葛根芩连颗粒的制备及质量检查

葛根芩连汤,出自于《伤寒杂病论》,原文为太阳病,桂枝证,医反下之,利遂不止,脉促者,表未解也,喘而汗出者,葛根黄芩黄连汤主之。现有市场销售品种,主要有葛根芩连颗粒、葛根芩连片、葛根芩连丸、葛根芩连口服等。该文主要讨论葛根芩连颗粒的制备及质量检查。     

葛根中葛根素和大豆苷提取方法的比较研究

目的:比较不同方法提取葛根中葛根素和大豆苷的效果。方法:分别用乙醇回流提取法、微波辅助提取法、超声波提取法及热水浸提法提取葛根中葛根素和大豆苷,用高效液相色谱法测定含量,计算提取量,并对最佳方法进行优化。结果:乙醇回流提取法、微波和超声法提取量较高,热水浸提法最低。超声提取法的最佳工艺为:以70%乙醇为溶剂,1:10料液比,提取时间45min。结论:该提取方法操作简便,提取效率高,提取时间短,节约溶剂,为工业化生产提供理论依据。     

葛根芩连汤加蒲公英灌肠治疗溃疡性结肠炎的临床观察

目的:观察中药葛根芩连汤加蒲公英灌肠治疗溃疡性结肠炎的疗效。方法:溃疡性结肠炎患者113例,随机分为治疗组(60例)和对照组(53例),前者用蒲公英葛根芩连汤灌肠,后者用柳氮黄吡啶治疗。结果:治疗组总有效率为90%,对照组总有效率75.5%,差异具有显著性(P<0.01)。结论:中药葛根芩连汤加蒲公英灌肠治疗溃疡性结肠炎疗效好,建议临床推广应用。     

酸水解法从葛根中提取分离葛根素和大豆苷元

本文报道了葛根黄酮的提取精制方法,以６０％乙醇为溶剂,６０℃温浸６ｈ的优化工艺条件下,采用逆流萃取法从葛根中提取葛根黄酮,其收得率为１９．２８％,含量５１．０５％。葛根黄酮提取物采用５％盐酸水解４ｈ,酸水解液用乙酸乙酯萃取放置析出葛根素,乙酸乙酯相经水洗、浓缩和重结晶可得大豆苷元。采用酸水解法可以将葛根黄酮中的葛根素及大豆苷元衍生物水解成葛根素和大豆苷元,有利于葛根素、大豆苷元的分离及产率的提高;该方法从葛根中提取分离葛根素和大豆苷元具有操作简便、产品纯度和产率高、成本低的特点。葛根素和大豆苷元产率分别为１．３６％和０．４５％,纯度为９８．３２％和９１．２５％。     

超声提取和超声水解法从野葛根中提取纯化葛根素和大豆苷元

采用超声法从野葛根中提取葛根异黄酮,再将提取物在盐酸水溶液中超声水解结合有机溶剂萃取法从野葛根中分离纯化葛根素和大豆苷元。葛根素收得率为1．2％,纯度为97．8％;大豆苷元收得率为0．5％,纯度为98．2％。超声法从野葛根中提取分离葛根异黄酮活性成分葛根素和大豆苷元具有省时、节能、提取率和产品纯度高的优点。     

高聚物型色谱柱在高效液相色谱法中一步分析制备葛根素及大豆苷元的应用

建立了使用以聚苯乙烯-二乙烯基苯(PS-DVB)为基质的高聚物型色谱柱一步分离制备葛根提取液中的葛根素和大豆苷元的方法。采用MKF-RP-HH色谱柱(300 mm×7.8 mm,8μm),葛根提取液经70%乙醇溶解稀释,以水(A)和甲醇(B)为流动相进行梯度洗脱,流速1.0 m L/min,检测波长250 nm,柱温30℃。结果表明:葛根提取液中的2种有效成分葛根素和大豆苷元均与杂质达到了较好的分离效果,且柱效大于2 100 N/m,优于常规的C-18硅胶色谱柱(柱效:1 000 N/m)。该高聚物型色谱柱不仅可用于上述分析,还可用作半制备柱制备得葛根素和大豆苷元纯品,葛根素在2~50μg/m L范围内线性关系良好,回归方程为y葛根=1.342 2x-1.018 4,线性相关系数为0.999;大豆苷元在2~20μg/m L范围内线性关系良好,回归方程为y大豆=2.275x+0.869 5,线性相关系数为0.999;葛根素和大豆苷元的纯化得率分别可达95.9%和78.5%。     

