赤芍和白芍新鲜花瓣正己烷提取成分GC-MS分析

目的:赤芍和白芍新鲜花瓣正己烷提取成分比较分析。方法:正己烷浸出法提取新鲜花瓣挥发性物质,GC-MS分析、鉴定其化学成分。结果:从赤芍新鲜花瓣挥发性物质中鉴定出33种化学成分;白芍新鲜花瓣挥发性物质中鉴定出35种化学成分。二者中的主要成分为棕榈酸,二十三烷,二十五烷,二十七烷,二十九烷等。赤芍花瓣含有更多小分子芳香类成分,如苯乙醇、法尼醇类等;白芍缺少这些成分,其芳香类成分有β-沉香醇、反式-橙花椒醇等。结论:赤芍(野生芍药)与白芍(栽培芍药)花瓣芳香气成分有差异。     

近红外光谱法快速测定白芍中芍药苷含量

本研究旨在应用近红外光谱法建立一种白芍药材中芍药苷含量的快速测定方法。利用HPLC测定样品中芍药苷含量,并以其作为参考值,运用偏最小二乘法(PLS)建立芍药苷含量与近红外光谱之间的多元校正模型,对未知样品进行含量预测。结果表明,所建芍药苷定量分析模型的相关系数(R2)、内部交叉验证均方差(RMSECV)、校正均方差(RMSEC)分别为0.99395、0.33068、0.0563;经内部验证,模型的预测均方差(RMSEP)和平均回收率分别为0.0756和100.07%。该方法操作简便,无污染,结果准确可靠,可用于白芍中芍药苷含量的快速测定。     

白芍总苷在正常大鼠体内的组织分布研究

探讨白芍总苷在正常大鼠体内的组织分布特点,为预测其药理作用及不良反应提供依据.正常大鼠按2.82 g/kg灌胃给予TGP药液后1、3、6h取心、肝、脾、肺、肾、胃、小肠、大肠等组织,各组织匀浆后,将匀浆液制成冻干粉,HPLC法测定冻干粉中芍药苷和芍药内酯苷浓度,计算各组织中两者浓度.结果显示1h各组织中均能测到芍药苷和芍药内酯苷,3h除胃和小肠外,其他各组织中两者浓度均达到最大值,小肠、胃、大肠及肾、脾、肝中浓度较高,6h小肠、大肠、胃中浓度较高,其他各组织中浓度较低.说明灌胃TGP后组织分布迅速且广泛,胃、小肠、大肠及肾、脾、肝是主要分布器官,容易在胃肠蓄积,其他组织中蓄积较少,为进一步研究白芍总苷的药理作用及作用机理提供了指导,同时为白芍归经理论提供了一定的现代科学依据.     

赤芍中芍药苷提取工艺的优化

本文提出一条从赤芍中提取芍药苷的工艺,包括醇水溶液回流提取、液液分配、硅胶柱层析等工序,并对各工序的操作条件进行了优化。采用优化后的提取工艺,可得到纯度大于97.0%的芍药苷产品,总回收率超过87.0%。本工艺具有产品纯度高、操作简单、成本低、回收率高、易于工业化等优点。     

HPLC特征图谱结合含量测定的白芍及其炮制品的质量对比研究

建立白芍、炒白芍、酒白芍、硫熏白芍HPLC特征图谱,并结合多成分含量测定,为白芍、炒白芍、酒白芍和硫熏白芍的质量控制提供参考。采用Intersustain C₁₈(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.1%醋酸水溶液,流速为每分钟1 mL,梯度洗脱,检测波长为230 nm,柱温为30℃,进样量为10μL。14批白芍、炒白芍、酒白芍和硫熏白芍的特征图谱,标定了6个共有峰,并均被指认,分别为没食子酸、儿茶素、芍药内酯苷、芍药苷、1,2,3,4,6-五没食子酰葡萄糖和苯甲酰芍药苷,而硫熏白芍标定7个共有峰,峰7为白芍硫熏后产生;且各色谱谱峰有较好的分离,但不同炮制品特征图谱存在一定差异;含量测定结果显示,白芍炒制、酒制及硫熏后,6种成分均有不同程度的变化;借助中药色谱指纹图谱相似度评价系统和SIMCA-P13.0软件对14批白芍、炒白芍、酒白芍和硫熏白芍进行相似度和正交偏最小二乘判别(OPLS-DA)分析,所建立的白芍和炮制品及硫熏品的质量评价方法稳定性、重复性好,可用于白芍、炒白芍、酒白芍和硫熏白芍的质量控制和评价。     

