固定化脂质体色谱与动物小肠吸收模型的相关性研究方法及其应用

固定化脂质体色谱(ILC)是一种新颖的筛选经被动吸收药物的技术,建立了ILC与常用的动物小肠吸收模型的相关性研究方法,考察了当归补血汤在ILC上有保留的成分的保留值与其在大鼠在体和离体小肠吸收模型中被吸收值的相关性。结果显示,当归补血汤在ILC上有保留成分的保留值与其通过大鼠在体小肠灌流和离体小肠实验测得的吸收值具有很好的相关性,其相关系数分别为0．9356和0．9350;当归补血汤在ILC上有保留的成分主要以被动扩散的方式被机体吸收。     

高效液相色谱与质谱联用分析测定川芎中有效成分阿魏酸与藁本内酯

采用HPLC-MS系统地研究了中药川芎有效成分阿魏酸(ferykuc acid)和藁本内酯(liguatilide)的定量分析方法。利用制备色谱制备了藁本内酯对照品,并对其进行了紫外光谱、质谱、红外光谱等结构鉴定。分别考察了水、甲醇、乙醇、95％乙醇4种溶剂以及提取时间对川芎中阿魏酸和藁本内酯提取量的影响。结果表明：水是阿魏酸的最佳提取溶剂,提取时间45min为宜;乙醇是适合提取藁本内酯的溶剂,提取时间75min为宜。以外标法对市售川芎中的阿魏酸与藁本内酯进行了定量分析,二者含量分别是0．15％(m／m)和0．82％(m/m)。     

分光光度法测定当归补血汤及其组分单味药提取液的抗氧化能力

对复方当归补血汤及其组成单味药即当归和黄芪的水提取液,以及两者的单味药的水提取液及乙醇提取液的抗氧化能力作了比较研究。将由Fenton反应产生的羟自由基在pH4.6乙酸盐缓冲溶液与水杨酸反应,生成紫红色的2,3-二羟基苯甲酸,其吸收峰在510nm波长处。当有任一抗氧化组分存在于此反应体系时,由于其与反应体系中的羟自由基的优先反应,使此体系在510nm处的吸光度下降。据此,对上述药物提取液的抗氧化能力进行了试验。所得结果表明:上述药物的提取液均呈良好的抗氧化能力(即清除羟自由基的能力),而复方当归补血汤中主成分药物的提取液(无论是水或乙醇的提取液)的抗氧化能力均高于单味药物的提取液。比较水提取液与乙醇提取液的抗氧化能力的结果表明,前者的效果更好。     

联用色谱及多组分光谱相关色谱用于当归补血汤与单味药成分的比较研究

摘要利用HPLC-DAD提供的二维化学数据信息, 通过多组分光谱相关色谱方法及化学计量学分辨技术, 对相同试验条件下所得的当归补血汤及其单味药的色谱组分进行了比较和归属分析. 结果表明， 在当归补血汤色谱响应值较高的21个色谱峰中, 8个色谱峰的组分来自当归, 6个色谱峰的组分来自黄芪, 7个色谱峰的组分同时来源于当归和黄芪. 其中一些色谱峰簇中的其它组分发生了消长变化， 既有新物质的产生, 又有原有化学成分的消失， 说明中药复方配伍的药效物质基础不仅是各单味药化学成分的总和, 还有单味药在复方配伍过程中各种成分的相互作用. 本方法为中药复方与单味药化学组分的比较及归属分析, 尤其对重叠色谱峰簇的组分比较分析提供了方便、 快捷及可行的研究方法.     

生物色谱用于当归中活性成分筛选及分离研究

应用分子生物色谱技术研究了筛选和分离当归中的活性成分。以甲基丙烯酸缩水甘油酯为功能单体,通过原位聚合得到了整体柱基质,用二乙胺活化后,以吸附蛋白的方法得到了以牛血清白蛋白(BSA)为固定相的分子生物色谱柱。当归样品经甲醇提取,0.45μm膜过滤后,以BSA分子生物色谱柱为分离柱,以含2%(体积分数)甲醇的Tris-盐酸缓冲溶液为流动相进行分离,用二极管阵列检测器检测。结果表明:活性组分在BSA上的保留机理是疏水作用、静电作用和特定的与空间结构相关的多种因素的共同作用结果。将BSA柱上的保留组分进行了气相色谱-质谱分析,鉴定了8种成分,包括当归中的主要可挥发性成分(藁本内酯)。     