葛根芩连汤对肠道湿热证泄泻小鼠肠道微生物的影响

目的探讨葛根芩连汤对肠道湿热证泄泻小鼠肠道微生物的影响。方法造模:10只KM雄性小鼠随机分为正常组5只和模型组5只,采用"高糖高脂+高温高湿+白酒复合灌胃冰水"的方法制备肠道湿热证泄泻小鼠模型。治疗:15只KM雄性小鼠随机分为正常组5只和模型组10只,造模成功后将模型组小鼠分为自愈组和治疗组,治疗组小鼠运用葛根芩连汤进行治疗,分别在造模和治疗后无菌采集小鼠肠道内容物进行微生物培养。结果造模后,小鼠出现懒动、肛温升高、体重增长缓慢、黄褐色稀便、肛门污秽等湿热泄泻症状,肠道乳杆菌数高于正常组,而细菌总数、肠杆菌、双歧杆菌、B/E值降低,其中细菌总数以及双歧杆菌数与正常组相比差异有统计学意义(t=14.182,P0.001;t=2.797,P=0.049)。经葛根芩连汤治疗后,小鼠肠道乳杆菌和双歧杆菌数高于正常组,而细菌总数、肠杆菌数低于正常组,且乳杆菌数和肠杆菌数与正常组相比差异有统计学意义(t=-5.210,P=0.033;t=-11.160,P=0.008),B/E值与正常组差异无统计学意义(t=-1.427,P=0.227)。结论造模使小鼠肠道双歧杆菌数显著降低,B/E值下降;葛根芩连汤可促进肠道湿热证泄泻小鼠肠道乳杆菌和双歧杆菌的增生,提高B/E值。     

6种晋产野生葛根中葛根素、大豆苷和总黄酮含量比较

采用高效液相色谱法对山西省5地6处野生葛根中主要有效成分葛根素、大豆苷及总黄酮含量进行了分析,并对HPLC法的测定条件(流动相和洗脱程序)进行了优化。结果表明,6种晋产野生葛根中主要有效成分含量差异显著,葛根素的含量为1.46%~4.48%,大豆苷的含量为0.11%~0.62%,总黄酮的含量为11.19%~26.14%。葛根素、总黄酮含量与纬度高低呈正相关。晋南绛县么里镇小北山与夏县泗交镇太宽河自然保护区的野生葛根中葛根素含量低于《中国药典》规定的2.4%,而晋北灵丘县的野生葛根中葛根素含量则高达4.48%。     

葛根芩连汤在模拟胃肠液中对微生物生长的影响

目的探究葛根芩连汤在模拟胃肠液中对微生物生长的影响。方法在模拟胃肠液中分别加入消化液10%量的金黄色葡萄球菌、大肠埃希菌、沙门菌、产气杆菌、黑曲霉、米曲霉、青霉和汉逊德巴利酵母,然后加入生药浓度为1 g/mL的葛根芩连汤水煎液,37℃恒温培养。分别在0、1、2和4 h吸取1 mL培养液做稀释平板计数。结果体外模拟肠液中,葛根芩连汤对金黄色葡萄球菌、大肠埃希菌、沙门菌、产气杆菌、黑曲霉、米曲霉和青霉有较强的抑制作用,而对汉逊德巴利酵母的生长则为先促进后抑制,并且在2 h时对黑曲霉、米曲霉、青霉和汉逊德巴利酵母的抑制效果最显著;在模拟胃液中,葛根芩连汤对金黄色葡萄球菌、大肠埃希菌、沙门菌、黑曲霉、青霉和汉逊德巴利酵母均有一定的抑制作用,而对产气杆菌无明显影响。结论葛根芩连汤在模拟胃肠液中对金黄色葡萄球菌、大肠埃希菌、沙门菌、黑曲霉、米曲霉、青霉和汉逊德巴利酵母等均具有一定的抑制作用,在模拟肠液中对产气杆菌抑制作用明显,而在模拟胃液中对其抑制效果不明显。     

葛藤与葛根中异黄酮类成分的比较

分析比较了野葛〔Ｐｕｅｒａｒｉａｌｏｂａｔａ（Ｗｉｌｄ．）Ｏｈｗｉ〕的藤（葛藤）和根（葛根）的主要异黄酮类活性成分。从葛藤中首次分离得到３个化合物,经化学方法和光谱鉴定,证实为大豆甙元（Ａ）、大豆甙（Ｂ）和葛根素（Ｃ）。采用双波长薄层扫描法测定葛藤与葛根中上述３种异黄酮化合物的含量,葛藤中大豆甙元、大豆甙和葛根素的含量分别为０．１９５％,３．９３３％和２．４８１％;葛根中则分别为０．０５９％,０．７１４％和４．３１５％。研究结果为葛藤新药源的开发利用提供了科学依据     

