Enzyme-Linked Immunosorbent Assay For The Determination Of Sildenafil In Functional Food

In this paper,the direct competitive enzyme-linked immunosorbent assay (cdELISA) method for the determination of Sildenafil(SIL) in functional food sample had been developed.We synthesized a hapten of SIL and investigated the method of coupling the hapten to carrier albumens.The anti-SIL antibody with high affinity and specificity were obaitned.The cdELISA used in this study exhibited high sensitivity. The IC₅₀ and IC₁₅ were 6±0.01μg/kg and 0.6±0.002μg/kg separately.A simple, rapid and efficient extraction method for the rapid detection of Sildenafil in functional food sample was developed with recoveries between 80～110%.Matrix effects can be avoided when sample diluted with shelter buffer and PBS.In order to validate cdELISA,samples were analyzed by high-performance liquid chromatography(HPLC).Sample extracts were analyzed by ELISA and HPLC, and results obtained by two methods correlated well,the correlation was greater than 0.98.The ELISA method and the rapid extraction method described in this paper is feasible for preparing ELISA kits,which can determine Sildenafil in functional food rapidly.This research will be useful for the development of anti-SIL ELISA kit.