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基于 BSA 的样品净化柱用于葡萄酒中赭曲霉毒素 A 的测定:方法开发与验证。

BSA-based sample clean-up columns for ochratoxin A determination in wine: Method development and validation.

机构信息

CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.

CEB - Centre of Biological Engineering, University of Minho, 4710-057 Braga, Portugal.

出版信息

Food Chem. 2019 Dec 1;300:125204. doi: 10.1016/j.foodchem.2019.125204. Epub 2019 Jul 17.

Abstract

Analytical chromatographic techniques for mycotoxins control are well established, but they often depend on costly immunoaffinity sample clean-up. Serum albumins, particularly that from bovine origin (BSA), have stable binding affinity towards some mycotoxins, and can be cheaper alternative receptors for sample clean-up due to their wide availability. Thus, this work used BSA immobilized in agarose beads as a novel solid-phase extraction method for quantification of ochratoxin A (OTA) in wine. Constructed BSA-agarose columns could extract OTA efficiently from red wine after its dilution (4-fold) in 0.1 M Tris pH 8.0. The method was linear (R = 0.9999) in the OTA concentration range studied (0.05 to 3.0 μg L), with recovery rates above 98%. It also showed low detection (0.017 μg L) and quantification (0.051 μg L) limits. The efficacy of the BSA-based method was further validated by direct comparison with commercial immunoaffinity columns. Portuguese wines analyzed by both methods had agreeing results.

摘要

用于霉菌毒素控制的分析色谱技术已经成熟,但它们通常依赖于昂贵的免疫亲和样品净化。血清白蛋白,特别是牛源的(BSA),对一些霉菌毒素具有稳定的结合亲和力,并且由于其广泛的可用性,可能是样品净化的更廉价的替代受体。因此,本工作使用琼脂糖珠固定化的 BSA 作为一种新型的固相萃取方法,用于葡萄酒中赭曲霉毒素 A(OTA)的定量分析。构建的 BSA-琼脂糖柱可以在 0.1 M Tris pH 8.0 中稀释(4 倍)后,从红葡萄酒中有效提取 OTA。该方法在研究的 OTA 浓度范围内(0.05 至 3.0 μg L)呈线性(R=0.9999),回收率高于 98%。它还显示出较低的检测(0.017 μg L)和定量(0.051 μg L)限。通过与商业免疫亲和柱的直接比较进一步验证了基于 BSA 的方法的功效。两种方法分析的葡萄牙葡萄酒结果一致。

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