Ultrasonically Assisted Extraction of Isoflavones from Stem of Pueraria lobata （Willd.） Ohwi and Its Mathematical Model

Ultrasonically assisted extraction of isoflavones from the stem of Pueraria lobata （Willd.） Ohwi has been carried out with an ultrasonic extracting apparatus （20kHz, electrical power input to the transducer in 0-650W）. The influence of the electrical power input and extraction time on the＇extraction yield is investigated in water, n-butanol, and 95% （by volume） and 50% （by volume） ethanol aqueous solution. The experimental results indicate that the yields of total isoflavones are higher in ultrasonically assisted extraction than those obtained from con-ventional extraction.Moreover,a mathematical model is proposed,by introducing the electrical power input to index the ultrsound intensity,to describe the behavior of ultrasonically assisted extraction.It is found that the model calcuations are in good agreement with the experimental data.

Extraction Technology of Total Flavonoids from Pueraria lobata (Willd.) Ohwi and Content Determination of Pueraria lobata (Willd.) Ohwi at Different Altitudes

[Objectives]To optimize the extraction technology of total flavonoids from Pueraria lobata( Willd.) Ohwi. [Methods]The ultraviolet-visible spectrophotometry was used to determine the content of total flavonoids in Pueraria lobata( Willd.) Ohwi. With the puerarin as index,the reflux extraction and single factor test were employed to investigate the effects of temperature,time,ethanol concentration and solid-liquid ratio on the content of total flavonoids in Pueraria lobata( Willd.) Ohwi,respectively. Under the optimal extraction technology,the content of total flavonoids in Pueraria lobata( Willd.) Ohwi at different altitudes was determined.[Results] The optimum extraction process was as follows: 70%ethanol; solid-liquid ratio of 1∶ 30; 1 h reflux extraction. Under these conditions,the extraction rate of flavonoids in Pueraria lobata( Willd.) Ohwi was 11. 48%,the total flavonoids content of different kudzu parts was in the order of roots > stems > leaves,and the total flavonoids content of the sample at about an altitude of 1000 m was significantly higher than at the altitudes of 1400 m and 1700 m.[Conclusions]It was suggested that the Pueraria lobata( Willd.) Ohwi should not be cultivated as medicinal plant in too high mountains,and the stems and leaves of Pueraria lobata( Willd.) Ohwi could be used as raw materials for extracting total flavonoids.

基于HPLC指纹图谱结合一测多评法的葛花质量控制研究

建立葛花药材的指纹图谱,并同时建立其中10个主要黄酮类成分含量的一测多评分析方法(QAMS),用于不同产地葛花药材整体质量评价。采用高效液相色谱法(HPLC)对不同产地葛花中主要化学成分进行分析,确定最佳色谱条件,建立指纹图谱;在此基础上,以葛花苷为参照峰,建立QAMS测定葛花10个黄酮类成分含量;并与外标法(ESM)测定结果比较,验证QAMS的准确性。建立了葛花药材指纹图谱,20批葛花和10批粉葛花药材的相似度均大于0.85,标定共有峰分别为18和27个。以葛花苷为参照峰,各个成分校正因子的重复性、耐用性均良好;应用一测多评法测定10个成分的含量,与外标法测定结果比较,含量无明显差异(P>0.05),相对偏差<3.0%。不同产地来源的葛花药材中10个成分含量均存在较大差异,说明应提高葛花药材的质量控制标准。所建立的指纹图谱结合一测多评分析方法简便、可行,可客观、有效的用于不同产地葛花药材质量评价,可以为葛花质量标准修订、优质葛花种质资源的选育和育种提供参考。

