Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Via Manfredonia 20, 71100 Foggia, Italy.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2009 Oct;26(10):1402-10. doi: 10.1080/02652030903107906.
A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished using a C18 column eluted with an isocratic mobile phase consisting of water, methanol and acetonitrile. The sample preparation required a simple extraction of aflatoxins with MeOH/H2O (80:20, v/v) and a purification step by immunoaffinity column cleanup. The total analysis time, including sample preparation and chromatographic separation, did not exceed 40 min with a run time of 10 min. The on-line photochemical derivatization ensures better results in terms of simplicity, sensitivity and reproducibility with respect to chemical derivatization techniques, and provides an increase of the peak resolution and an extent of automation in comparison with the electrochemical ones. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption was extensively validated following Regulation (EC) No. 882/2004. Detection limits in wheat bran samples of 0.08 μg kg1 for AFB1, 0.02 μg kg1 for AFB2, 0.16 μg kg1 for AFG1 and 0.04 μg kg1 for AFG2 were attained. The method allows high recovery with mean values ranging from 72 to 94% and it satisfies the necessary requirements for sensitivity, linearity, selectivity, precision and ruggedness, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006.
一种用于同时分离和定量测定食品和饲料材料中黄曲霉毒素 B1、B2、G1 和 G2 的灵敏和选择性分析方法已经过验证。该方法基于高效液相色谱法，在线柱后光化学衍生化和荧光检测。黄曲霉毒素的色谱分离使用 C18 柱，以水、甲醇和乙腈组成的等度流动相洗脱。样品制备需要用 MeOH/H2O（80:20，v/v）简单提取黄曲霉毒素，然后通过免疫亲和柱净化进行纯化步骤。包括样品制备和色谱分离在内的总分析时间不超过 40 分钟，运行时间为 10 分钟。在线光化学衍生化确保了与化学衍生化技术相比，在简单性、灵敏度和重现性方面具有更好的结果，并与电化学衍生化相比，提供了峰分辨率的提高和自动化程度的增加。根据法规（EC）No.882/2004 对食品样品和动物饲料用谷物中黄曲霉毒素的测定方法进行了广泛验证。在麦麸样品中，AFB1 的检测限为 0.08 μg kg1，AFB2 为 0.02 μg kg1，AFG1 为 0.16 μg kg1，AFG2 为 0.04 μg kg1。该方法允许高回收率，平均值范围为 72%至 94%，并且满足灵敏度、线性度、选择性、精密度和稳健性的必要要求，证明该方法符合法规（EC）No.401/2006 的规定。
