澳門大學碩士論文 Abstract Chemical standards are the key substances for the qualitative and quantitative analysis, which play a key role for controlling the quality of traditional Chinese medicines (TCM). However, the chemical components especially the volatile compounds from TCM are not only difficult to obtain, but also hard to be preserved due to their unstability. Therefore, absence of chemical standards has become a bottleneck for the internationalization of Chinese medicines. The purpose of present study was focused on the development of digital chemical standard library for qualitative and quantative analysis of volatile componds in TCMs. Ezhu, one the commonly used Chinese herbal medicine, was selected as the target. Based on the main volatile components separated from Ezhu and a series of n-alkane, a digital chemical standard library was established, which can be used for their qualitative and quantitative analysis when there were absence of these volatile chemicals. There are four chapters for the thesis. Chapter I introduced the background and strategy for the present study. As the short of supply of chemical standards on the market, it is urgent to seek for alternative pathway without reference compound for quality control of Chinese medicines. Chapter II was for the establishment of digital chemical standard library. Firstly, the collection of MS data for the target compounds was obtained by using the Automatic Mass Spectral Deconvolution and Identification System (AMDIS). There were more than 40 compounds' MS data collected in the MS library. Secondly, GC-MS method was developed for the analysis of Ezhu spiked with a series of n-alkane. And then the relative retention times to adjacent alkanes as well as the MS data of the compounds can be used for the identification (qualitative analysis) when there were short of chemical standards. Finally, a quantitative method based on corrected factor was developed by using the chemical standards and n-alkanes, while the qualitative and quantitative validity for the developed method were tested. The results showed that the developed digital chemical standard library is feasible for the qualitative and quantitative analysis of the nine volatile components without or with a reference substance. Furthermore, the characteristics of the developed method were also compared with multi-components quantitation using one marker and chemical analogue methods. Chapter III was the application of the established digital chemical standard library for the analysis of 19 batchs of Ezhu samples collected from different hospitals distributed in China. While the quantitative result was compare to that of classic standard curve method; the results indicated that the virtual standard was practical and feasible for quality control of Chinese medicine. In a word, digital chemical standards were developed and employed for quality control of Chinese medicines, which is helpful to control the quality control of Chinese medicines when there are lack of reference substances. Key words: Digital chemical standard, volatile components, Chinese medicine, qualitative analysis, quantitative analysis, Ezhu V

中藥化學對照品是中藥定性定量研究的物質基礎,是中藥品質控制的關鍵。 然而由于中藥化學對照品特別是揮發性成分對照品存在難以獲得不易保存的缺 點,從很大程度上影響了中藥揮發性成分化學對照品的種類及數量,造成了中藥 品質控制研究及日常檢測所需化學對照品的嚴重不足,阻礙了中藥品質控制研究 的發展,成為中藥國際化的瓶頸。本研究針對中藥揮發性化學對照品缺乏的問 題,以常用中藥材莪術為研究對象,應用系列正構烷煙及實物對照品構建虛擬對 照品庫,并采用建立的虛擬對照品對其主要成分進行定性鑒別和定量測定研究, 從而探討建立中藥揮發性成分虛擬(數字)化對照品庫的可行性。 本論文包括三個章節。第一章介紹本課題的研究背景并提出本文的研究方 法通過調研中藥化學對照品的供求現狀并參照目前替代對照品方法的研究進展 分析了建立中藥揮發性成分數字化對照品的必要性及可行性。 第二章重點介紹了中藥(以莪術為實例)揮發性成分數字對照品庫的構建。 第一、二節為中藥揮發性成分化學對照品質譜數據的獲取:一方面通過化學分離 得到薑黃揮發油中的主要揮發性成分包括芳-薑黃烯、芳-薑黃酮、薑黃酮、β薑黃酮;另一方面通過購買市售的對照品得到39個中藥揮發性成分,并對這些 揮發性成分對照品進行 GC-MS 分析,獲得其MS 數據,應用 AMDIS 系統建立 MS 圖庫。第三節建立了我術主要揮發性成分(β-欖香烯、莪術烯、异莪術烯醇、 吉馬酮、莪術二酮、莪術烯醇、新莪術二酮及莪術雙環烯酮等9個成分)及其系 列相鄰正構烷煙的GC-MS 分析方法,并進行了方法學考察。第四節為對照品定 性參數的虛擬化 通過計算有效成分對照品及其相鄰正構烷煙的校正保留時間的 比值作為定性因子,求得各成分定性因子的變异係數 RSD%均小于0.05%,表明 運用該定性因子結合相應的MS 數據可以作為揮發性成分定性鑒別的有效方 法,達到了對照品定性參數虛擬化的目的。本章第五節主要為揮發性成分對照品 定量參數的虛擬化 首先采用對照品及其相鄰正構烷煙的響應值的比值建立定量 因子井驗證其準確度,但由于MS 的響應具有一定的波動性,此定量因子在實際 應用時需通過校正方可作為各化合物定量計算的依據。本文選取表術中常見成分 吉馬酮及其相鄰正構烷 Cin 的校正曲綫計算 MS 響應值校正因子,計算各化合 物的校正定量因子井進行二次驗證。驗證結果顯示,在一定濃度範圍內,以虛擬 對照品進行定量的結果與各對照品的實際濃度基本一致說明該方法在沒有對照 品的情况下對莪術中揮發性成分進行定量分析是可行的。同時,本節中將本文的 虛擬對照品法與文獻的一測多評法、類似物法進行了比較。 第三章是本研究在實際應用中的體現,用數字化對照品法對 19 批市售莪術 藥材進行了定性定量分析,同時與經典的標準曲綫法的定量結果進行比較,結果 基本一致,說明瞭本方法的實用性和可行性,體現了本研究的實際意義。 總之,本文針對對照品缺乏的問題,首次運用系列正構烷煙建立數字化對照 品的構想,以常用中藥莪術為例進行了初步研究,為中藥替代對照品研究提供了 有益參考。 關鍵詞:數字化對照品,中藥揮發性成分,定性分析,定量分析,我術 III