臺灣博碩士論文加值系統

蘭胚 (Orchid Embryo)是由蝴蝶蘭屬之蘭花種子於無菌環境下培養30- 45天後形成之擬原球體，由蘭卉生技公司命名為蘭胚。本次研究預實驗中發現蘭胚具有相關抗氧化活性表現，因此進行蘭胚之成分分離及活性實驗。由蘭胚甲醇萃取之氯仿層分離出33個化合物，其中包含6個ionone衍生物、6個單苯環化合物、2個malatic acid衍生物、1個固醇類化合物、1個雙環氧木脂素、4個脂肪酸以及1個三酸甘油酯，另外尚有12個化合物結構待鑑定。其中dimethyl-2-benzyl malate (15)為首次由天然物中分離得到，另外共有19個化合物為首次於蝴蝶蘭屬植物中分離得到。
本次研究中，利用正、逆相兩種不同製程將蘭胚乙醇層萃取物進行成分劃分，並進行抑制α-葡萄糖苷酶活性、清除DPPH自由基能力、抑制共軛雙烯生成、抑制脂質氫過氧化物生成、清除過氧化氫能力、螯合鐵離子能力、抑制酪胺酸酶活性、抑制微脂粒氧化作用以及抑制膠原蛋白酶活性等九種試驗。結果顯示蘭胚乙醇萃取物及其劃分對於清除過氧化氫能力、抑制共軛雙烯生成及抑制微脂粒氧化能力測試中具有良好的功效，另外，在抑制氫過氧化物、螯合鐵離子能力、抑制酪胺酸酶及抑制膠原蛋白酶活性測定中，發現蘭胚萃取物及其各劃分皆具有不錯之活性展現，此外在抑制α-葡萄糖苷酶活性測試中也發現具有極佳之抑制能力。
由上述實驗結果，可提供後續美妝保養產品配方設計、保健食品及其他相關產品之開發與參考。

Orchid Embryo, named by Lan-Hui Biotech Co., is the protocorm-like bodies of Phalaenopsis seeds cultured in sterile MS solid medium for 30~45 days. Based on our preliminary test study, we found the Orchid Embryo extracts showed good potential on the antioxidative activity, therefore the CHCl3 layer of Orchid Embryo MeOH extracts was subjected to the constituent separation and screened the bioactivity. Twenty-two constituents, including six ionones, six benzenoids, two malate acid derivatives, one sterol, one lignin, four fatty acid analogs and one triacylglycerol, were isolated from the CHCl3 layer of Orchid Embryo extracts. Among them, nineteen compounds were isolated for the first time from Phalaenopsis species, and dimethyl-2-benzyl malate was the first reported from the nature source. Besides, twelve constituents were under investigation for the structure determinations.
In this study, the ethanol extract of Orchid Embryo was further fractionated by the normal and reverse phase processes. The extracts and fractions were evaluated for their radical scavenging activity (DPPH assay, phenol red-horseradish peroxidase assay), lipid peroxidation inhibition (estimation of conjugated dienes, thiobarbituric acid reactive substances assay, ferric thiocyanate assay, and ferrous ion chelating activity), tyrosinase inhibition activity, anti-collagenase activity and α-glucosidase inhibition activity. The results of the activity test indicated that the ethanol extract of Orchid Embryo and the further fractions showed significant activities on phenol red-horseradish peroxidase assay, inhibited conjugated dienes formation, and also thiobarbituric acid reactive substances assay. The Orchid Embryo extracts also showed good activities on the ferric thiocyanate assay, ferrous ion chelating assay, tyrosinase inhibition and anti-collagenase assay. Besides, the extracts also presented the potential towered the inhibition of α-glucosidase.
The present study points out the potential of Orchid Embryo extracting components and the further fractions as functional ingredients for cosmetic formulation and health supplements.

誌謝 Ⅰ
中文摘要 Ⅱ
Abstract Ⅲ
目錄 Ⅳ

第一章 緒論 1
第一節 蘭胚 1
第二節 研究背景與目的 2

第二章 文獻探討 3
第一節 蝴蝶蘭屬植物活性探討 3
第二節 蝴蝶蘭之成分探討 3

第三章 材料與方法 6
第一節 儀器與材料 6
第二節 活性測試 8

第四章 蘭胚之培養、萃取與分離 17
第一節 蘭胚之培養 17
第二節 蘭胚之萃取與分離 17

第五章 蘭胚化合物之構造研究 22
第一節Loliolide (9) 之構造研究 22
第二節Isololiolide (10) 之構造研究 27
第三節 3-Hydroxy-α-ionone (11) 之構造研究 31
第四節 Blumenol C (12) 之構造研究 33
第五節 3-oxo-α-Ionol (13) 之構造研究 35
第六節 Dehydrovomifoliol (14) 之構造研究 37
第七節 Dimethyl-2-benzyl malate (15) 之構造研究 39
第八節 Dimethyl-2-(4-hydroxybenzyl) malate (16) 之構造研究 41
第九節 Pinoresinol (17) 之構造研究 43
第十節 Methyl ferulate (18) 之構造研究 45
第十一節 Dihydroferulic acid methyl ester (19) 之構造研究 47
第十二節 4-Hydroxybenzyl alcohol (20) 之構造研究 49
第十三節 p-(Methoxymethyl) phenol (21) 之構造研究 51
第十四節 Methyl palmitate & Methyl linoleate (22) (23) 之構造研究 53
第十五節 其它化合物之構造研究 55

第六章 蘭胚萃取物之活性測試研究 57

第七章 結論 72

參考文獻 74
附錄一 光譜數據 77
附錄二 植物之萃取與分離 85

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