葛根、粉葛炮制前后多糖和微量元素含量研究

目的:探讨不同的切制规格及炮制方法对葛根、粉葛所含多糖、微量元素含量的影响。方法:利用UV以及ICP-MS检测方法,对葛的不同品种(葛根、粉葛)、不同规格(片、丁)、不同炮制方法(煨制、醋制)中所含多糖和微量元素含量进行测定。结果:多糖类含量粉葛远高于葛根;丁块型葛基本高于片型葛;葛根丁经过煨制、醋制后没有显著变化,葛根片煨制提高、醋制降低,粉葛丁经过煨制、醋制后提高,粉葛片经煨制、醋制后降低。微量元素含量,葛根中Fe、Cu、Zn、Mg、K、Mn含量均高于粉葛,但Ca含量明显低于粉葛;Fe的含量均是片大于丁,Zn、Ca含量则是丁大于片;葛根丁中微量元素含量经过煨制后变化较大、醋制变化较小,葛根片中Fe、Cu、Zn、Mg、Mn含量煨制降低、醋制提高,粉葛丁中Fe、Zn、Mg、K、Ca含量经过煨制、醋制后降低,粉葛片中Fe、Zn、Ca含量经过煨制、醋制后降低。结论:葛根、粉葛不同的切制规格及炮制方法对其多糖和微量元素含量均可产生不同程度的影响,而该变化可为进一步深入探讨葛的炮制作用机理提供参考依据。

中药葛根防治糖尿病的作用机制和研究进展

糖尿病患病率高且并发症多,严重影响各种疾病的预后。在治疗上,降糖西药不良反应较多,限制临床治疗。中医药对于防治糖尿病经验丰富,可通过多靶点多通路降低患者血糖。中药葛根作为辛凉解表药,有清热、生津、止渴之效,药理学研究其具有抗氧化应激反应、减少炎症因子积聚、增强胰岛素敏感性、减少胰岛素抵抗等作用。葛根治疗糖尿病的主要活性成分为葛根素,临床研究表明以葛根素、葛根药对、葛根类方辨证论治各种糖尿病患者,疗效显著。近年来研究发现葛根素、葛根药对及葛根类方可以通过多靶点、多通路抑制氧化应激反应、调节体内炎症因子水平、减少AGEs的形成与累积、抑制胰岛β细胞凋亡、增加胰岛素敏感性、减少胰岛素抵抗等多种途径达到降低血糖,控制并改善各种并发症。文章旨在通过梳理相关文献,从葛根素、葛根药对、葛根类方3个方面总结归纳中药葛根防治糖尿病的作用机制和研究进展。

基于网络药理学探讨丹参-葛根药对治疗心肌纤维化的作用机制

目的采用网络药理学探讨丹参-葛根药对治疗心肌纤维化的活性成分及作用机制。方法从中药系统药理学分析平台获取丹参-葛根药对活性成分和作用靶点,通过GeneCards数据库获取心肌纤维化的相关靶点,使用Venny 2.1软件获取两者共同靶点;运用STRING数据库和Cytoscape 3.7.1软件构建共同靶点的蛋白-蛋白互作(PPI)网络并进行拓扑学分析;采用ClusterProfiler R功能包对共同靶点进行基因本体(GO)功能和KEGG通路富集分析;最后使用Cytoscape 3.7.1软件构建“活性成分-靶点-通路”网络并分析。结果筛选得到丹参-葛根药对候选活性成分30个,活性成分和心肌纤维化共同靶点41个。共同靶点PPI网络的平均点度值为19.7,平均介数为19.1,度值和介数均超过平均值的靶点共有14个。KEGG显著富集到73条通路,其中与心肌纤维化相关的通路有6条。“活性成分-靶点-通路”网络显示,丹参中的木犀草素、丹参酮IIA和葛根中的葛根素、β-谷甾醇等活性成分通过共同调控脂质与动脉粥样硬化、糖尿病并发症中的AGE-RAGE、血流剪切力与动脉粥样硬化、缺氧诱导因子-1(HIF-1)、肿瘤坏死因子(TNF)、白细胞介数-17(IL-17)等信号通路起到抗心肌纤维化的功效。结论揭示了丹参-葛根药对多成分、多靶点、多通路治疗心肌纤维化的作用特点,为进一步研究丹参-葛根药对治疗心肌纤维化的作用机制提供理论依据和新的思路。

