本研究的目的在於建立一套適用於紅麴中內生毒素橘黴素(Citrinin)之酵素免疫化學分析法以求有效定量紅麴食品Citrinin的含量，並且初步探討Citrinin的細胞毒性。實驗以紐西蘭大白兔來生產對於Citrinin具有專一性的多株抗體，首先將Citrinin以化學接合方法分別接到牛血清蛋白(Bovine serum albumin, BSA)與血藍蛋白(Keyhole limpet hemocyanin, KLH)上使其成為良好的免疫原，然後將這些接合物免疫到兔子體內並收集血清以進行抗體效價測試，由非直接酵素免疫分析法的結果顯示，兩種不同接合物所得到的抗體效價均隨著週次增加而升高，但是競爭型酵素免疫分析法的分析結果則顯示外加毒素並無法有效取代毒素酵素標記鍵結到抗體上。另一方面，利用人類(293)及犬類腎臟細胞株(MDCK)來探討Citrinin對標的細胞株的生化毒性時，發現經由不同濃度之Citrinin處理72小時後，與對照組相比之50%致死濃度約為80μM，人類及犬類腎臟細胞具有相似的敏感度。實驗結果顯示Citrinin的細胞毒性應非經由細胞凋亡(Apoptosis)的途徑。
The objectives of this research were to establish an enzyme-linked immunosorbent assay for determination of citrinin in Monascus. Spp. and cytotoxicity test of citrinin. Polyclonal antibodies for citrinin were generated from rabbits after immunizing the animals with citrinin conjugated with BSA and KLH, respectively. A competitive indirect enzyme-linked immunosorbent assay (ciELISA) was used for the characterization of the antibodies. The antibody titers increased progressively following the week. A competitive direct enzyme-linked immunosorbent assay (cd ELISA) was established for analysis of the toxin. However, the marker antigen (CTN-HRP conjugates) could not displace the free toxin in the cd ELISA. The human cell line H293 and dog kidney cell line (MDCK) were used to test the biochemical toxicities. The results showed that the LD/sub 50/ for citrinin on these two cell lines was about 80.mu.M after treatment of 72 hr. The results indicated that the cytotoxicity of citrinin is not through the apoptosis pathway.