新型大孔聚合物小球NKG-1载体在甲醇和乙醇分离测定中的应用

甲醇中微量乙醇及乙醇中微量甲醇的分离与分析一直是人们颇为关注的问题。由于高含量的峰易将低含量的峰掩盖,使二者难于分离或分离度很小。文献报道,选用D-山梨醇为固定液,6201为载体需填装5—8m长色谱柱方可使乙醇和甲醇较好分离;选用聚乙二醇-20M为固定液,硅烷化白色405为载体,分离白酒中各组分,甲醇和乙醇色谱峰分离度很小;Mangani等人应用色谱法直接测定果酒中甲醇含量,检测限为≈0.05％。 本文采用南开大学高分子化学研究所研制的新型大孔聚合物小球NKG-1为载体,涂渍10％聚乙二醇-20M,仅用2m长不锈钢柱,可将甲醇和乙醇较好分离,用于定量分析其准确度、精密度均较好,检测限可达0.005％。     

茶叶样品中绿原酸含量的高效液相色谱法测定

建立了茶叶绿原酸含量测定的择相高效液相色谱法,采用PecosphereC18柱,以甲醇－水（体积比18：20,水中含φ＝1／61的甲酸）淋洗,用紫外检测器于326nm处测定,各组分的色谱峰达到基线分离。绿原酸在0．02－4．02μg范围内线性关系良好,线性相关系数 r＝0．9996。在安徽黄山产贡菊和安徽产金银花中加标的回收率分别为86．8％、102．0％,该法已用于十多种茶趺实际样品的测定。     

高效液相色谱法研究当归指纹图谱

目的:研究当归药材的指纹图谱.方法:高效液相色谱法,Hypersil ODS柱(4.6 mm×250 mm,5μm),Kromasil ODS保护柱(4.6 mm×10 mm,5μm).甲醇-1%醋酸梯度流动相,流速1.0 mL/min,柱温25℃.以当归对照药材为对照品,色谱峰光谱采集范围:190~400 nm;以阿魏酸为参照物并测定了阿魏酸的含量,检测波长:323nm,流动相:甲醇-1%醋酸(40∶60,V/V),流速0.7 mL/min.结果:找出了22个共有峰,其中5号峰为阿魏酸,11个样品与当归对照药材之间的相似度均在90%以上,平均相似度为96.77%.结论:样品处理方法简单,研究所得的当归指纹图谱稳定性、重复性好,可以作为当归极性部分的特征性指纹图谱.     

中药配伍的统一分析方法研究 Ⅱ.拟合定量

 配伍过程中物质组分的浓度变化需要进行定量 ,但复方中大量的重叠色谱峰影响定量的准确性。而在色谱理论基础上发展起来的拟合定量技术 ,可以对重叠色谱峰进行拟合定量。在统一分离和复方中单味药的特征物质组分得到识别的基础上 ,以黄芪、当归用量变化时 ,复方“当归补血汤”中黄芪、当归的特征物质组分的拟合定量为实例 ,介绍了中药配伍过程中定量的手段和研究方法     

HPLC法测定鸡精中谷氨酸钠的含量

以邻苯二甲醛与谷氨酸钠中的氨基进行柱前在线衍生化反应,采用C18色谱柱分离、荧光检测器（激发340nm,发射450nm）进行测定,建立了柱前衍生反相高效液相色谱测定鸡精中谷氨酸钠含量的方法。该方法相对标准偏差为0．69％,加标回收率为99．1％～101％,在0．10~50．0mg／L范围内,谷氨酸钠的峰面积和浓度之间的相关系数为0．9999,保留时间和峰面积的相对标准偏差分别为1．22％和0．71％,鸡精中谷氨酸钠定量下限为0．2μg／g。     

以人血清白蛋白为固定相的分子生物色谱分析几种中药活性成分的研究

提出用人血清白蛋白分子生物色谱分析中药活性成分的方法,分别比较了当归等4种单味中药的分离结果,考察了提取当归的3种溶剂及提取时间对活性物质提取量的影响,同时建立了当归中阿魏酸与藁本内酯的定量分析方法.     

Screening and analysis of biologically active compounds inAngelica sinensis by molecular biochromatography

Summary A novel strategy for the screening and analysis of biologically active compounds in traditional Chinese medicine by molecular biochromatography is proposed. Molecular biochromatography with human serum albumin (HSA) immobilized on silica as stationary phase was used to screen and analyse the bioactive compounds in the typical Chinese medicine ofAngelica sinensis (Oliv.) Diels. Ten peaks showed retention on this column, which is based on their affinity for HSA. Ferulic acid and liguistilide were identified as the principal active components, which agrees very well with the results in the literature. A quality control method was also developed based on the simultaneous determination the concentrations of ferulic acid and liguistilide in solutions ofAngelica sinensis (Oliv.) Diels extracted with water and methanol. It was observed that the concentrations of ferulic acid and liguistilide in solution extracted with methanol were 2 and 53 times higher, respectively, than those with water. It was shown that molecular biochromatography is an effective way of analysing and screening biologically active compounds in traditional Chinese medicine.     