基于Citespace的粉葛研究进展可视化分析

基于文献计量学方法,通过Citespace软件绘制知识图谱,以“粉葛”和“甘葛藤”为主题词,将采集到的1993~2022年377篇文献作为研究对象,对文献的发文量、作者、发文机构、期刊分布、等进行信息挖掘,并通过关键词聚类图谱、时间图谱、突现词分析等方法进行可视化分析。结果表明,粉葛相关文献年发文量总体呈现上升趋势,2020以来呈现快速增长态势。从事粉葛研究的主要机构包括江西中医药大学、广西农业科学院、广西大学等,合作单位和合作团队呈现明显的“地缘”和“学缘”特征。目前,粉葛相关文献40%发表在核心期刊上,其中,中医药类和食品加工类期刊收录的粉葛相关文献较多。研究内容从理论走向实践,研究热点从粉葛品质鉴定、粉葛栽培技术转向粉葛生药学、产业经济调研等;目前,淀粉、产量、多糖等成为突现关键词;关键词时区图结果显示葛根素、总黄酮、HPLC等关键词贯穿整个粉葛研究阶段,成为该领域的研究基石,反应了粉葛研究存在的一定问题。

Kudzu Forage Quality Evaluation as an Animal Feed Source

The nutritional value of kudzu (Pueraria lobata) was evaluated as a potential animal feed. Randomly selected samples from natural habitats with Jacksonville area (Alabama, USA) were utilized in the study. Kudzu leaves showed significantly higher dry matter and crude protein content than the stems;whereas, ADF fiber was significantly lower in the leaves. However, NDF and total digestible nutrients (TDN) were similar in the leaves and the stems. The leaf part of kudzu was especially rich in CP (24.46%) and met the requirements and recommendations for most ruminants. The TDN value for kudzu stem is 55.99%, which falls below but within the recommended range. Macronutrients concentration between the stem and leaf of kudzu, nitrogen, calcium, and magnesium showed a significantly higher in the leaf than the stem. However, phosphorous was similar in concentration in the stem and the leaf, and potassium was higher in the stem. In regard to the micronutrients, the only significant difference between the leaf and the stem was iron. However, iron, manganese, zinc, and copper were all above the recommended concentrations by the National Research Council. In general, kudzu can be considered a good source for animal feed, especially the leaf, which shows a higher value in most of the determined criteria in comparison to what was recommended by the National Research Council. This should provide a new outlook to kudzu, which has been considered a noxious weed and help in the utilization of kudzu in a positive way in the southern region of the US.

Interaction of the Bioherbicide Myrothecium verrucaria and Glyphosate for Kudzu Control

Kudzu is an exotic invasive weed in the southeastern U.S. that is difficult to control with current commercial herbicides. Some success for its control has been achieved using a bioherbicidal agent, Myrothecium verrucaria (MV). Spore and mycelial formulations of MV were tested alone and in combination with glyphosate for control of kudzu (Pueraria lobata) under greenhouse and field conditions in naturally-infested areas. In greenhouse and field experiments, kudzu control increased as the concentration of spores or mycelia increased. Glyphosate alone provided 10%, 35%, 50% and 60% control in field experiments at 0.25, 0.50, 0.75 and 1.0X rates, respectively and MV alone spores provided 15%, 50%, 65% and 85% control at 0.25, 0.50, 0.75 and 1.0X rates, respectively. However, when MV spores were combined with glyphosate, significantly higher control occurred than that caused by either component alone. Similar levels of control were observed for MV mycelial formulations applied alone or with glyphosate at equivalent concentrations of the fungus. The rate of disease progression was more rapid and severe at all fungal spore or mycelial formulations and herbicide rates when these propagules were applied in combination with glyphosate. In field tests, 24 h after application, only 20% of kudzu plants were severely damaged by MV alone (0.25X), whereas 80% were severely diseased when MV spores and glyphosate were mixed and applied at 0.25X rates each. A similar trend occurred with the MV mycelial formulation applied at these rates. Synergist interactions on kudzu control were observed, especially when lower levels of MV (spores or mycelia) and glyphosate were combined and applied to kudzu in the greenhouse or in the field. These results suggest that it may be possible to incorporate glyphosate to improve the bioherbicidal control potential and reduce herbicide and inoculum requirements of M. verrucaria spores or mycelium for controlling kudzu.

