【作者】 杨琳；

【导师】 孙启时； 车庆明；

【作者基本信息】 沈阳药科大学 ， 生药学， 2007， 博士

【摘要】 黄酮类化合物在植物界中分布广泛，为植物体内的重要次生代谢产物。研究表明，黄酮类化合物具有抗炎、抗氧化、抗自由基、抗衰老、抗病毒、抗癌、抗心血管疾病等多种药理活性。（1）本论文对甘草废渣中的黄酮类化学成分进行了研究，首次从甘草废渣95％乙醇提取物中分离并鉴定了6个黄酮类化合物，分别为甘草查尔酮A（licochalcone A，1）、甘草查尔酮B（licochalcone B，2）、刺甘草查尔酮（echinatin，3）、甘草黄酮（licoflavone，4）、7，4′-二羟基黄酮（7，4′-dihydroxyflavone，5）、5，7，4′-三羟基-8-异戊烯黄酮（5，7，4′-trihydroxy-8-isopenenylflavone，6）。（2）本论文首次建立了甘草废渣中甘草查尔酮A的HPLC含量测定方法，采用BISCHOFF C₁₈柱（2.0 mm×150 mm，5μm），甲醇-四氢呋喃-1％冰醋酸溶液（46：10：44）为流动相，流速为0.3 ml／min，检测波长为377 nm，柱温为30℃，结果表明甘草查尔酮A在1～32μg／ml范围内呈良好线性关系（r=0.9997），平均回收率为95.7％（RSD为3.0％）。并采用此方法对工业甘草废渣及甘草药材中的甘草查尔酮A的含量进行了比较研究。（3）本论文首次对甘草查尔酮A口服给药后血清中的代谢产物进行了研究。运用HPLC-DAD和LC-MS分析技术，对甘草查尔酮A在大鼠血清中的主要代谢产物进行了鉴定，分别为甘草查尔酮A-4-O-葡萄糖醛酸苷（licochalcone A 4-O-glucuronide）、甘草查尔酮A-4′-O-葡萄糖醛酸苷（licochalcone A 4′-O-glucuronide）及原形药甘草查尔酮A（licochalcone A）。（4）本论文对中药红花中的6-羟基山柰酚-3-O-葡萄糖苷进行了分离制备，首次对6-羟基山柰酚-3-O-葡萄糖苷灌胃给药后其在大鼠血清中的代谢产物进行了研究。建立了大鼠血清中6-羟基山柰酚-3-O-葡萄糖苷代谢产物的HPLC-DAD及LC-MS／MS分析方法，检测并鉴定了7个代谢产物，分别为6-羟基山柰酚-6，4′-二-O-葡萄糖醛酸苷（6-hydroxykaempferol 6，4′-di-O-glucuronide）、6-羟基山柰酚-3，6-二-O-葡萄糖醛酸苷（6-hydroxykaempferol 3，6-di-O-glucuronide）、6-羟基山柰酚-3-O-葡萄糖醛酸苷（6-hydroxykaempferol 3-O-glucuronide）、6-羟基山柰酚-4′-O-葡萄糖醛酸苷（6-hydroxykaempferol 4′-O-glucuronide）、6-羟基山柰酚-6-O-葡萄糖醛酸苷（6-hydroxykaempferol 6-O-glucuronide）、6-羟基-7-甲氧基山柰酚-3-O-葡萄糖醛酸苷（6-hydroxy-7-methoxyl-kaempferol 3-O-glucuronide）和6-羟基-3-甲氧基山柰酚-7-O-葡萄糖醛酸苷（6-hydroxy-3-methoxyl-kaempferol 7-O-glucuronide）。（5）本论文首次对灯盏花乙素苷元在大鼠血浆内的代谢产物进行了研究，建立了大鼠血浆中灯盏花乙素苷元及其代谢产物的HPLC-DAD及LC-MS／MS分析测定方法，采用该方法检测并鉴定了灯盏花乙素苷元在大鼠血浆中的4个代谢产物及原形药，分别为灯盏花乙素（scutellarin）、灯盏花乙素苷元-6-O-葡萄糖醛酸苷（scutellarein 6-O-glucuronide）、6-甲基灯盏花乙素（6-methoxyscutellarin）、灯盏花乙素苷元-4′-O-葡萄糖醛酸苷（scutellarein 4′-O-glucuronide）[或灯盏花乙素苷元-5-O-葡萄糖醛酸苷（scutellarein 5-O-glucuronide）]和灯盏花乙素苷元（scutellarein），初步阐明了灯盏花乙素苷元在大鼠体内的存在形式，为其作为口服制剂替代灯盏花乙素注射液提供了科学的依据。（6）本论文首次对口服给药后黄芩素在大鼠及人体内的代谢产物进行了研究，建立了人体血浆及大鼠血清中黄芩素和其代谢产物的HPLC-DAD及LC-MS／Ms分析测定方法。采用该方法从大鼠血清中检测并鉴定了5个代谢产物及原形药，分别为黄芩素-6，7-二-O-葡萄糖醛酸苷（baicalein 6，7-di-O-glucuronide）、黄芩素-7-O-葡萄糖醛酸苷（baicalein 7-O-glucuronide，即baicalin）、6-甲氧基黄芩素-7-O-葡萄糖醛酸苷（6-methoxybaicalein 7-O-glucuronide）、黄芩素-6-O-葡萄糖醛酸苷（baicalein 6-O-glucuronide）、黄芩素-7-O-葡萄糖醛酸甲酯（baicalein 7-O-glucuronyl methyl ester）和黄芩素（baicalein）；从人体血浆中检测并鉴定了5个代谢产物及原形药，分别为黄芩素-6，7-二-O-葡萄糖醛酸苷（baicalein 6，7-di-O-glucuronide）、黄芩素-7-O-葡萄糖醛酸苷（baicalein 7-O-glucuronide）、6-甲氧基黄芩素-7-O-葡萄糖醛酸苷（6-methoxybaicalein 7-O-glucuronide）、黄芩素-6-O-葡萄糖醛酸苷（baicalein 6-O-glucuronide）、7-甲氧基黄芩素-6-O-葡萄糖醛酸苷（7-methoxybaicalein 6-O-glucuronide）和黄芩素（baicalein）。口服给药后，黄芩素在大鼠体内及人体内均主要发生Ⅱ相代谢反应，代谢产物均以黄芩素-7-O-葡萄糖醛酸苷为主，但代谢产物不尽相同。更多还原

