The root and rhizome of Salvia miltiorrhiza Bge. (Danshen in Chinese) is one of valuable and commonly used traditional Chinese medicines (TCMs). It is known for its efficacy in promoting blood circulation, removing blood stasis, inducing blood clotting, relieving swelling, and alleviating pain. It has been widely used for treatment of cardio- and cerebro-vascular diseases. A lot of studies showed that some components of Salvia have the effect on anti-platelet aggregation and anti-thrombosis. To date, several methods, including HPTLC, HPLC, LC-MS, UPLC, CE and CEC, have been used for qualitative and/or quantitative analysis of chemical components in Danshen, but few analytes were determined in these papers. In addition, there is few report on chemical characteristics and markers for discrimination and quality control of Danshen produced and used in China. Modern pharmacological studies have shown that combining with some receptors or channels on cell membrane is the first step of drug action. Therefore, one method named platelet biospecific extraction has been established to find out the effective chemicals from TCMs. If the extract from TCMs had the effect on antiplatelet aggregation, the effective chemicals in it should be binding to the receptors or the other targets on membrane of platelets. After washing out the non-binding substance, the effective chemicals in the herbs were isolated from the receptors or the other targets on membrane. The analysis of the effective chemicals in the herbs can be conducted via chromatography such as HPLC. This method has been confirmed to be available and convenient. In this study, a UPLC method was developed to analyze 13 components in Salvia from different locations and in different parts of Salvia, and five components were found out, which suggested to be the appropriate markers IV 澳門大學碩士學位論文 for quality assessment of S. miltiorrhiza. And by human platelet biospecific extraction and this established UPLC method for screening potential antiplatelet aggregation active components in Danshen was also developed to optimize the appropriate markers for quality control of Salvia. The major contents of this study: 1. A reliable UPLC method had been developed to analyze 13 components in Salvia. Using this method, 74 samples of Salvia collected from different locations in China were analyzed and the chemical characteristics for different parts of Danshen were also investigated. And the contents of the investigated compounds for the roots, rhizome and aerial parts were also compared. The result showed that five components-salvianolic acid B, rosmarinic acid, crytotanshinone, tanshinone I and IIA may be the appropriate markers for quality assessment of S. miltiorrhiza. 2. A method, by human platelet biospecific extraction and UPLC for screening potential antiplatelet aggregation active components in Danshen, was developed to optimize the markers for quality control of Salvia. Four phenolic acids screened from water extraction of S. miltiorrhiza were danshensu, protocatechuic acid, protocatechualdehyde and salvianolic acid B, and 3 compounds of tanshinones were also screened in water extraction of S. miltiorrhiza. These ingredients in Salvia have been reported to be the antiplatelet aggregation active components in S. miltiorrhiza in the related papers. Four compounds, namely salvianolic acid B, rosmarinic acid, crytotanshinone, and IIA were considered to be the markers for quality assessment of Salvia. Keywords: Salvia miltiorrhiza Bge.; UPLC; Antiplatelet aggregation; Human platelet biospecific extraction

丹參為屑形科植物 Salvia miltiorrhiza Bge.的乾燥根及根莖,為我國傳統的 重要中藥之一,具有祛瘀止痛、活血通經及清心除煩等功效,近年來多用於治 療冠心病、心絞痛等心腦血管系統疾病。很多研究表明,丹參中一些成分具有 抗血小板聚集作用,抗血栓形成的作用。目前許多丹參的定性、定量分析方法 如HPTLC,HPLC,LC-MS,UPLC, CE 和CEC 已經建立,但由于測定的成 分或樣品量有限,不能全面反映丹參的化學特徵,而對于其質量評價指標的選 擇也尚待進一步研究。 中藥具有抗血小板聚集的作用,其中的活性成分應該能夠與血小板上的靶 點結合。基於此建立的血小板生物特異性萃取法已成為篩選中藥中相關活性成 分的有效、簡便的方法之一。 本實驗的主要內容為建立了丹參的超高效液相色譜分析方法,運用該方法 對收集到的丹參樣品進行分析,篩選出了可以作為其質量評價指標的有關成 分,並採用人血小板生物特異性萃取結合超高效液相色譜的分析方法對丹參水 提物中可能具有抗血小板聚集作用的成分進行了篩選,為丹參的質量評價指標 的進一步優化提供依據。 研究的主要內容包括: 1. 建立丹參 UPLC 分析方法,運用該方法對收集自我國各地74個月參或甘西 鼠尾草樣品以及丹參不同部位的成分進行了分析。通過對結果的聚類分析及指 紋圖譜分析評價了丹參樣品的化學特徵,比較了丹參根、根莖以及地上部分成 分含量的差异,並篩選出了5個可以作為丹參質控指標的成份一迷迭香酸、丹 酚酸B、隱丹參酮、丹參酮I和丹參酮 IIA。 2. 採用人血小板生物特異性萃取結合所建立的超高效液相色譜分析方法,篩選 出了丹參水提物中能與血小板特異性結合的成份一丹參素、原兒茶酸、原兒茶 醛、丹酚酸B、二氫參酮I、隱丹參酮以及丹參酮 IIA,已有文獻報道這些成 分都具有抗血小板聚集作用;結合化學成分分析與人血小板萃取實驗的結果, 篩選出四個成分一迷迭香酸、丹酚酸B、隱丹參酮和丹參酮 IIA,以這四個成分 Π 澳門大學碩士學位論文 的含量為指標對丹參及甘西鼠尾草樣品再一次進行了聚類分析,結果提示迷迭 香酸、丹酚酸B、隱丹參酮、和丹參酮 IIA 可以作為丹參的質量評價指標。 關鍵字:丹參;質量控制指標;超高效液相色譜;抗血小板聚集;人血小板生 物特異性萃取