丹參有效成份快速分離及大量製備之流程探討

Preparation and analysis of bio-active components from Salvia miltiorrhiza Bunge

吳建德(Chien-Te Wu)

指導教授 ： 曾耀銘 ； 詹効松

朝陽科技大學/理工學院/生物技術與化學工程研究所碩士班/碩士(2010年)

摘要

丹參（Salvia miltiorrhiza Bunge）又名血參根、大紅袍、紅根赤參、夏丹參、紫丹參等，為唇形科（Labiatae）、鼠尾草屬（Salvia）植物，是著名的「活血化瘀」中藥，於「神農本草經」記載丹參列為上品，謂味苦、微寒。並有通經止痛、消炎等作用。近年來，於醫學上證實可保護心臟組織缺氧、血管舒張、抑制血小板凝集、抑制血栓形成等作用。
丹參中有效成份在醫療上已有很深入的研究，尤其以丹參酚酸B（Salvianolic acid B）對心血管疾病有著顯著的療效。本研究將針對丹參其有效成份丹參酚酸B，以樹脂HP20進行快速分離和大量製備進行探討，實驗將主要區分為製備樣品、吸附、脫附和再生四大程序。製備樣品程序，將探討乙醇對脂溶性丹參酮類化合物之提取效率，以及不同乙醇濃度對丹參酚酸B之萃取效率；吸附程序將探討吸附間隙流速（Interstitial velocity），以及進料濃度對樹脂吸附效率之影響；脫附程序將探討脫附間隙流速、不同脫附劑對丹參酚酸B脫附效率和回收效率及相對含量之影響；再生程序中將探討樹脂再生條件、包括再生溶劑的選擇和再生溶液體積對樹脂進行再生後吸附效率之影響。綜合以上所有最佳條件，本研究利用95 %乙醇萃取丹參藥材中之總丹參酮含量為0.05 %（0.52 mg/g），以60 %乙醇溶劑比例混合液於溶質溶劑比1：30（w：v），攪拌時間60分鐘，丹參酚酸B之提取效率可達93.8 %（42.68 mg/g），經HPLC分析初萃取丹參酚酸溶液之相對含量為8.75 %；在吸附程序中，丹參酚酸B於進料濃度4500 ppm，以吸附間隙流速為0.33 cm/s時有最高之總吸附量，每克樹脂之吸附量為8.24 mg；於脫附程序中，變換不同脫附劑與不同操作間隙流速之試驗結果，以95 %乙醇作為脫附劑於脫附間隙流速為0.14 cm/s，每克樹脂之脫附量為6.34 mg，其單管（40 mL收集一管）最高相對含量為77.2 %，管柱樹脂吸附後經由脫附程序其回收率達76.94 %；再生程序中，以70 %乙醇溶液添加4 %NaOH作為再生溶劑（1000 mL）進行再生，繼以三倍體積甲醇進行洗滌，經再生後以上述最佳操作條件重複吸附及脫附程序，於每克樹脂其總吸附量為7.81 mg和吸附率可達94.84 %。
經由萃取系統程序處理後，其丹參酚酸B從相對含量由8.75 %提升至77.2 %。為了測試其樹脂有效利用性，將樹脂經由吸附、脫附和再生程序重複三次後，其每克樹脂之吸附量均可維持在7.5~8.4 g；經由以上本試驗所研發之萃取程序，可將丹參中水溶性之主要成份丹參酚酸B化合物製備純化；本試驗於小規模萃取程序系統中進行試驗，將所得之參數於未來能將系統放大。

關鍵字

樹脂 ； 丹參酚酸B ； 丹參 ； 丹參酮類化合物 ； 間隙流速 ； 相對含量

並列摘要

Salvia miltiorrhiza Bunge (Labiatae) is a traditional Chinese medicinal herb. The dried roots of S. miltiorrhiza also called as Dan-shen is commonly used either on its own or in combination with other herbs for the prevention of cardiovascular diseases based on the concepts of traditional Chinese medicine. Recent Western medical techniques are successfully for specific actions of this herb on the cardiovascular system, such as inhibition of cellular adhesion molecule expression and adherence of inflammatory cells on vascular endothelium, attenuation of LDL oxidation and lipid actions, anti-platelet aggregation, and reducing endothelin release and improving local circulation. In this study, an improved and efficient process for the purification of cardio-protective compound, salvianolic acid B (SAB) from S. miltiorrhiza roots was developed. Starting with 95 % ethanol extraction of S. miltiorrhiza roots that contain 0.05 % of fat soluble tanshinones, it was possible to remove the first fraction enriched with tanshinones. The second extraction was producted by 60 % ethanol (1:30 w/v) and the extraction which contains 1423 ppm of SAB and the extraction rate was 93.8 %. The second fraction was subsequently introduced into a macroporous adsorption resins for recovering the SAB. The adsorption, desorption and regeneration conditions of the Diaion HP20 resin were carried out and compared, providing the high adsorption and desorption operational patrameters for SAB based on the research results. Dynamic adsorption and desorption was performed on Diaion HP-20 resin packed in a glass column (L 68 cm × OD 2.59 cm, with 100 g resin) to obtain optimal parameters for the separation of SAB. The optimum conditions of adsorption, desorption and regeneration processes were as below. In the 0.33 cm/s interstitial velocity and 4500 ppm SAB in the adsorption feeding exhibited the maximum adsorption of SAB (8.24 mg/g resin). In the desorption process, by using 95 % ethanol in the 0.14 cm/s interstitial velocity, 6.34 mg SAB per gram resin and 77.2 % relative content of SAB were attained. For resin regeneration, 1 litter 70 % ethanol which contains 4 % NaOH were used and then treated 3 BV of MeOH. Through the process, finally the reactive content of SAB was increased from 8.75 % to 77.25 %. This process were repealed three times and the the adsoption amounts of SAB remained stably between 7.5 to 8.4 mg / g. In this study, we have developed an efficient purification method for SAB and the content of SAB has also been increased. This purification parameters and operation conditions can be applied in large scale for the production of SAB.

並列關鍵字

Salvia miltiorrhiza ； Dan-shen ； salvianolic acid B (SAB) ； tanshinones ； Diaion HP20 ； Interstitial velocity

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