汽爆葛根直接固态发酵乙醇联产葛根黄酮

针对葛根原料富含纤维和黄酮等特点, 选用低压汽爆处理代替糊化直接固态同步糖化发酵乙醇, 而后提取发酵剩余物的葛根黄酮。实验结果表明鲜葛根在0.8 MPa压力下维持3.5 min汽爆处理后, 直接加入糖化酶(65 u/g)、纤维素酶(1.5 u/g), 0.1%(NH4)2SO4、0.1%KH2PO4和活化后的酵母, 35~37oC下, 固态同步糖化发酵60 h, 100 g干葛根可生产的乙醇与葛根黄酮分别为27.47 g、4.43 g, 淀粉利用率达到95%。该方法实现了葛根分层多级转化清洁利用, 为非粮食类淀粉资源发酵乙醇提供了一条新途径。     

汽爆葛根直接固态发酵乙醇联产葛根黄酮

针对葛根原料富含纤维和黄酮等特点, 选用低压汽爆处理代替糊化直接固态同步糖化发酵乙醇, 而后提取发酵剩余物的葛根黄酮。实验结果表明鲜葛根在0.8 MPa压力下维持3.5 min汽爆处理后, 直接加入糖化酶(65 u/g)、纤维素酶(1.5 u/g), 0.1%(NH4)2SO4、0.1%KH2PO4和活化后的酵母, 35~37oC下, 固态同步糖化发酵60 h, 100 g干葛根可生产的乙醇与葛根黄酮分别为27.47 g、4.43 g, 淀粉利用率达到95%。该方法实现了葛根分层多级转化清洁利用, 为非粮食类淀粉资源发酵乙醇提供了一条新途径。     

纤维素酶解-超声偶联法提取葛根中总异黄酮的工艺优化

采用酶解-超声偶联新工艺对葛根中总异黄酮提取进行优化,以提高其收率.在单因素试验的基础上,利用Box-Behnken响应面试验设计,以蒸馏水作为提取溶剂,对温度、酶浓度、时间进行三因素三水平的试验设计优化.结果表明最佳提取工艺条件为:温度34℃、酶浓度0.63 mg/mL、时间62 min.在此条件下,葛根总异黄酮提取率为7.10％,与预测值的相对误差为0.34％.酶解-超声偶联工艺是一种新型、快速、有效的总黄酮提取方法.     

Seasonal variations in the isoflavonoids of radix Puerariae

The variations in isoflavonoid content in Radix Puerariae, the root of Pueraria lobata (Wild) Ohwi, have been examined in order to determine the optimum time to harvest the herb. Total isoflavonoid contents have been analysed by a UV spectrophotometric method whilst HPLC with photodiode array detection has been used to monitor the contents of seven major individual isoflavonoids, namely, 3'-hydroxypuerarin, puerarin, 3'-methoxypuerarin, daidzin, genistin, formononetin-7-glucoside and daidzein. Ninety-six samples of Radix Puerariae of different ages and harvested in different months were collected and analysed. The results clearly showed that 3-year-old roots harvested in January have the highest yields of isoflavonoid compounds. Moreover, the results obtained by both methods have a strong correlation. This suggested that the simple and fast UV spectrophotometric method could be used for monitoring the phytochemical quality of Radix Puerariae.     

Induction of growth hormone release by Pueraria thunbergiana BENTH.

Puerariae Radix (PR), Puerariae Flos (PF), and Puerariae Surculus (PS) as well as their constituents were tested for induction of rat growth hormone (rGH) release by both rat pituitary cell culture and in vivo experimentation in order to develop them to novel drugs. Through a calibration curve of the rGH released by addition of rat growth hormone-releasing hormone (rGHRH) to rat pituitary cells, the 70 % ethanol extracts of PR and PS increased rGH release by about 1.6 and 1.7 times as high, respectively, as the control group (264.6 +/- 13.6 pM). However, each puerarin type as a representative constituent of PR in Korea Pharmacopeia (KP) and tectorigenin and an important ingredient of PF were twice as effective as in the control group. The acid hydrolysate of Puerariae Surculus (HPS) increased rGH release concentration-dependently, and its EC (50) was approximately 10.4 microg/ml. The T (max) value for rGH after injection of 20 microg/kg of rGHRH was 10 - 30 min, while the C (max) value was increased by approximately 12-fold compared to the control group (198.2 +/- 25.0 pM) and the AUC (0 - 45) was increased to 10 times the level of the control group (10,840.9 +/- 845.5 min. pM). On the other hand, T (max) for the HPS was 60 min, while C (max) was increased approximately to 5.8 fold compared to control (244.1 +/- 36.4 pM). C (max) for puerarin was 1,028.6 +/- 502.7 pM, that is, approximately 5.2 times as high as the control level. However, tectorigenin (20 microg/kg) was of no statistical significance. Therefore, we suggest that the HPS and puerarin act either on GH secretagogue receptors or on GHRH receptor of somatotrophin as possible agonists or an inhibitor on somatostatin receptor to release rGH, respectively.     

