牛樟芝固態培養與應用AMP為內在標準品之三萜類HPLC分析研究探討

Solid state cultivation of Antrodia camphorate and HPLC analysis of triterpenoids using AMP as internal standard.

王楷敬(Kai-Ching Wang)

指導教授 ： 曾耀銘

朝陽科技大學/理工學院/應用化學系生化科技碩博士班/碩士(2015年)

摘要

三萜類(triterpenes)在自然界分佈廣泛，部分的植物、動物及菌類都含有此類化合物，但從不同動植物身上所發現的三萜類種類及含量又有些許不同。三萜類具有抑制癌細胞生長和抗氧化活性的特性，而一般的三萜類多是由羊毛甾醇(lanostane)或麥角甾醇(ergostane)所組成，則牛樟芝子實體具較高藥理活性之三萜類大都以麥角甾醇為主。本實驗利用固態基質培養牛樟芝菌絲，以液態培養形式，將菌液接種至固態培養基當中，觀察紀錄菌絲生長型態，以利進行三萜類含量分析。且本實驗擬測試運用單磷酸腺苷(AMP)做為內在標準品以HPLC定量分析牛樟芝三萜類，初步實驗證實AMP與三萜類可以相同的溶劑沖提，且兩者在HPLC當中有著不同的滯留時間，所以可進一步研究AMP是否適合做為三萜類HPLC定量分析的內在標準品化合物，適當發展之後從HPLC分析圖譜的波鋒面積比值計算與三萜類濃度之間的相關性，依其良好的相關性即可建立標準曲線並由標準曲線可推算出每一種樟芝樣品溶液中所含三萜類的濃度。

關鍵字

單磷酸腺苷 ； 牛樟芝 ； 三萜類

並列摘要

Triterpenoids are widely distributed in nature, some of the plants, animals and fungi contain these compounds, but the type and content of triterpenoids found in different plants and animals who have a little different, triterpenoids inhibit cancer cell growth and anti-oxidation activity characteristics.And general triterpenoids mostly by the lanostane or ergostane composed, the fruiting bodies of Antrodia with triterpenoids mostly in higher pharmacological activity of ergosterol-based.In this study, the use of solid matrix culture Antrodia mycelium in liquid culture in the form of the bacteria were inoculated solid medium, observation records mycelial growth patterns, in order to facilitate analysis of triterpenoids content.And experiments intended to test the use of adenosine monophosphate (AMP) as the internal standard in HPLC quantitative analysis of Antrodia triterpenoids, preliminary experiments confirmed that AMP and triterpenoids elution use the same solvent, and both of them have different in HPLC retention time. Therefore, further studies can be used as the internal standard compound triterpenoids HPLC. AMP is suitable for quantitative analysis, after proper development of the ratio calculated from the HPLC chromatogram of the area of the wave front triterpenes correlation between concentration, according to their good correlation resistance to the establishment of a standard curve can be calculated by the standard curve concentration of each sample solution contained Antrodia triterpenoids.

並列關鍵字

Antrodia ； Triterpenoids ； AMP

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