Yao Kai, Wen Kai, Shan Wenchong, Xie Sanlei, Peng Tao, Wang Jianyi, Jiang Haiyang, Shao Bing
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing 100193, People's Republic of China; Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control, 100013, People's Republic of China.
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing 100193, People's Republic of China; Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing 100193, People's Republic of China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Mar 30;1080:50-58. doi: 10.1016/j.jchromb.2018.02.013. Epub 2018 Feb 13.
An immunoaffinity clean-up material based on a monoclonal antibody (mAb) has been prepared for concentrating and purifying bisphenol A (BPA) in 14 kinds of foodstuffs at trace level. Haptens and immunogen of bisphenol A have been synthesized and comprehensively characterized. An mAb towards BPA was prepared and cross-reactivities with 14 BPA analogues were below 5%. The prepared antibody was coupled to N-hydroxysuccinimide-activated Sepharose 4B to manufacture an immunoaffinity column (IAC), which was applied to purify BPA in 14 kinds of foodstuffs. The analyte was then detected by means of ultra-high-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Under the optimized conditions, compared with two traditional SPE clean-up methods, the IAC showed better selectivity (matrix effect <16.8%) and higher sensitivity. The limits of detection for BPA in 14 kinds of foodstuffs ranged from 0.001 μg L to 0.01 μg kg, and the limits of quantification were in the range from 0.003 μg L to 0.04 μg kg. The recoveries of BPA from spiked samples ranged from 82.0% to 104.9%, with RSDs below 13.8%. Besides, the IAC exhibited good reusability, with 40% column capacity remaining and no significant loss of recovery after 25 application cycles in real sample detection. These results demonstrated that the developed IAC-UPLC-MS/MS approach has wide applicability for purifying and detecting BPA in various foodstuffs.
已制备了一种基于单克隆抗体（mAb）的免疫亲和净化材料，用于浓缩和纯化14种食品中痕量水平的双酚A（BPA）。合成并全面表征了双酚A的半抗原和免疫原。制备了针对双酚A的单克隆抗体，其与14种双酚A类似物的交叉反应率低于5%。将制备的抗体与N-羟基琥珀酰亚胺活化的琼脂糖4B偶联，制成免疫亲和柱（IAC），用于纯化14种食品中的双酚A。然后通过超高效液相色谱串联质谱（UPLC-MS/MS）对分析物进行检测。在优化条件下，与两种传统的固相萃取净化方法相比，免疫亲和柱表现出更好的选择性（基质效应<16.8%）和更高的灵敏度。14种食品中双酚A的检测限为0.001μg/L至0.01μg/kg，定量限为0.003μg/L至0.04μg/kg。加标样品中双酚A的回收率为82.0%至104.9%，相对标准偏差低于13.8%。此外，免疫亲和柱具有良好的可重复使用性，在实际样品检测中经过25个应用循环后，柱容量仍保留40%，回收率无显著损失。这些结果表明，所开发的免疫亲和柱-超高效液相色谱串联质谱方法在纯化和检测各种食品中的双酚A方面具有广泛的适用性。
