高效液相層析紫外光偵測器飛行式質譜儀於牛樟芝子實體乙醇萃取物之分析

Analysis of Ethanolic Extracts of Antrodia Cinnamomea by High-Performance Liquid Chromatography / Ultraviolet Detector/ Time of Flight Mass Spectrometry

林信億(SIN-YI Lin)

指導教授 ： 沈振峯

國立虎尾科技大學/文理學院/生物科技研究所/碩士(2015年)

摘要

牛樟芝（Antrodia cinnamomea）為台灣的一種珍貴之食藥用真菌類，具有保肝、抗癌等功效。許多科學研究已經證明，牛樟芝的子實體提取物具有多種藥理作用和生物活性例如:抑制B型肝炎病毒、抑制癌細胞生長等，因此廣泛被用在保健食品的開發，傳統上，牛樟子實體的指標成分主要是使用LC-UV進行分析，因此，我們參考之前文獻所提出分析13個指標化合物的方法，建立出我們的牛樟芝子實體抽出物之化學指紋圖譜，且本研究引入質譜偵測，增加了分子離子及離子碎片的資訊，強化定性分析的可信度。同時，透過置換層析管柱和層析參數的調整，我們使分析時間大幅縮短為20分鐘。我們利用這套系統分析各種牛樟菌樣品以探討其不同生長期及有無中藥處理之成分變化，結果顯示，經細辛處理過的牛樟芝仍含有豐富的指標成分，且經細辛處理過的牛樟芝型態發育會比未經細辛處理良好，因此更具有經濟價值，觀察不同生長期的成分變化，在6個月的樣品中以lanostane-type triterpenoids的含量最多；在12、14個月的樣品中則以ergostane-type triterpenoids 的較多，這和文獻上所觀測到的結果是一致的。我們亦測試了在不同偵測模式UV、ES+、ES-、DA+、DA-下指標物的訊號差異，在13個指標化合物中LC-MS能全檢測到，其中4個為含量較少之化合物，但可在LC-MS中清楚的觀測到，在LC-ES(+)-MS 及LC-DA(+)-MS中可觀測得最多化合物18個，在LC-MS和LC-UV均測的到的化合物中，LC-MS測的到的化合物均有一些帶電基團，且LC-MS訊號靈敏度皆明顯較LC-UV強，最後，我們也嘗試以熊果酸(ursolic acid)進行牛樟成分的半定量分析。本研究已初步利用LC-UV-TOF為牛樟芝萃取物建立一套快速、可靠之成分展開檢驗技術。

關鍵字

牛樟芝 ； 子實體 ； 液相層析飛行式質譜儀

並列摘要

Antrodia cinnamomea is a precious edible fungus endemic to Taiwan that has efficacy of hepatoprotective and anticancer. Numerous bioactivity studies have investigated and validation, Antrodia cinnamomea has extensive biological activities, such as anti-hepatitis B virus effects, anticancer activity et al. Traditionally, the major marker compounds of ruiting bodies of Antrodia cinnamomea were analyzed with LC-UV. Ting-Yu Lin et al. proposed a LC-UV method for 13 marker compounds which needed 120 minutes run time. In this study, the conditions of LC were improved to achieve a faster analytical method (20 min)for the profiling of the fruiting bodies of Antrodia cinnamomea. The mass spectrometric detection was also applied in this study to obtain the molecular ions and ion fragments information to reinforce the credibility in the qualitative analysis. The developed method was used to analyze various Antrodia cinnamomea related samples with or without the treatment of the Herb (Asarum). The results showed that the Asarum processed Antrodia cinnamomea still contains abundant marker components. For six months growth period samples, the lanostane-type triterpenoids were dominated ; for samples of 12 and 14 months growth, the ergostane-type triterpenoids were dominated. The composition change is consisted with the results reported by Ting-Yu Lin et al. The extracts were analyzed by different detection modes, i.e. UV, ES +, ES-, DA + and DA-. The results shown that the 13 marker compounds were all clearly detected with the LC-MS, in which 4 contents of the compounds are lese aboundent in the fruiting body. The LC-ES (+)-MS and LC-DA (+)-MS can observe the most markers (18 compounds). The signal sensitivity of LC-MS were significantly stronger than the LC-UV. Finally, we also try to develope a semi-quantitative analysis based on ursolic acid equivalent. This study has builded a fast, reliable compound profiling and semi-qutitative method for the Antrodia cinnamomea extracts by LC-UV-TOF-MS.

並列關鍵字

Antrodia cinnamomea ； fruiting body ； Liquid Chromatography Time-of-Flight Mass Spectrometry

參考文獻

參考文獻

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