乌头属药材中单酯型乌头碱的ELISA法测定

Determination of Mono-ester Alkaloids in Aconitum Plants by ELISA

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0．005～5gg／ml，检测限为139pg，定量限为575Pg，平均回收率97．31％（RSD≤1．68％）。ELISA法的灵敏度较HPLC法高约112倍。

The hapten, mono benzoylmesaconine-3-yl glutarate （BM-GA）, was synthesized by introduction of a glutaryl group to benzoylmesaconine （BM）. The derivative was then conjugated with bovine serum albumin （BSA） by the active ester method to form the artificial antigen （BM-GA-BSA）. A polyclonal antibody （PAb） was produced from immunized New Zealand rabbits. It had been proved that the PAb had high specificity with mono-ester alkaloids. An enzyme-linked immunosorbent assay （ELISA） method was established for the determination of mono-ester alkaloids in Aconitum plants. The linear range of mono-ester alkaloids was 0.005-5 μg/ml. The limit of detection （LOD） was 139 pg, the limit of quantity （LOQ） was 575 pg, and the average recovery rate was 97.31%, with RSDs no more than 1.68 %. The sensitivity of ELISA was approximately 112 times higher than that of HPLC.