基于HPLC指纹图谱的亳白芍不同炮制品标准汤剂7个成分含量测定

建立亳白芍不同炮制品标准汤剂HPLC指纹图谱,其中生白芍、炒白芍和酒白芍标准汤剂分别标定16、14和13个共有峰,同时测定亳白芍不同炮制品标准汤剂中7种化学成分(没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酸、1,2,3,4,6-五没食子酰葡萄糖和苯甲酰芍药苷)含量,经炒制后标准汤剂中芍药内酯苷和苯甲酸含量升高,氧化芍药苷、芍药苷含量降低,酒白芍标准汤剂中芍药苷含量升高,苯甲酸含量降低,并对含量测定结果进行聚类分析,同一批亳白芍及其炮制品可聚为一类,但聚类距离缩小后生品和炮制品各聚为一类。建立的HPLC指纹图谱和含量测定方法具有良好的重现性,且简便快速,二者结合可直观反映出亳白芍炒制、酒制后的变化差异。     

高效液相色谱法测定妇血荣胶囊中芍药苷的含量

目的:建立妇血荣胶囊中芍药苷含量的HPLC检测方法.方法:采用甲醇提取样品;色谱条件:Alltech ODS柱(5μm,250mm×4.6 mm);流动相为乙腈-水(16:84,v/v);检测波长为230 nm.结果:线性范围为24.3～218.7 μg.mL-1(r=0.9999),平均回收率为101.21%,RSD为0.80%.结论:高效液相色谱法简单易行,准确,灵敏度高,适用于妇血荣胶囊中芍药苷的含量测定.     

UPLC-MS同时测定赤芍中五个指标成分的含量

建立超高效液相色谱-串联质谱法同时测定赤芍药材中的没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷含量的分析方法。采用Acquity UPLC BEH C18柱(2.1 mm×50 mm,1.7μm);流动相为0.1%甲酸乙腈溶液(A)-0.1%甲酸水溶液(B),梯度洗脱,电喷雾离子源(ESI),选择性离子监测(SIR)模式进行正负离子同步监测。没食子酸、氧化芍药苷、芍药内酯苷、芍药苷和苯甲酰芍药苷在0.134~31.250、0.014~2.784、0.471~30.750、0.293~75.040和0.158~40.360μg/m L浓度范围线性关系良好,平均加样回收率为91.36%~112.83%,RSD为1.1%~8.9%。该方法准确、高效、重现性好、专属性高,可用于赤芍药材的质量控制。     

UPLC-MS同时测定赤芍中五个指标成分的含量北大核心CSCD

建立超高效液相色谱-串联质谱法同时测定赤芍药材中的没食子酸、氧化芍药苷、芍药内酯苷、芍药苷、苯甲酰芍药苷含量的分析方法。采用Acquity UPLC BEH C18柱（2.1 mm×50 mm,1.7μm）;流动相为0.1%甲酸乙腈溶液（A）-0.1%甲酸水溶液（B）,梯度洗脱,电喷雾离子源（ESI）,选择性离子监测（SIR）模式进行正负离子同步监测。没食子酸、氧化芍药苷、芍药内酯苷、芍药苷和苯甲酰芍药苷在0.134~31.250、0.014~2.784、0.471~30.750、0.293~75.040和0.158~40.360μg/m L浓度范围线性关系良好,平均加样回收率为91.36%~112.83%,RSD为1.1%~8.9%。该方法准确、高效、重现性好、专属性高,可用于赤芍药材的质量控制。     

