Cirad, UMR Qualisud, TA B-95/16, 73, rue JF Breton, 34398 Montpellier Cedex 5, France; UMR 95 QualiSud, Université de Montpellier, CIRAD, Montpellier SupAgro, Université d'Avignon, Université de la Réunion, Montpellier, France; Faculty of Biotechnology, Misr University for Science and Technology, 6th 13 October City, Egypt; IES, UMR CNRS 5214, Montpellier University, France.
Cirad, UMR Qualisud, TA B-95/16, 73, rue JF Breton, 34398 Montpellier Cedex 5, France; UMR 95 QualiSud, Université de Montpellier, CIRAD, Montpellier SupAgro, Université d'Avignon, Université de la Réunion, Montpellier, France.
Food Chem. 2020 Apr 25;310:125851. doi: 10.1016/j.foodchem.2019.125851. Epub 2019 Nov 8.
One of the most common mycotoxin contaminating feed and foodstuffs is Ochratoxin A (OTA). OTA has a chronic toxic effect and proved to be mutagenic, nephrotoxic, teratogenic, immunosuppressive, and carcinogenic. Aptamer with their specific affinity for OTA was used in this paper to create an analytical technique. Several methods have been reported for the determination of OTA in foods. However, most of these methods could not be applied to a complex food as green coffee because the interfering native fluorescent products made the quantification very difficult. In this work, we mixed two separations based techniques to identify and quantify OTA in green coffee. Aptamer assisted ultrafiltration as separation technique based on the size of molecules was applied to separate the free OTA, the quantification of OTA was established by a high-performance liquid chromatography (HPLC-FD) with a limit of detection (LOD) of 0.05 ng/mL for OTA. Artificially contaminated green coffee displayed a good range of OTA recoveries up to 97.7%. This method can be applied to the quantitative determination of OTA in green coffee at levels below the maximum levels proposed by the European Commission for green coffee. It also confirm that aptamers can be used as biorecognition element in diagnostic assays with commercial application for mycotoxin analysis.
一种最常见的污染饲料和食品的真菌毒素是赭曲霉毒素 A（OTA）。OTA 具有慢性毒性作用，已被证明具有致突变性、肾毒性、致畸性、免疫抑制性和致癌性。本文利用对 OTA 具有特异性亲和力的适体来创建一种分析技术。已经报道了几种用于测定食品中 OTA 的方法。然而，由于干扰的天然荧光产物使得定量非常困难，大多数这些方法不能应用于复杂的食物如绿咖啡。在这项工作中，我们混合了两种基于分离的技术来识别和定量绿咖啡中的 OTA。基于分子大小的适体辅助超滤作为分离技术，用于分离游离的 OTA，通过高效液相色谱（HPLC-FD）与检测限（LOD）为 0.05ng/mL 的方法来定量 OTA。人工污染的绿咖啡显示出高达 97.7%的 OTA 回收率。该方法可用于定量测定绿咖啡中的 OTA，其水平低于欧盟委员会为绿咖啡规定的最大限量。它还证实适体可以用作诊断测定中的生物识别元件，具有用于真菌毒素分析的商业应用。
