后进行 HPLC 分析。对分析方法进行验证，以确定以下参数：特异性、选择性、线性、检测限（LOD）和定量限（LOQ）、准确度、精密度（日内和日间变异性）、稳定性、稳健性、性能测量和不确定度测量。AFs、OTA 和 ZEA 的校准曲线在浓度范围内从 LOQ 到 26、40 和 400ng/g 呈线性（r＞0.999）。AFB1 和 AFG1 的 LOD 和 LOQ 分别为 0.0125 和 0.05ng/g，AFB2 和 AFG2 分别为 0.0037 和 0.015ng/g，OTA 和 ZEA 分别为 0.05 和 0.2ng/g。在添加不同浓度的 AFs、OTA 和 ZEA 的谷物样品中，平均回收率为 77-104%。不同浓度的 AFs、OTA 和 ZEA 的日内和日间准确度和精密度均在可接受范围内。该方法成功应用于马来西亚市场采集的 60 个谷物样品中真菌毒素的同时测定。50%的谷物样品受到至少一种真菌毒素的污染，污染水平高于 LOD。只有一个小麦样品和两个大米样品受到 AFs 和 OTA（4 和 5ng/g）欧盟法规限量以上水平的污染。从谷物样品中获得的真菌毒素的平均值和范围分别为：总 AFs 为 0.4ng/g，0.01-5.9ng/g；OTA 为 0.18ng/g，0.03-5.3ng/g；ZEA 为 2.8ng/g，2.4-73.1ng/g。结果表明，该方法适用于谷物中 AFs、OTA 和 ZEA 的同时测定，适合常规分析。