传统中药白芍原植物分类鉴定及根形态解剖研究

对我国主产地的白芍(Radix Paeonia alba)原植物10个居群进行调查及标本采集,植物鉴定结果为：川白芍原植物粉红花居群为原变种芍药Paeonia lactiflora Pall,白花居群为芍药变种毛果芍药P.lactiflora var.trichocarpa(Bunge)Stern;毫白芍原植物线条居群和蒲棒居群均为芍药;杭白芍原植物红花、白花、粉红花居群均为毛果芍药;陕西韩城、江苏东海、山东荷泽白芍均为芍药,各居群在花色及形态上有明显而稳定的变异。根横切面解剖结构显示,按木质部的排列方式可将白芍原植物10个居群分为两大类：第一类为有呈两个不相连的扇形中央导管群,并且有狭长、具分枝的从形成层到根中央部分连续排列的木质部,基本上是原植物芍药的植物特征,其中毫白芍线条居群兼有毛果芍药和芍药的特征;第二类为具有不明显的根中央扇形导管束,导管束呈环状围绕中央排列,并且有粗短不分枝的靠近形成层处成群的导管,与原植物毛果芍药的特征基本一致,其中杭白芍红花居群与川白芍白花居群根中央导管群呈现较明显的2个分离扇形排列,类似芍药。白芍原植物的种分类定位与花色无密切关联,但由于性状的稳定,可以考虑作为变种或变型定位。     

高速逆流色谱法从白芍中分离纯化五没食子酰基葡萄糖

本文建立高速逆流色谱(HSCCC)方法,从白芍粗提物中分离纯化五没食子酰基葡萄糖.分别采用正己烷-乙酸乙酯-甲醇-水体积比0.5∶5∶1∶5及0.5∶5∶0.5∶5混合溶剂作为两相溶剂体系,上相为固定相,下相为流动相,转速为800 rpm,流速为2.0 mL/min,用HPLC检测及ESI-MS进行验证.经过两次HSCCC分离纯化,得到五没食子酰基葡萄糖纯度为95.7％.     

Immunoregulation and antioxidant activities of a novel acidic polysaccharide from Radix Paeoniae Alba

Radix Paeoniae Alba is widely used in Chinese traditional medicine to treat various diseases such as gastrointestinal disorders, immunomodulatory, cancer, and other diseases. In this paper, a novel acidic polysaccharide RPAPS purified from Radix Paeoniae Alba was evaluated for its structural features and potential of immunomodulatory and antioxidant activities. RPAPS (molecular weight: 1.0× 105 Da) was mainly composed of α-(1 → 4)-Glcp, α-Arap, α-Galp, α-Rhap, β-D-Glcp, α-(1 → 6)-linked Glcp and GalA. Immunological tests indicated that RPAPS could improve RAW264.7 phagocytic activity and LPS-induced splenocyte proliferation. For antioxidant activities, RPAPS showed reducing power and DPPH scavenging activity in dose dependent. Moreover, RPAPS could significantly protect the PC12 cells from H2O2 damage. These data implied polysaccharides RPAPS had the potential to be novel natural antioxidative and immunopotentiating agents for using in functional foods or medicine.     

基于ISSR与RAPD标记分析内蒙古赤芍的遗传多样性

采用ISSR和RAPD分子标记技术对28个赤芍种群进行遗传变异和亲缘关系分析,为准确地评价赤芍种质的遗传特征、资源保护及新品种选育提供理论依据。结果显示:(1)利用分别筛选的14条ISSR和RAPD引物扩增出257条和215条条带,其中多态性条带分别为251条和209条,多态性条带百分率分别为97.8%和97.2%;同时证实野生赤芍种群的遗传多样性高于栽培种群。(2)根据Shannon’s信息指数(I)和Nei’s基因多样性指数(H_e)值,发现内蒙古多伦种群(DL)的遗传多样性水平最高,建议在此地建立野生赤芍资源保护区。(3)根据遗传分化系数(G_(st)),发现野生赤芍种群的遗传分化主要发生在种群内,可能由遗传漂变引起;而栽培赤芍种群的遗传分化主要在种群间,说明栽培赤芍种群间的基因交流较少。(4)两种分子标记的聚类分析结果均将28个赤芍种群聚为5大类,遗传距离变化范围分别为0.115 1～0.343 8和0.095 5～0.286 2。研究表明,互相印证的ISSR和RAPD方法可以在DNA水平上更准确有效地分析赤芍种质资源的遗传结构和遗传多样性。     

Determination of paeoniflorin in rat hippocampus by high-performance liquid chromatography after intravenous administration of Paeoniae Radix extract