Assay of free ferulic acid and total ferulic acid for quality assessment of Angelica sinensis

Activity of Chinese Danggui (DG), the processed root of Angelica sinensis (Oliv.) Diels, is linked to the ferulic acid content but the stability of ferulic acid during extraction for medicinal use is not known. The stabilities of ferulic acid and coniferyl ferulate were evaluated in the extracts of DG using a variety of extraction solvents. These included various combinations and proportions of methanol, water, formic acid, 1 M aqueous hydrochloric acid and 2% sodium hydrogen carbonate (NaHCO3) in water. Coniferyl ferulate was found liable to hydrolyze into ferulic acid in neutral, strongly acidic and basic solvents, where heat and water could facilitate this hydrolysis. However, the hydrolysis was relatively resisted in weakly organic acid. Based on the stability evaluation, two new terms, namely: free ferulic acid and total ferulic acid, were suggested and defined. Free ferulic acid refers to the natural content of ferulic acid in herbs. Total ferulic acid means the sum of free ferulic acid plus the amount of related hydrolyzed components. Meanwhile, the high-performance liquid chromatographic (HPLC) method was developed to assay free ferulic acid and total ferulic acid in DG using methanol-formic acid (95:5) and methanol-2% NaHCO3 in water (95:5) as extraction solvents, respectively. Ten DG samples were investigated on their contents of free and total ferulic acid. The results indicated that the amount variety of free ferulic acid was larger than that of their counterparts, and the ratio of total ferulic acid to free ferulic acid was 4.07 +/- 2.73 (mean +/- SD, n = 10). The chemical assay of DG using total ferulic acid content would be a better choice to assess the herbal quality and was recommended.     

Extraction of ferulic acid from Angelica sinensis with supercritical CO2

Extraction of pharmacologically active ingredient of ferulic acid from the root of Angelica sinensis with supercritical CO2 was investigated. The experimental results show that the extract yields were 0.87-4.06% at temperatures from 45 to 65 degrees Celsius and pressures from 30 to 50 MPa, and the maximum content of ferulic aicd in the extracts was about 0.35-0.37%, which is lower than that of 0.61-0.85% by conventional percolation methods. Ethanol was used as co-solvent in different ratios to raw materials in order to increase the content of ferulic acid in the extracts. The experimental results show that both the extract yields and the content of ferulic acid in the extracts increase greatly compared with pure CO2 extraction. When the ratio of ethanol to the raw material was 1.6, the content of ferulic acid in the extracts was 0.91-1.27%, indicating that supercritical fluid extraction (SFE) with CO2 in the presence of suitable co-solvent is superior to percolation in extracting polar ferulic acid from Angelica sinensis.     

The difference between blood‐associated and water‐associated herbs of Danggui‐Shaoyao San in theory of TCM,based on serum pharmacochemistry

Danggui‐Shaoyao San (DSS) is a famous Chinese formula for activating blood circulation and promoting urination. This study was to investigate the difference of material basis between a blood‐associated herbs group and a water‐associated herbs group. According to the theory of traditional Chinese medicine, the formula can be divided into a blood‐associated herbs group (Angelica sinensis, Paeonia lactiflora and Ligusticum chuanxiong) and a water‐associated herbs group (Atractylodes macrocephala, Alisma orientale and Poria cocos). The HPLC fingerprint of the formula was established for quality control. Serum samples from rats, orally administrated DSS, and the decomposed recipes of DSS, were analyzed by HPLC‐DAD and the transitional blood components of DSS were identified. Twenty‐one common peaks were identified in the fingerprint of DSS. Contents of paeoniflorin, albiflorin, ferulic acid and alisol B 23‐acetate in co‐decoction were significantly higher than those in individual decoction. Eleven peaks belonged to the blood‐associated herbs group (four metabolites and seven prototype components; paeoniflorin and ferulic acid appeared in prototype components), whereas six peaks belonged to the water‐associated herbs group (three metabolites and three prototype components). It was concluded that the serum pharmacochemistry is a meaningful approach for clarifying the difference between blood‐associated and water‐associated herbs in chemical composition. Copyright © 2015 John Wiley & Sons, Ltd.     

Inhibitory effects of tetramethylpyrazine and ferulic acid on spontaneous movement of rat uterus in situ

Tetramethylpyrazine is one of the alkaloids contained in Ligusticum wallichii Franch. Ferulic acid is a phenolic compound contained in Ligusticum wallichii Franch and Angelica sinensis (Oliv.) Diels. The present study was carried out to examine the effect of tetramethylpyrazine and ferulic acid and the combined effect of both compounds on spontaneous uterine contractions in rats in situ. Tetramethylpyrazine and ferulic acid showed an inhibitory effect on uterine movement when given perorally and intravenously, respectively. The combination of both compounds, at doses individually insufficient to inhibit, synergistically inhibited uterine contraction. It was found that tetramethylpyrazine and ferulic acid inhibited uterine contractions and the inhibitory effect induced by the combination of both was due to the potentiation.