牛樟芝培養及樟芝三萜類HPLC分析研究探討

Cultivation of Antrodia camphorata and Quantitation of Triterpenoids Using HPLC Analytical Method

江効儒(Shiau-ru Chiang)

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士班/碩士(2012年)

摘要

樟芝(Antrodia camphorata)，又名牛樟芝及樟芝，是台灣的一種藥用真菌，廣泛用於亞洲及歐洲國家的食品添加中，有預防癌症和保肝作用樟芝由其中空的牛樟樹幹內面長出，從樟芝萃取出的三?類化合物具有很好的生物活性，進行有抗癌及血管舒張之功效。因此本研究以樟芝三?類為指標，進行栽培條件及萃取方法之探討。研究中使用HPLC分析樟芝萃取出來的12種三?類化合物，分析時利用J’sphere ODS-M80 C18管柱，以0.1%甲酸及Acetonitrile作為移動相，流速為1 mL/min，偵測波長為248 nm，注射體積為5 μL，並以0.25-4 mg/mL的濃度製作11種三?類檢量線，探討在平面培養、液態培養及子實體中三?類成份的差異性。

關鍵字

三?類 ； 子實體 ； 液態培養 ； 平面培養 ； HPLC ； 牛樟芝

並列摘要

Antrodia camphorata (AC) (Syn: A. cinnamomea or Taiwanofungus camphorates), named “Niu-chang-chih” or “chang-chih” in Chinese, is a medicinal mushroom being widely used as food dietary supplement for cancer prevention and hepatoprotection in several Asian and European countries. AC is grown in inner cavity of the endemic species Cinnamomum kanehirae Hayata (Lauraceae). Previous studies on AC extracts as well as pure compounds revealed that it has huge range of bioactivities from anti-cancer to vasorelaxation. The chemistry of AC leads to the identification of more than hundred secondary metabolites. Among these, particular attention has been directed to triterpenoids which are reputed as the most potent that contribute to AC’s pharmacological efficacy. Therefore, triterpenoids are ideal marker for standardization of AC. The quantities of active triterpenoids in AC are mainly dependent on cultivation conditions, harvest period, and extraction methods. To evaluate the quality of AC, in this study, a simple and sensitive HPLC-UV separation method was developed for simultaneous determination of 12 triterpenoids in the extract of AC. Optimum separations were obtained with J’sphere ODS-M80 C18 column using gradient elution with 0.1% aqueous formic acid and acetonitrile as mobile phase, flow rate of 1 ml/min, detection wavelength at 248 nm and sample volume at 5 μL. This method produced linear responses for ten triterpenoids in the concentration ranges of 0.25–4 mg/ml. Our results showed that there were remarkable differences in triterpenoids contents from different AC samples such as culture plate, liquid culture, and fruiting bodies, etc.

並列關鍵字

triterpenoids ； fruiting bodies ； liquid culture ； culture plate ； HPLC ； Antrodia camphorata

參考文獻

林苡芬 「不同發酵碳源之牛樟芝菌絲體發酵過濾液對人類肝癌細胞株之影響」。國立臺灣大學食品科技研究所碩士論 (2004)。

Heng-Hua Wu, Zhi-He Yu, Yu-Cheng Dai.(2004) Taiwanofungus, a polypore new genus .Fung. Sci. 19(3–4): 109–116.

張東柱 「靈芝與熱帶靈芝子實體型態與菌絲純培養之比較」。林業試驗所研究報告季刊，6, 397-406 (1991)。

高曉薇 「臺灣靈芝屬新種樟芝之三?類成分研究」。臺北醫學院天然物醫物研究所碩士論文 (1992)。

Chang, T. T. &; W. N. Chou,(1995) Antrodia cinnamomea sp. nov. on Cinnamomum kanehirai in Taiwan. Mycol. Res. 99(6):756-758.。