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Determination of Dehydroabietic Acid in Duck Skin by Enzyme-Linked Immunosorbent Assay |
QIU Xinyuan, YAO Zhong, GENG Zhiming, MA Jingjing, LI Pengpeng |
1.College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China; 2.Institute of Agri-Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China |
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Abstract 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 1.0 μg/mL and 1:6 400, respectively, while the optimal dilution of HRP-IgG antibody was 1:10 000, the limit of detection and the working concentration range were 41.0 ng/g and from 100.0 to 20 150.0 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.
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