葛根中1个新的6a,11a-脱氢紫檀素

目的研究野葛Pueraria lobata根茎的化学成分。方法运用HP-20大孔吸附树脂、硅胶、ODS和Sephadex LH-20等柱色谱以及反相制备型HPLC等各种现代色谱分离技术进行系统的分离纯化,根据化合物的光谱数据和理化性质进行结构鉴定。结果从野葛根茎的甲醇提取物中分离得到11个化合物,分别鉴定为葛檀素(1)、芒柄花苷(2)、大豆苷(3)、大豆素(4)、4′,8-dimethoxy-7-O-β-D-glucopyranosylisoflavone(5)、异芒柄花苷(6)、染料木素(7)、formononetin-7-O-β-D-apiofuranosyl-(1→6)-O-β-D-glucopyranoside(8)、7,2′,4′-trihydroxyisoflavone(9)、3′-methoxydaidzein(10)和芒柄花素(11)。结论化合物1为1个新的6a,11a-脱氢紫檀素,化合物8和9为首次从葛属植物中分离得到。     

三种赣产葛地上部分总黄酮提取工艺优化及含量测定

目的:比较三种赣产葛根及其地上部分总黄酮的含量,优选葛地上部分总黄酮的最佳提取工艺条件。方法:以葛根素为对照品,采用紫外-可见分光光度法测定三种赣产葛的根、茎、叶和花中总黄酮的含量;并以乙醇浓度、料液比和提取时间为考察因素,通过正交试验确定葛地上部分总黄酮的最佳提取工艺。结果:不同基源的葛及其地上部分总黄酮含量差异较大,经正交试验得出最佳提取工艺为乙醇浓度60%、料液比1:50和回流提取1.5 h。结论:该含量测定方法操作简单、快速、准确,可用于同时测定葛不同部位的总黄酮含量;总黄酮提取工艺参数的优化将为葛地上部分资源的开发利用提供理论基础和科学依据。     

Profiling and quantification of isoflavone-C-glycosides impurities in puerarin injection by liquid chromatography coupled to ESI-ion trap mass spectrometry

An HPLC/DAD/MSⁿ method was established for the qualitative and quantitative analysis of the impurities in puerarin injection (PI), a widely used drug in China. The analytical HPLC was performed on an Agela RP-C18 column using 0.1% aqueous formic acid (v:v) and methanol as mobile phase. A total of nine impurities were detected and eight of them were identified as isoflavone-C-glycosides basing on their UV spectra and MSⁿ spectra and comparing with the literature data. An HPLC method for the assay of two common impurities in the commercial PI samples, i.e., neopuerarin A and neopuerarin B, was then established. The validation of the method, including sensitivity, linearity, precision, accuracy, was carried out. The calibration curves showed good linearity of R² > 0.9999 and LOQ (S/N = 10) were less than 3.73 ng. The precision was evaluated by intra- and inter-day assays and R.S.D. values were less than 0.94%. The average recovery rates were 97.0% and 99.5%, respectively, with R.S.D. less than 1.38%. The contents of neopuerarin A and neopuerarin B in various commercial brands of PI samples varied over the range of 0.30–1.16% and 0.42–1.66%, respectively. This is the first report on the impurities in PI.     

Acute Effect of High‐dose Isoflavones from Pueraria lobata (Willd.) Ohwi on Lipid and Bone Metabolism in Ovariectomized Mice

We investigated the acute metabolic effects of isoflavones from Pueraria lobata (Willd.) Ohwi (IPL) in ovariectomized (OVX) mice. After 4 weeks of IPL feeding at 500 mg/day/kg body weight (OVX500), plasma 17β‐estradiol concentrations were significantly higher (+25%, p < 0.05), whereas plasma triglyceride levels were significantly lower in OVX mice (?15%, p < 0.05) compared with controls. Abdominal adipose tissue weight was marginally reduced in IPL‐fed groups compared with OVX controls and the plasma levels of liver enzymes were unchanged. In addition, IPL significantly inhibited the reduction of bone mineral density in the femurs of OVX mice (OVX200, +22%; OVX500, +26%; p < 0.05) compared with controls after 4 weeks of IPL feeding. In quantitative polymerase chain reaction analysis the expression of aromatase was significantly suppressed and SULT1E1 was increased by IPL feeding, showing that IPL feeding may not alter the risk for breast cancer in mice. Our results suggest that IPL could ameliorate menopausal symptoms in mice. Further studies will confirm the effects of IPL in humans. Copyright © 2012 John Wiley & Sons, Ltd.     