秦岭太白野葛根异黄酮成分的研究

目的：对秦岭太白野葛根（ＰｕｅｒａｒｉａＬｏｂａｔａ）的异黄酮成分进行分离鉴定。方法：以甲醇冷浸提取物，硅胶柱层析分离，理化常数和光谱数据确定异黄酮类化合物的结构。结果：分得６个异黄酮成分，分别鉴定为芒柄花素（Ⅰ）、大豆苷元（Ⅱ）、３＇－甲氧基葛根素（Ⅲ）、８－碳－芹菜糖（１→６）葡萄糖大豆苷（Ⅳ）、大豆苷（Ⅴ）和葛根素（Ⅵ）。结论：化合物（Ⅰ），（Ⅲ）和（Ⅳ）系首次从该植物中分得。

野生葛不同部位铜、锌、铁、钙、镁含量测定

用硝酸、高氯酸混合酸消解样品,采用原子吸收光谱法,测定了河南大别山区野生葛不同部位即葛根、葛藤、葛叶及花蕾中铜、锌、铁、钙、镁的含量,并对其结果进行了分析。结果显示:葛根、葛藤、葛叶及葛花中含有丰富的宏量元素钙、镁和有益的微量元素铜、锌、铁。铜的分布量依次为:叶>花蕾>葛藤>葛根;锌的分布量依次为:叶>花蕾>葛藤>葛根;铁的分布量依次为:葛根>葛藤>叶>花蕾;钙的分布量依次为:叶>葛根>花蕾>葛藤;镁的分布量依次为花蕾>葛根>叶>葛藤。测定结果为探讨中草药中宏量、微量元素与治疗心脑血管疾病功效的关系提供了有用的数据;测定结果还显示:葛叶中铜、锌、钙含量居其他部位含量之首,适宜大力开发葛叶产品。

野葛根异黄酮成分的超声萃取及抗氧化作用

采用超声法分别以95%乙醇、甲醇、70%乙醇和水为溶剂在不同时间水平从野葛根中萃取异黄酮活性成分,并用定性、定量的分析方法测定了野葛根提取物中异黄酮成分及其质量分数;同时,测定了在最佳条件下提取的葛根异黄酮对DDPH自由基的清除率。结果表明:以甲醇为溶剂超声萃取30分钟提取野葛根异黄酮成分效果最佳,提取率为20.6%,总黄酮质量分数为50.03%;采用超声萃取法从野葛根中提取异黄酮活性成分具有省时、节约能源、总黄酮提取率和产品纯度高的优点;葛根异黄酮具有较强的抗氧化作用,是一种天然的抗氧化剂。

葛根高效液相指纹图谱的研究

目的 采取高效液相色谱法建立野葛的指纹图谱 ,为野葛药材质量控制提供依据。方法 Agilent高效液相色谱仪 ,Zorbax XDB- C1 8(4.6 mm× 2 5 0 mm ,5μm)色谱柱 ,甲醇和水梯度洗脱 ,流速 0 .5 m L /min,检测波长 2 5 0nm。结果 精密度和重复性试验中各共有峰相对峰面积的 RSD均小于 5 % ,符合有关规定。

葛藤不同类型叶耐旱结构的比较解剖学研究

选取葛藤栅栏组织厚度/叶肉组织厚度、栅栏组织直径、海绵组织直径、气孔密度、表皮毛密度、维管束密度、主脉厚角组织层数等旱生指标,对葛藤的3个引种类型叶的旱生结构进行比较解剖研究,结果表明3个类型的相对抗旱顺序为:美国葛藤优势最强,粉葛次之,药葛最差.

