| Alkaloids that are the main active components in Sophora alopecuroides. L havevaluable applications in agriculture and medicine, therefore more attention has paid on theirdetection method. It's necessary to eastabish a rapid, sensitive and efficient method fordetecting multiple alkaloids. This dissertation focused on application for indirect competitiveenzyme-linked immunosorbent assay (IC-ELISA). The main results are as follows:1. The hapten13-aminomatrine(ST) was designed and synthesized successfully. ST wassynthesized by using sophocarpine through nucleophilic addition, diazotization andhydrogenation then characterized by MS and NMR. The Glutaraldehyde method was used toconjugate hapten to BSA and OVA, the artifical antigen were verified by SDS-PAGE, UV andIR. The ratio came to9:1and4:1, respectively, which were confimed by2,4,6-trinitrebenzeneSulfonicacid method.2. One type of anti-matrine polyclonal antibodies were obtained and the highest titer ofanti-matrine serum was128000.3. An IC-ELISA for matrine was established. its linear equation was y=13.431x+37.718(r=0.9887)in the range of0~100μg/mL. The limit of detection(LOD) IC10and IC50formatrine was0.086μg/ml and8.212μg/mL. The average recovery of matrine in tap waterdetermined by IC-ELISA is76.93~96.59%in the added range of0~50ug/mL. This assayshowed high detected ability with the structrally closely related compounds, includingsophocarpine, sophoridine and oxymatrine.4A detection method for matrine by high performance liquid chromatography (HPLC)was established. There was a good linear relationship between peak area and concentration ofanalytes over the concentration range of0.25~5μg/mL(r=0.9975). A good correlationbetween matrine concentration measured by HPLC and IC-ELISA was obtained (r=0.9982)from linear regression analysis.5HPLC and IC-ELISA method were used to analysed the content of matrine in0.5%matrine aqueous solution. Those results indicate that the ELISA could be a supplementanalytical tool for monitor matrine. |
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