在海洋藥用資源的探勘過程中，源自軟珊瑚之活性二次代謝產物的含量與特徵往往與其生長環境的物理與化學因子變化密切相關。因此嘗試探討這些由於生長環境所造成的代謝體變化，將能助益海洋藥物之發展。本實驗利用高效液相層析指紋圖譜 (high-performance liquid chromatography) 分析野外型及養殖型柔軟指型軟珊瑚 Sinularia flexibilis 之化學成分組成，探討其在不同環境、季節的二次代謝物差異。首先我們篩選出四個由柔軟指型軟珊瑚的活性成分作為比較依據，根據實驗結果指出四個活性成分的滯留時間(retention time)皆座落於24-25min這個區間，因此我們將其作為判斷不同來源樣品是否具有活性指標成分之依據。由採集自不同年份之野外型珊瑚樣品的指紋圖譜結果可以發現每一年的樣品皆具有活性成分，樣品間並無明顯差距，而採集自不同季節在相同養殖環境下之柔軟指型樣品的指紋圖譜結果發現活性成分較少，並且在不同季節所採集之樣品存在明顯差異。接著我們將同一年採集之野外型及養殖型珊瑚樣品之指紋圖譜進行比對可以發現珊瑚在人工養殖環境下，活性化合物有明顯的遞減現象，因此我們推斷其可能原因為在人工養殖環境中，珊瑚群體缺乏競爭及掠食者干擾而導致活性化合物明顯減少，而野外珊瑚樣品則因外在環境因素複雜而穩定產生活性化合物。總結以上，進一步找尋影響珊瑚活性二次代謝產物含量的變因，為發展養殖型軟珊瑚作為藥用成分量產之重要課題。

During the exploration of marine medicinal resources, the contents and chemical patterns of bioactive secondary metabolites derived from soft corals are often closely related to changes of physical and chemical factors in their growth environment. Therefore, to explore these metabolomic changes will promote the marine drugs development. In the current study, high-performance liquid chromatography (HPLC) fingerprinting was applied to analyze the chemical compositions of wild-type and aquacultured soft coral Sinularia flexibilis, and to explore their metabolomic differences in various environments and seasons. First, we picked up four bioactive compounds from S. flexibilis, which retained in the range of 24-25 min in HPLC fingerprint, as a basis for following comparison. The results indicated that the contents of four bioactive compounds were identical in wild-type S. flexibilis (collected in different years), but rare in aquacultured one. Moreover, the seasonal variances in aquacultured S. flexibilis were found to be significant and decrease year by year. Therefore, we inferred that the lack of competition was appeared in artificial breeding environment, while wild-type S. flexibilis stably produced bioactive compounds due to complex external environmental factors. In summary, further research on the variable factors of secondary metabolites from soft corals is an important issue for the future development of pharmaceutical mass production.

第一章、前言 1
第一節、野外型柔軟指型軟珊瑚(Sinularia flexibilis)所含煙草烷類(cembrane type)天然物的文獻回顧 2
第二節、養殖型柔軟指型軟珊瑚(Sinularia flexibilis)所含煙草烷類(cembrane type)天然物的文獻回顧 17
第三節、養殖型柔軟指型軟珊瑚(Sinularia flexibilis)所申請的專利 20
第四節、研究動機 21
第二章、生物材料及研究方法 23
第一節、生物樣品採集 23
第二節、珊瑚養殖方法 24
第三節、樣品分類地位 25
第四節、生物樣品的萃取、分離及化合物的純化 26
第五節、4個目標化合物之生物活性回顧 27
第六節、以高效液相層析儀 (high-performance liquid chromatography) 進行建立珊瑚化學指紋圖譜 29
第三章、實驗設備儀器及材料、相關溶劑 31
第一節、實驗設備儀器 31
第二節、層析材料 31
第三節、相關溶劑 32
第四章、結果與討論 33
第一節、標準品指紋圖譜 33
第二節、野外型柔軟指型軟珊瑚指紋圖譜 35
第三節、養殖型柔軟指型軟珊瑚指紋圖譜 37
第四節、野外型與養殖型柔軟指型軟珊瑚指紋圖譜比較 39
第五章、討論與未來展望 41
參考文獻 43
附錄一 49
附錄二 50
附錄三 51
附錄四 52
附錄五 (珊瑚指紋圖譜分析) 53

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