葛根素对中枢神经系统神经细胞保护作用的研究进展

葛根素（puerarin）是从中药葛根中提取的主要有效成分之一,为一种异黄酮化合物。它具有抑制脑细胞凋亡,保护神经细胞的能力,还能够提高神经元抵抗损伤的能力,可用于治疗脑损伤以及神经退行性疾病。本文就葛根素对中枢神经系统神经细胞的保护作用及机制方面的研究进展做一综述。     

Effects of radix polygoni multiflori components on tyrosinase activity and melanogenesis

Radix Polygoni multiflori is a herb used effectively to prevent graying and treat skin depigmentation diseases in traditional Chinese medicine but its active ingredients have not been discovered yet. In this investigation, we tested six compounds isolated from Radix Polygoni multiflori, to discover the active component on melanogenesis. Three experiments were performed in the present investigation: mushroom tyrosinase activity, melanin content B16 cell proliferation assay. Among all the six components tested, THSG showed the most potent effects on tyrosinase activation and melanogenesis; it was shown to be a potent tyrosinase activator and a melanogenesis stimulator in this study. On the other hand, we found that gallic acid significantly inhibited tyrosinase and, in addition, anthraquinones were cytotoxic to melanoma cells. They were both harmful to melanogenesis. Therefore, we propose that THSG acts as the active ingredient of Radix Polygoni multiflori on melanogenesis.     

Main antimicrobial components of Tinospora capillipes, and their mode of action against Staphylococcus aureus

In this investigation, the antibacterial modes of action of Radix Tinosporae, its major single components, and nine antibiotics with different targets or modes-of-action on Staphylococcus aureus were studied. Metabolic profiles of cultures treated with different medicines were acquired by HPLC/ESI-MS. After HPLC-MS data pretreatment, those profiles acquired were reduced into several MS vectors. Then statistical processing by principal components analysis was carried out upon those vectors, two conclusions could be drawn: (1) the antibacterial mode of action of Radix Tinosporae is similar to that of rifampicin and norfloxacin, which act on nucleic acid; (2) its active components playing main antimicrobial roles on Staphylococcus aureus might be alkaloids, such as palmatine and jatrorrhizine.     

Effects of radix polygoni multiflori components on tyrosinase activity and melanogenesis

Radix Polygoni multiflori is a herb used effectively to prevent graying and treat skin depigmentation diseases in traditional Chinese medicine but its active ingredients have not been discovered yet. In this investigation, we tested six compounds isolated from Radix Polygoni multiflori, to discover the active component on melanogenesis. Three experiments were performed in the present investigation: mushroom tyrosinase activity, melanin content B16 cell proliferation assay. Among all the six components tested, THSG showed the most potent effects on tyrosinase activation and melanogenesis; it was shown to be a potent tyrosinase activator and a melanogenesis stimulator in this study. On the other hand, we found that gallic acid significantly inhibited tyrosinase and, in addition, anthraquinones were cytotoxic to melanoma cells. They were both harmful to melanogenesis. Therefore, we propose that THSG acts as the active ingredient of Radix Polygoni multiflori on melanogenesis.     

不同干燥方法对麦冬多元活性成分的影响

探究不同干燥方法对麦冬多元活性成分的影响。采用超快速液相色谱-三重四极杆/线性离子阱质谱(UFLC-QTRAP-MS/MS)同时测定不同干燥方法下麦冬中甾体皂苷、高异黄酮、氨基酸及核苷类共29种活性成分的含量;根据29种目标成分的含量,用聚类分析(HCA)及主成分分析(PCA)对不同干燥方法下麦冬进行综合评价。结果表明,29种目标成分在一定浓度范围内线性关系良好,相关系数均大于0. 999 0;精密度、重复性、稳定性良好,RSD均小于5%;平均加样回收率为97. 02%～102. 78%,RSD均小于5%。PCA结果显示,以晒干、无硫烘干的麦冬样品综合质量较优。所建立的方法准确、可靠,可用于麦冬药材内在质量的综合评价,该研究可为揭示麦冬传统干燥方法科学内涵提供依据,同时为麦冬产地加工时干燥方法的优选提供基础资料。