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以苯甲酰新乌头原碱合成人工半抗原3-戊二酰基苯甲酰新乌头原碱,并采用活泼酯法使其与牛血清白蛋白(BSA)结合,制得苯甲酰新乌头原碱免疫抗原。以此抗原免疫新西兰兔,获得苯甲酰新乌头原碱抗体,可特异性识别单酯型乌头碱类生物碱。由此建立了酶联免疫吸附测定法(ELISA)检测乌头属药材中的单酯型乌头碱类生物碱。本法的线性范围为0.005~5?g/ml,检测限为139 pg,定量限为575 pg,平均回收率97.31%(RSD≤1.68%)。ELISA法的灵敏度较HPLC法高约112倍。
The artificial haptens 3-glutaryl benzoyl mesaconitine was synthesized with benzoyl mesaconitine and its active ester method was used to combine with bovine serum albumin (BSA) Aconitine immunogen antigen. New Zealand rabbits were immunized with this antigen to obtain benzoyl-aconitine antibody, which can specifically recognize monoester aconitine alkaloids. Thus, enzyme-linked immunosorbent assay (ELISA) was established for the determination of monoester aconitine alkaloids in the genus Aconitum. The linear range of this method is 0.005 ~ 5 μg / ml, the detection limit is 139 pg, the limit of quantification is 575 pg, the average recovery is 97.31% (RSD ≤ 1.68%). The sensitivity of ELISA method is about 112 times higher than HPLC method.