罗布麻、葛根提取物对神经刺激所致猪脑动脉反应的影响

目的观察罗布麻、葛根提取物对神经刺激引起的猪脑动脉反应的影响及其与一氧化氮的关系。方法采用离体去除内皮血管方法,测定脑动脉螺旋条的等容张力变化及药物的作用。结果罗布麻、葛根提取物对电刺激或化学刺激引起的血管舒张反应具有增强作用,此作用不受酚妥拉明和哌唑嗪、心得安、阿托品、引哚美辛的影响,但可被一氧化氮合酶抑制剂L-硝基精氨酸抑制,该抑制作用又可为一氧化氮供体L-精氨酸逆转。结论罗布麻、葛根提取物对电刺激或化学刺激所致血管舒张反应的增强作用可能与一氧化氮有关。     

某种含葛根黄酮的保健食品的急性毒性和遗传毒性研究

[目的]研究某种含葛根黄酮的保健食品的急性毒性和遗传毒性,为葛根黄酮的开发利用提供依据。[方法]应用小鼠急性经口毒性试验、Ames试验、小鼠骨髓嗜多染红细胞微核试验和小鼠精子畸形试验对该含葛根黄酮的保健食品的毒性进行了试验观察。[结果]该含葛根黄酮的保健食品对雌雄小鼠LD50均大于10 000 mg/kg.bw,属实际无毒物质;Ames试验中受试物各剂量组回变菌落数均未超过自发回变菌落数的2倍,亦无剂量-反应关系,结果为阴性;小鼠骨髓嗜多染红细胞微核试验和小鼠精子畸形试验中各剂量组微核率、精子畸形率与阴性对照组之间差异无统计学意义(P>0.05),而环磷酰胺阳性对照组与阴性对照组之间差异有统计学意义(P<0.01),证明实验结果均为阴性。[结论]本次实验条件下,该含葛根黄酮的保健食品属实际无毒物质,未显示有遗传毒性作用。     

葛根提取工艺的考察

为优选葛根素的提取工艺 ,本实验采用 4因素 3水平正交设计法 ,以葛根素为指标 ,筛选影响葛根醇提工艺的因素。结果表明实验设计 4因素均具有显著性影响。结合生产实际需要 ,葛根最佳提取条件是 :加入 18倍量 80 %乙醇回流提取 3次 (8,6 ,4 ) ,提取 5h(2 ,2 ,1)。     

部分粉葛品种遗传关系的SRAP研究

目的:粉葛品种间遗传关系的分析.方法:以野葛和苦葛为外组,18个粉葛栽培品种为试材,通过SRAP分析,利用TREECONW软件分析遗传距离,UPGMA方法聚类,构建聚类树状图.结果:22对引物组合共得到338条扩增条带,其中有216条呈现多态性,占63.9%,平均每对引物组合产生15.4个位点和9.8个多态性位点,揭示了粉葛较为丰富的遗传多样性.粉葛品种遗传距离变化范围在0.0047～0.2658,平均0.136;聚类结果显示18个粉葛品种及外组被明显地划分三大聚类群,大部分品种并没有按地域形成独自的类群.结论:SRAP标记为粉葛遗传关系研究提供了新的DNA分子标记技术手段.     

葛根总黄酮分散片的药动学研究及其与愈风宁心片的比较

目的：研究葛根总黄酮分散片在大鼠体内的药代动力学特征及生物利用度,并与市售愈风宁心片进行比较。方法：大鼠灌胃给药葛根总黄酮分散片（受试制剂）及市售片（参比制剂）混悬液,采用HPLC测定给药后各时间点的血药浓度,用统计软件计算主要药代动力学参数。结果：分散片与市售片的Cmax分别为（20.21±0.18）,（9.90±0.25）mg·L^-1;分散片的Tmax=（0.50±0.03）h,市售片T_max=（1.09±0.02）h;分散片相对于市售片的生物利用度为182.68%。结论：葛根总黄酮分散片吸收快,起效时间明显提前,生物利用度明显优于市售片。     

酮通颗粒质量标准研究

目的:建立酮通颗粒的质量标准。方法:采用薄层色谱(TLC)法鉴别制剂中三七、银杏叶、葛根药材;用高效液相色谱(HPLC)法测定制剂中银杏总黄酮醇苷的含量。结果:TLC斑点清楚,专属性强。槲皮素、山柰素、异鼠李素的检测浓度分别在5.24～83.84、4.56～72.96、2.48～39.68μg·mL-1范围内与各自峰面积积分值呈良好线性关系(r分别为0.9992、0.9991、0.9994);三者平均回收率分别为96.21%、99.14%、95.00%,RSD分别为2.06%、1.19%、1.31%。结论:所建标准可用于酮通颗粒的质量控制。