A rapid and simple high-performance liquid chromatographic (HPLC) assay for the determination of paeoniflorin in rat hippocampus was developed in this study. The chromatographic analysis was carried out using reversed-phase isocratic elution with a Zorbax SB-C(18) column, a mobile phase of methanol-water (32:68, v/v), and detection by ultraviolet (UV) absorption at 233 nm. The lower limits of quantitation (LLQ) were 1 microg/ml for paeoniflorin. The calibration curve for paeoniflorin was linear (r = 0.9999) over the concentration range of 1-50 microg/ml. The coefficients of variation of intra- and inter-day assays were 7.00, 0.58, 1.46% and 5.48, 1.79, 1.70% at concentrations of 1, 10, 50 microg/ml, respectively. The recoveries of paeoniflorin from rat hippocampus were 98.28 +/- 2.14, 98.96 +/- 1.48, and 95.34 +/- 0.92 at concentrations of 1, 10 and 50 microg/ml, respectively. Stability studies showed that paeoniflorin was stable at temperatures of 2-8 degrees C in methanol for at least 20 days. The method was applied to determine the time course of paeoniflorin in rat hippocampus, following the administration of a 60 mg/kg i.v. dose of paeoniflorin in Paeoniae Radix extract to a male Wistar rat.     

Protection of H9c2 rat cardiomyoblasts against oxidative insults by total paeony glucosides from Radix Paeoniae Rubrae

Total paeony glucosides (TPG) extracted from the roots of Radix Paeoniae Rubrae, have been approved for the therapy of rheumatoid arthritis by the State Food and Drug Administration. We previously demonstrated the myocardial protective effects of TPG in both isoprenaline-induced myocardial ischemia rat and acute myocardial infarction rat. However, the underlying mechanism of TPG effect in cardiomyocytes remains to be investigated. The aims of this study were to elucidate the effect of TPG on the activities of antioxidant defense targets and the bioenergetic system in rat cardiomyocytes. The changes of viability, antioxidant defense system activities, protein contents, and mitochondrial functions in tert-butyl hydroperoxide challenged H9c2 rat cardiomyoblasts were evaluated. The results suggest that TPG ameliorated cardiomyoblast dysfunction by preserving antioxidant defense and bioenergetic system.     

Paeoniae Radix,a Chinese herbal extract,inhibit hepatoma cells growth by inducing apoptosis in a p53 independent pathway

Paeoniae Radix (PR) is the root of traditional Chinese Herb named Paeonia lactiflora Pallas, which is commonly used to treat liver diseases in China for centuries. Several earlier studies have indicated that PR has anticancer growth activities, however the mechanism underlying these activities was unclear and remained to be elucidated. In this study, we evaluated the molecular mechanism of the effect of PR on human hepatoma cell lines, HepG2 and Hep3B. Our results showed that the water-extract of Paeoniae Radix (PRE) had inhibitory effect on the growth of both HepG2 and Hep3B cell lines. The induction of internucleosomal DNA fragmentation and chromatin condensation appearance, and accumulation of sub-G1 phase of cell cycle profile in PRE treated hepatoma cells evidenced that the cytotoxicity of PRE to the hepatoma cells is through activation of the cell death program, apoptosis. The activation of apoptosis by PRE is independent of the p53 pathway as Hep3B cell is p53-deficient. In addition, the differential gene expression of PRE treated HepG2 was examined by cDNA microarray technology and RT-PCR analysis. We found that the gene expression of BNIP3 was up-regulated while ZK1, RAD23B, and HSPD1 were down-regulated during early apoptosis of the hepatoma cell mediated by PRE. The elucidation of the drug targets of PR on inhibition of tumor cells growth should enable further development of PR for liver cancer therapy.     

当归及其混淆品独活、欧当归的紫外鉴别

为寻找当归及其混淆品独活、欧当归的紫外吸收光谱鉴别特征，采用紫外谱线组法对三者在不同极性溶剂中的紫外吸收光谱图进行比较。结果表明，当归和独活在四种极性溶剂中均存在明显差异，而当归和欧当归仅在无水乙醇和蒸馏水溶剂中鉴别差异显著。紫外谱线组法可以鉴别当归及其混淆品独活、欧当归。