【Abstract】 Flavonoids is a kind of important secondary metabolites in plants and has varied activities such as anti-inflammatory, anti-free radical, anti-aging, antiviral, anti-cancer, anti-angiocardiopathy, et al.（1） In this paper, flavonoids in licorice residue was investigated, and 6 compounds （1～6） were isolated from the 95% ethanol extract of licorice residue by chromatographic methods. On the basis of spectroscopic analysis (¹H-NMR, ¹³C-NMR, EI-MS), they were identified as licochalcone A （1）, licochalcone B （2）, echinatin （3）, licoflavone （4）, 7, 4’-dihydroxyflavone （5）, and 5, 7, 4’-trihydroxy-8-isopenenylflavone （6）.（2） An HPLC-UV method was developed for the determination of licochalcone A in licorice residue and in licorice. The analysis was carried on the column of BISCHOFF C₁₈ （2.0 mm×150 mm, 5μm）, with the mobile phase consisted of methanol- tetrahydrofuran-1% acetic acid （46: 10: 44）. The flow rate was 0.3 ml/min with the detect wavelength at 377 nm, and the column temperature in 30℃. The method displayed excellent linearity in the ranges 1～32μg/ml with the regression coefficient at 0.9997. The average recovery of licochalcone A was 95.7% （RSD=3.0%）. With this method, the content of licochalcone A had been compared between licorice residue and licorice from different regions.（3） On the basis of the chemical study, the metabolites of licochalcone A in rat serum were investigated. With the estabolished methods of HPLC-DAD and LC-MS, 2 metabolites together with the parent drug were verified as licochalcone A 4-O-glucuronide, licochalcone A 4’-O-glucuronide, and licochalcone A.（4） 6-hydroxykaempferol 3-O-glucoside （7） was isolated and accumulated from Carthamus tinctorius L. And the metabolites of 6-hydroxykaempferol 3-O-glucoside in rat serum were investigated. Serum sample after oral administration of 6-hydroxykaempferol 3-O-glucoside was analyzed by HPLC-DAD and LC-MS/MS, and 7 metabolites were detected and identified as 6-hydroxykaempferol 6, 4’-di-O-glucuronide, 6-hydroxykaempferol 3,6-di-O-glucuronide, 6-hydroxykaempferol 3-O-glucuronide, 6-hydroxykaempferol 4’-O-glucuronide, 6-hydroxykaempferol 6-O-glucuronide, 6-hydroxy-7-methoxykaempferol 3-O-glucuronide, and 6-hydroxy-3-methoxykaempferol 7-O-glucuronide.（5） The HPLC-DAD and LC-MS/MS methods for determination of scutellarein and its metabolites in rat plasma were established. 4 metabolites together with the parent drug were detected and identified as scutellarin, scutellarein 6-O-glucuronide, 6-methoxyscutellarin, scutellarein 4’-O-glucuronide （or scutellarein 5-O-glucuronide）, and scutellarein.（6） Metabolites of baicalein after oral administration were investigated in rats and in humans, respectively. With the estabolished methods of HPLC-DAD and LC-MS/MS, 5 metabolites and together with the parent drug were detected from the rat serum and identified as baicalein 6, 7-di-O-glucuronide, baicalein 7-O-glucuronide （baicalin）, 6-methoxybaicalein 7-O-glucuronide, baicalein 6-O-glucuronide, baicalein 7-O-glucuronyl methyl ester, and baicalein. 5 metabolites and the parent drug were detected in the plasma of humans, and were identified as baicalein 6, 7-di-O-glucuronide, baicalein 7-O-glucuronide （baicalin）, 6-methoxybaicalein 7-O-glucuronide, baicalein 6-O-glucuronide, 7-methoxybaicalein 6-O-glucuronide, and baicalein. The major metabolite of baicalein both in rats and in human beings is baicalin, while the metabolites in human beings are not all the same as those in rats.更多还原