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Development of indirect competitive ELISA for determination of dehydroabietic acid in duck skin and comparison with the HPLC method

机译：间接竞争力的开发用于测定鸭肌中脱氢酸的脱氢酸与HPLC方法的比较

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摘要

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.
机译：鸭绒松香的耐热物质导致松香残留物，这可能呈现为人类健康的潜在风险。脱氢双皮酸（DHAA）是松香的主要成分。开发间接竞争性酶联免疫吸附试验（ELISA）以测定DuhaA在鸭皮中的DHAA。优化了一组参数，包括涂覆抗原浓度，稀释浸润，稀释HRP-IgG抗体，孵育时间和与抗血清抗原反应的温度。间接竞争力的ELISA对DHAA产生了优异的特异性，对其他树脂酸具有低交叉反应性。鸭皮中DHAA的检测极限和工作浓度范围分别为16.4ng / g，分别为40至8,060ng / g。将间接竞争力的ELISA应用于DhAA在鸭皮样品中的测定，在不同内容物中掺入DHAA，并且在78.2和97.2％之间发现回收率。最后，通过间接竞争力的ELISA和高效液相色谱 - 荧光检测器（HPLC-FLD）方法量化32只鸭样样品中的DHAA含量。从间接竞争力的ELISA和HPLC-FLD方法的DHAA浓度与所有样品的HPLC-FLD方法之间没有显着差异，这表明本文中建立的间接竞争力的ELISA具有与HPLC-FLD方法相同的准确性。间接竞争力的ELISA简单，迅速，可靠，可用于识别在市场上用松香摧毁的鸭子尸体。

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