Center of Excellence for 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. 2011 Apr;28(4):494-501. doi: 10.1080/19440049.2010.551547. Epub 2011 Feb 17.
A new method for the simultaneous quantification of 12 mycotoxins was developed and optimized using reverse phase high performance liquid chromatography (RP-HPLC) with a photodiode array (PDA) and fluorescence detector (FLD), a photochemical reactor for enhanced detection (PHRED) and post-column derivatization. The mycotoxins included aflatoxins (AFB(1), AFB(2), AFG(1), and AFG(2)), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisins (FB(1), FB(2), and FB(3)), T-2 and HT-2 toxins. A double sample extraction with a phosphate-buffered saline solution (PBS) and methanol was used for co-extraction of mycotoxins, and a multifunctional immunoaffinity column was used for cleanup. Optimum conditions for separation of the mycotoxins were obtained to separate 12 mycotoxins in FLD and PDA chromatograms with a high resolution. The method gave recoveries in the range 72-111% when applied to spiked corn samples. The limits of detection (LOD) were 0.025 ng/g for AFB(1) and AFG(1), 0.012 ng/g for AFB(2) and AFG(2), 0.2 ng/g for OTA, 1.5 ng/g for ZEA, 6.2 ng/g for FB(1), FB(3) and HT-2 toxin, 9.4 ng/g for FB(2) and T-2 toxin, and 18.7 ng/g for DON. In addition, the limits of quantification (LOQ) ranged from 0.04 ng/g for AFB(2) and AFG(2) to 62 ng/g for DON. The method was successfully applied to the determination of these mycotoxins in 45 cereal samples obtained from the Malaysian market. The results indicated that the method can be applied for the multi-mycotoxin determination of cereals.
建立并优化了一种新的同时定量检测 12 种真菌毒素的方法，该方法采用反相高效液相色谱（RP-HPLC）结合光电二极管阵列（PDA）和荧光检测器（FLD）、光化学增强检测（PHRED）和柱后衍生化。所检测的真菌毒素包括黄曲霉毒素（AFB(1)、AFB(2)、AFG(1)和 AFG(2)）、赭曲霉毒素 A（OTA）、玉米赤霉烯酮（ZEA）、脱氧雪腐镰刀菌烯醇（DON）、伏马菌素（FB(1)、FB(2)和 FB(3)）、T-2 和 HT-2 毒素。采用磷酸盐缓冲盐水（PBS）和甲醇的双样本提取法来同时提取真菌毒素，并用多功能免疫亲和柱进行净化。通过优化分离条件，在 FLD 和 PDA 色谱图中可以实现 12 种真菌毒素的高分辨率分离。当应用于玉米加标样品时，该方法的回收率在 72%-111%之间。AFB(1)和 AFG(1)的检测限（LOD）为 0.025ng/g，AFB(2)和 AFG(2)的 LOD 为 0.012ng/g，OTA 的 LOD 为 0.2ng/g，ZEA 的 LOD 为 1.5ng/g，FB(1)、FB(3)和 HT-2 毒素的 LOD 为 6.2ng/g，FB(2)和 T-2 毒素的 LOD 为 9.4ng/g，DON 的 LOD 为 18.7ng/g。此外，定量限（LOQ）范围从 AFB(2)和 AFG(2)的 0.04ng/g 到 DON 的 62ng/g。该方法成功地应用于从马来西亚市场获得的 45 种谷物样品中这些真菌毒素的测定。结果表明，该方法可用于谷物中多种真菌毒素的检测。
