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建立薯蓣皂甙元的ELISA定量分析方法必须先合成薯蓣皂甙元的全抗原。试验利用薯蓣皂甙元3位上的-OH,在DMAP的催化下,薯蓣皂甙元与丁二酸酐反应,生成薯蓣皂甙元丁二酸单酯,用MSI、R1、H NMR1、3CNMR等方法对产物结构进行了表征。用混合酸酐法制备薯蓣皂甙元与牛血清白蛋白的结合物(DG-HS-BSA),碳二亚胺法制备薯蓣皂甙元与卵清蛋白的结合物(DG-HS-OVA),经TNBS测定,每分子结合物连接的薯蓣皂甙元分子数分别为28.0和8.8。
To establish a ELISA quantitative analysis method for diosgenin, it is necessary to first synthesize the whole antigen of diosgenin. In the experiment, dihydroxysaponin was used in the 3-position of diosgenin, and under the catalysis of DMAP, diosgenin was reacted with succinic anhydride to produce diosgenin monosuccinic acid ester, and the product was analyzed by MSI, R1, H NMR1 and 3CNMR. The structure was characterized. The conjugate of diosgenin and bovine serum albumin (DG-HS-BSA) was prepared by mixed anhydride method, and the conjugate of diosgenin and ovalbumin (DG-HS-OVA) was prepared by carbodiimide method. The number of diosgenin molecules attached per molecule of the conjugate was 28.0 and 8.8, respectively.