酶联免疫吸附测定法检测白条鸭表皮组织中的脱氢松香酸

Determination of Dehydroabietic Acid in Duck Skin by Enzyme-Linked Immunosorbent Assay

通过优化抗原包被质量浓度、抗体稀释倍数及二抗稀释倍数等参数,建立白条鸭表皮组织中脱氢松香酸(dehydroabietic acid,DHAA)含量的间接竞争酶联免疫吸附测定(enzyme linked immunosorbent assay,ELISA)方法。白条鸭表皮组织中的DHAA用甲醇提取,经磷酸盐缓冲液稀释,使用酶标板进行检测。结果表明:最佳抗原包被质量浓度为1.0μg/mL,最佳抗体稀释倍数为1∶6400,最佳二抗稀释倍数为1∶10000;ELISA方法的检测限及检测范围分别为41.0 ng/g和100.0~20150.0 ng/g;在100~5000 ng/g添加量范围内,DHAA回收率为78.2%~97.2%;实际样品分析结果显示,市场流通领域中白条鸭表皮组织中DHAA的检出率达到87%,含量范围为118.6~1199.2 ng/g。

An indirect competitive enzyme-linked immunosorbent assay(ELISA)was developed for determination of dehydroabietic acid(DHAA)in duck skin.DHAA in duck skin samples was extracted with methanol,followed by dilution with phosphate buffer saline(PBS),and determined by an ELISA reader.Results indicated that the optimal coating antigen concentration and the optimal dilution of antiserum were found to be 1.0μg/mL and 1:6400,respectively,while the optimal dilution of HRP-IgG antibody was 1:10000,the limit of detection and the working concentration range were 41.0 ng/g and from 100.0 to 20150.0 ng/g respectively.The recoveries from spiked samples at concentration levels of 100–5000 ng/g were in the range of 78.2%–97.2%.When the method was applied to real samples,DHAA was detectable in up to 87%of the samples at levels of 118.6–1199.2 ng/g.