葛根总黄酮提取工艺研究

本论文采用传统浸提法,以乙醇为溶剂,考察溶剂体积分数、提取时间、固液比、提取温度和葛根粉粒度对葛根总黄酮提取率的影响,并在单因素实验的基础上,通过正交试验,确定了葛根总黄酮的提取工艺。结果表明,提取温度、提取时间、固液比、乙醇体积分数以及粒度对葛根总黄酮的提取率均有影响,优化的葛根总黄酮的提取工艺为:提取温度为80℃,提取时间为120min,乙醇体积分数为70%,固液比为1:12,粒度为80目。

RP-HPLC法同时测定野葛的根、茎和叶中葛根素、大豆苷和大豆苷元的含量

目的:建立RP-HPLC法同时测定野葛的根、茎及叶中葛根素、大豆苷和大豆苷元的方法。方法:样品前处理采用70%乙醇超声提取,HPLC直接测定,并与药典方法进行对比。结果:对照品葛根素、大豆苷和大豆苷元分别在11.87～73.91,1.00～6.48和0.28～1.53μg/ml范围内呈良好线性关系,相关系数r分别为0.9998,0.9997和0.9994,最低检出限分别为0.43、0.24、0.18μg/ml。平均回收率分别为99.78%、102.61%和105.57%,RSD分别为1.6%、2.8%和3.4%(n=5)。精密度、稳定性和重现性良好。超声提取法测得的葛根中葛根素、大豆苷和大豆苷元含量分别为3.98%、0.36%和0.039%,高于药典提取法的提取率,更适于同时提取葛根素、大豆苷和大豆苷元。葛茎中葛根素、大豆苷和大豆苷元含量分别为0.004%、0.0032%和0.0007%,葛叶中葛根素、大豆苷和大豆苷元含量分别为0.0016%、0.005%和0.0012%。结论:该方法准确、快速,操作简单,分离效果好,可用于野葛的根、茎和叶中葛根素、大豆苷和大豆苷元含量的同时测定;江苏茅山地区野葛茎和叶中葛根素、大豆苷和大豆苷元含量大大低于葛根中的含量。

UHPLC法同时测定不同产地葛根中5种活性成分的含量

目的：建立同时测定不同产地葛根中3＇-羟基葛根素、葛根素、3＇-甲氧基葛根素、大豆苷和大豆苷元5个成分含量的UHPLC法。方法：采用Agela Venusil MP C18（100 mm×2.1 mm,3μm）色谱柱,以甲醇-0.1%甲酸梯度洗脱,检测波长250 nm。结果：3＇-羟基葛根素、葛根素、3＇-甲氧基葛根素、大豆苷和大豆苷元分别在12.41-248.24ng（r=0.9999）、58.82-1 176.47 ng（r=0.9997）、12.65-252.94 ng（r=0.9999）、12.14-242.82 ng（r=0.9998）、1.82-36.30 ng（r=0.9997）进样量范围内与峰面积呈良好的线性关系;平均加样回收率为99.03%-100.32%,RSD值为0.26%-1.37%。结论：本方法简便可靠、快速,可有效控制葛根的质量。

超声提取葛根总黄酮成分的研究

以乙醇为溶剂,研究了预浸时间、超声功率、超声作用时间以及占空比对葛根总黄酮提取率的影响,结果表明,预浸时间为10min,超声功率为300W,超声作用时间为20min,占空比为1时,所得到总黄酮的提取率最大。

葛根淀粉老化动力学的研究

利用宽角X 射线粉末衍射和差示扫描量热法研究了储存于不同温度下 ,随储存时间延长 ,葛根淀粉的老化度、老化速率等老化动力学问题。结果表明 ,完全糊化的葛根淀粉储存于 - 2 2℃ ,4℃和 2 8℃ ,均会发生老化作用而具有明显的X 射线衍射峰。所形成的老化淀粉衍射峰的衍射强度有较大差异。在 4℃储存时衍射峰的强度最大 ,2 8℃储存时衍射峰的强度最小 ,表明储存于 4℃时老化度最大 ,储存于 2 8℃时老化度最小 ,- 2 2℃的老化度居两者之间。在储存初期 (0～ 4d) ,老化度与储存时间基本呈正相关。根据Avrami方程计算的结果 ,储存于 4℃时的老化速率是 - 2 2℃储存时的 1.6倍 ,是 2 8℃储存时的 3.4倍。试验结果为生产具有较高储存稳定性和良好品质的葛根淀粉类食品提供理论依据。