Institute of Agri-products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China.
Institute of Agri-products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Poult Sci. 2020 Jun;99(6):3280-3285. doi: 10.1016/j.psj.2020.01.031. Epub 2020 Mar 18.
Defeathering with rosin results in rosin residue in duck skin, which may present as potential risk to human health. Dehydroabietic acid (DHAA) is a major component of rosin. An indirect competitive enzyme-linked immunosorbent assay (ELISA) was developed for determination of DHAA in duck skin. A set of parameters was optimized, including coating antigen concentration, dilution of antiserum, dilution of HRP-IgG antibody, incubation time, and temperature for antigen reaction with antiserum. The indirect competitive ELISA yielded an excellent specificity against DHAA with low cross-reactivity toward other resin acids. The limit of detection and the working concentration range of DHAA in duck skin were 16.4 ng/g and from 40 to 8,060 ng/g, respectively. The indirect competitive ELISA was applied to the determination of DHAA in duck skin samples spiked with DHAA at different contents, and recoveries were found between 78.2 and 97.2%. Finally, DHAA contents in 32 duck samples were quantified by the indirect competitive ELISA and high performance liquid chromatography-fluorescence detector (HPLC-FLD) method. No significant difference was found between DHAA concentrations from indirect competitive ELISA and HPLC-FLD method for all samples, which indicated the indirect competitive ELISA established in this article was of the same accuracy as the HPLC-FLD method. The indirect competitive ELISA was simple, rapid, and reliable, which could be used to identify the duck carcasses defeathered with rosin in the market.
松香褪毛会导致鸭皮上残留松香，这可能对人类健康构成潜在风险。脱氢枞酸（DHAA）是松香的主要成分之一。本研究建立了一种间接竞争酶联免疫吸附法（ELISA）用于检测鸭皮中的 DHAA。对包括包被抗原浓度、抗血清稀释度、HRP-IgG 抗体稀释度、抗原与抗血清孵育时间和温度等参数进行了优化。间接竞争 ELISA 对 DHAA 具有良好的特异性，对其他树脂酸的交叉反应性较低。DHAA 在鸭皮中的检测限和工作浓度范围分别为 16.4ng/g 和 40-8060ng/g。该间接竞争 ELISA 已成功应用于不同含量 DHAA 加标鸭皮样品的测定，回收率在 78.2-97.2%之间。最后，采用间接竞争 ELISA 和高效液相色谱-荧光检测器（HPLC-FLD）法对 32 个鸭皮样品中的 DHAA 含量进行了定量分析。两种方法检测的所有样品中 DHAA 浓度均无显著差异，表明本文建立的间接竞争 ELISA 与 HPLC-FLD 法具有相同的准确性。该间接竞争 ELISA 简单、快速、可靠，可用于鉴定市场上松香褪毛的鸭胴体。
