Radix Puerariae (Pueraria root, kudzu root) is the root of Pueraria lobata (Willd.) Ohwi or Pueraria thomsanii Benth. Radix Puerariae has been used as Chinese herb for a long time. The medical functions are as following: expels pathogenic factors from muscles and skin to reduce fever, promotes eruption, promotes the production of body fluids to quench thirst and invigorates the vital functions to relieve diarrhea. It has excellent effects cardiovascular system proved in pharmacology. To establish a rapid and automatic quality control method for Radix Puerariae, three major biological active components of isoflavone including daidzein, daidzin and puerarin in Radix Puerariae, were extracted by pressurized solvent extraction (PSE) and the conditions were optimized. Analysis of isoflavones were performed by using both high performance thin layer chromatography (HPTLC) and high performance liquid chromatography coupling with diode array detector (HPLC-DAD). 1. Analysis of isoflavons from Radix Puerariae using HPTLC HPTLC method for quantitative analysis of isoflavones from Radix Puerariae was developed. The results showed that it was not a good method for quantitative determination of the investigated isoflavones from Radix Puerariae because of its poor precision, reproducibility and resolution. 2. Analysis of isoflavone from Radix Puerariae using PSE and HPLC Sample preparation, the bottleneck of most analytical procedures, is the key step for quality control of TCM. One of the most promising and sample preparation techniques is the PSE (Dionex trade name ASE for accelerated solvent extraction), which offers the advantages of reducing solvent consumption and allowing for automated sample handling. Isoflavones in Radix Puerariae were extracted by using PSE. The operating parameters including extraction solvent, particle size, pressure, temperature and extraction times were optimized by using univariate approach in order to obtain the highest extraction efficiency. The optimized conditions are as follows: solvent, methanol; temperature, 140 °C; pressure, 1400psi and extraction time, 10 minutes. The results showed that PSE is a simple, efficient and rapid method with lower solvent consumption. HPLC-DAD was also developed for quantitation of isoflavone in Radix Puerariae. Simultaneous determination of puerarin, daidzein and daidzin was performed using HPLC-DAD and PSE. The results showed that HPLC-DAD was a good method for quantitative analysis of isoflavones in Puerariaroot. The separation was performed in an Agilent Eclipse XDB-C18(150 x 4.6mm, 3.5 u m) column with a mobile phase of methanol: 1 %HAC solution (gradient elution), the detection wavelength was 250 nm. Keywords: Pressurized solvent extraction; High performance liquid chromatography; High performance thin layer chromatography; puerarin; daidzein; daidzin; quality control.

葛根為豆科植物野葛 Pueraria lobata (Willd.) Ohwi 或甘葛藤 P.thomsonii Benth 的根,具有發表解肌、開陽透疹、解熱生津、升陽止瀉等作用,對心血管系統疾病有較好的治療作用。針對澳門現階段缺乏中藥質量控制的方案以及澳門自然、社會的條件,通過文獻調研,利用加速溶劑提取技術(PSE),考察 HPLC及高效薄層色譜(HPTLC)方法同時進行葛根中主要活性成份葛根素、大豆苷及大豆苷元含量測定,希望建立一個實現自動化和快速分析的葛根質量控制方法,為葛根的質量控制奠定基礎。論文實驗部分包括:1. 葛根異黃酮類成份高效薄層色譜分析採用加壓溶劑提取作為葛根樣品前處理手段,使用高效薄層板,半自動點樣,自動展開,光密度掃描,測定波長250mm,參比波長330mm,測定了葛根素、大豆苷、大豆苷元三種成分。結果表明:HPTLC 作為葛根質量控制的含量測定方法存在明顯不足,現有文獻與本研究均未能達到三種成分的完全分離;方法的重現性、精密度較低;影響因素多,難於滿足定量分析的要求。2.葛根異黃酮類成份加壓溶劑提取-高效液相色譜分析加壓溶劑提取是近年來發展起來的一種新的樣品處理技術,具有提取時間短、溶劑消耗少、提取效率高、重現性好、操作模式多樣化以及操作過程自動化等優點。以葛根素、大豆苷和大豆 元總量為評價指標,HPLC-DAD方法測定,用正交設計考察的方式,對葛根異黃酮類成分進行了加壓溶劑提取條件的優化。得到的優化條件為:選用甲醇作為ASE提取溶劑,提取溫度140℃,時間10分鐘,壓力1400psi,置換體積60%。定量分析最佳色譜條件:色譜柱Agilent Eclipse XDB-C18 (150 x 4.6mm, 3.5 μm),柱溫:25℃,流速1.0ml/min,進樣量51;甲醇-水(1%HAc)為流動相進行梯度洗脫;檢測波長為250mm;結果表明:加壓溶劑提取-高效液相分析方法快速;溶劑使用量少,利於環保;自動化程度高;適用於對葛根中葛根素、大豆苷及大豆苷元的同時定量測定。 關鍵字:加速溶劑提取技術;高效液相色譜法;高效薄層色譜法;葛根素;大豆苷;大豆苷元;葛根