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一种通过酶解和分子印迹固相萃取(MISPE)快速检测猪肌肉、肾脏和肝脏中赭曲霉毒素A的高效液相色谱-荧光检测法。

A rapid HPLC-FLD method for Ochratoxin A detection in pig muscle, kidney, liver by using enzymatic digestion with MISPE extraction.

作者信息

Luci Giacomo

机构信息

Department of Clinical and Experimental Medicine, University of Pisa, Via Roma 55, 56126 Pisa, Italy.

出版信息

MethodsX. 2020 Mar 19;7:100873. doi: 10.1016/j.mex.2020.100873. eCollection 2020.

Abstract

Ochratoxin A (OTA) is a mycotoxin produced as a secondary metabolite by various Aspergillus and Penicillium species with nephrotoxic, carcinogenic, immunotoxic and teratogenic effects. OTA has been found in several food commodities, including cereals, beer, wine and spices. OTA can also be present in animal products (especially in pig derived products) as a result of carryover from contaminated feed. Permitted level of 1 µg/kg OTA in pig meat or pig-derived products was set in Italy, as in other countries. Key parameters which affected MISPE, should be described such as extraction efficiency and were optimized, analyzed by an isocratic HPLC-FLD method. Under the optimized conditions, for all analyzed matrices mean recovery was > 89%. Method can be applied as alternative routine procedure to detect OTA presence in pig products. Points:  *Aim of the study was to develop and validate a quantitative HPLC-FLD method based on MISPE with complex solid matrices (edible tissues) followed by chromatographic analysis.  *The new method was developed and validated in pig complex matrix and is very sensitive LOD and LOQ respectively 0.001 and 0.003 µg/kg.  *This method is relatively simple to use and with good performances. Was possible to reuse MISPE column with a "regeneration" solution, until to 7 times.

摘要

赭曲霉毒素A(OTA)是由多种曲霉属和青霉属真菌产生的一种次生代谢产物霉菌毒素,具有肾毒性、致癌性、免疫毒性和致畸性。OTA已在多种食品中被发现,包括谷物、啤酒、葡萄酒和香料。由于受污染饲料的残留,OTA也可能存在于动物产品中(尤其是猪肉衍生产品)。与其他国家一样,意大利规定猪肉或猪肉衍生产品中OTA的允许含量为1微克/千克。应描述影响分子印迹固相萃取(MISPE)的关键参数,如萃取效率,并通过等度高效液相色谱-荧光检测法(HPLC-FLD)进行优化和分析。在优化条件下,所有分析基质的平均回收率均>89%。该方法可作为检测猪产品中OTA存在情况的替代常规方法。要点:*本研究的目的是开发并验证一种基于MISPE的定量HPLC-FLD方法,该方法用于复杂固体基质(可食用组织),随后进行色谱分析。*新方法在猪的复杂基质中开发并验证,灵敏度非常高,检测限(LOD)和定量限(LOQ)分别为0.001和0.003微克/千克。*该方法使用相对简单,性能良好。可以用“再生”溶液重复使用MISPE柱,最多可重复使用7次。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6e17/7153302/b482597068b5/fx1.jpg

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