不同产地水飞蓟药材HPLC指纹图谱研究

目的建立水飞蓟药材的HPLC指纹图谱分析方法,并对不同产地水飞蓟药材进行指纹图谱比较研究。方法采用反相高效液相色谱法,Shimadzu C18(150 mm×4.6 mm)为色谱柱,流动相为甲醇-水-冰醋酸(48:52:1),流速:1.0 mL/min,检测波长为287 nm,柱温40℃,进样量10μl。对水飞蓟药材的指标成分水飞蓟宾进行了定性定量,并以相似度鉴别药材质量。结果建立了水飞蓟药材的HPLC指纹图谱,标定了12个共有峰,方法学考察结果符合指纹图谱技术要求,经相似度计算11个不同产地药材之间的相似性均高于0.944。结论本法简便准确、结论可靠,可用于不同产地水飞蓟药材的指纹图谱测定和质量评价,为有效控制水飞蓟药材的内在质量提供了实验依据。     

紫花地丁HPLC指纹图谱研究

目的:建立紫花地丁药材HPLC指纹图谱,提供药材质量控制的可靠方法。方法:采用HPLC方法,以Agi-lent C18(4.6 mm×250 mm,5μm)为色谱柱,甲醇-0.5%醋酸水溶液进行梯度洗脱;检测波长353 nm,流速1.0mL/min。结果:检测了12批不同来源的紫花地丁药材,确立了18个共有峰,建立了紫花地丁对照指纹图谱,计算各被测样品的HPLC指纹图谱的整体相似度,并指认了菊苣苷、七叶内酯、东莨菪素、早开堇菜苷4个特征峰,比较了上述成分在不同药材中的含量。结论:所建立的指纹图谱具有良好的精密度、重现性和稳定性,可作为紫花地丁药材质量控制标准。     

猫须草HPLC指纹图谱研究

为建立猫须草药材HPLC指纹图谱分析方法,采用高效液相色谱法,以Phenomenex Synergi 4u hydro-RP 250×4.60 mm为色谱柱,以甲醇-0.1%甲酸溶液为流动相梯度洗脱,检测波长254 nm,流速1.0 mL·min^-1,柱温40 ℃。结果表明,建立的猫须草药材HPLC指纹图谱,确定了15个共有峰,各猫须草样品指纹图谱与对照指纹图谱的相似度均在0.9以上。该方法简单、准确、重复性好,为更好地控制猫须草药材质量提供有效可靠的方法。     

柘荣太子参HPLC指纹图谱的研究

建立太子参的 HPLC指纹图谱分析条件,为太子参药材内在质量评价积累数据.方法：应用RP-HPLC法;Cosmosil C18分析柱;乙腈-水二元梯度洗脱;流速为1.0 mL/in;检测波长203 nm;分析时间60 min.结果：建立太子参药材指纹图谱,特征共有峰有15个.结论：该方法准确可靠,重复性好,可用于太子参的HPLC的指纹图谱分析.     

中江丹参药材指纹图谱研究

建立四川中江丹参药材的指纹图谱,以期完成中江丹参GAP重要的一环;HPLC法建立丹参脂溶性提取物的指纹图谱;10批中江丹参材指纹图谱符合国家药品监督管理局2000年颁发的关于中药材指纹图谱技术要求;该图谱可作为中江丹参药材质量控制标准之一。     

牛蒡子HPLC指纹图谱的研究

对不同市售地的牛蒡子药材进行指纹图谱的比较研究,以期建立牛蒡子药材的质量评价方法;采用HPLC法,建立了11个不同市售地牛蒡子药材样品的指纹图谱;方法学考察表明,本研究建立的分析方法有较好的重现性,不同市售地牛蒡子药材共有峰的峰面积比有一定的差异;HPLC指纹图谱分析方法可用于评价牛蒡子药材的内在质量。     

叶下珠药材HPLC指纹图谱的比较研究

采用反相高效液相色谱法,对叶下珠药材不同产地、不同采收期、不同部位指纹图谱进行测定和比较分析,并与中成药叶下珠胶囊的HPLC指纹图谱进行了比较研究。结果显示:(1)建立了叶下珠药材的HPLC特征指纹图谱,标定18个共有峰,利用对照品指认4个峰;11批(来源地不同)叶下珠样品的HPLC图谱相似度(相合系数,均值)在0.89~0.99之间。(2)7批不同采收期的叶下珠药材的HPLC指纹图谱相似度在0.94以上,各共有峰的峰面积大多随生长期而增加,至10月5日达到最高,建议叶下珠药材应于每年的10月上旬进行采收。(3)不同部位叶下珠药材HPLC指纹图谱相似度分析发现,叶、果的相似度较高(0.98~0.99),根、茎的相似度较低(0.86~0.87),说明根、茎中各成分含量较低,建议采收叶下珠药材的地上部分即可。(4)比较叶下珠胶囊和叶下珠药材的指纹图谱,发现二者的化学成分非常相似,但峰面积差异较大,其差异可能是由于加工过程所致。该研究所建立的HPLC指纹图谱分析方法简便、重现性好,可用于叶下珠药材及中成药的鉴定与质量评价。     

头花蓼和头状蓼的HPLC指纹图谱鉴定研究

建立头花蓼、头状蓼HPLC指纹图谱鉴定方法。用RP—HPLC（DAD）法,梯度洗脱,测定不同产地头花蓼及头状蓼的指纹图谱,并作相似度比较分析。不同产地头花蓼指纹图谱相似度较好,而头花蓼与头状蓼的指纹图谱有明显差异。HPLC指纹图谱具有重现性好,特征性强,方法简便等特点,可用于头花蓼、头状蓼药材的鉴别。     

吉林12种蒲公英HPLC指纹图谱研究

通过比较吉林产不同种蒲公英HPLC指纹图谱,探讨吉林产蒲公英HPLC指纹图谱的特点,为蒲公英药材的质量控制提供理论参考。采用HPLC方法,以Agilent Extend-C18(250 mm×4.6 mm,5μm)为色谱柱,以甲醇-0.5%冰醋酸水溶液梯度洗脱;检测波长为323 nm,流速为1.000 mL·min-1,柱温35℃,进样量为20μL,检测了12种吉林产不同种类蒲公英药材,确立了9个共有峰,建立了蒲公英对照指纹图谱,计算各被测样品的HPLC指纹图谱的整体相似度,并指认了绿原酸、咖啡酸、总黄酮3个特征峰,比较了上述成分在不同药材中的含量,定量结果表明,3种成分平均含量分别为0.027%、0.026%、0.128%。所建立的指纹图谱具有良好的精密度、重现性和稳定性,可作为吉林产蒲公英药材的质量控制标准。     

菟丝子指纹图谱研究新进展

本研究旨在建立菟丝子药材的高效液相色谱（HPLC）指纹图谱．为科学评价及有效控制菟丝子药材质量提供可靠方法。采用高效液相色谱法,以乙醇提取加水提取作为提取方法制备样品。分析了10批不同采摘时间的菟丝子药材,建立了菟丝子药材的HPLC指纹图谱,确立了10个共有峰,并且利用“中药色谱指纹图谱相似度评价软件”对不同样品之间的相似度进行计算。结果表明该方法准确可靠．重复性好,可为菟丝子药材的质量控制提供科学准确的依据。     

太子参商品药材及其四倍体植株块根的高效液相指纹图谱

对不同产地太子参〔Pseudostellaria heterophylla(Miq.)Pax〕商品药材及其四倍体植株的块根进行了高效液相指纹图谱分析。结果表明,10个批次的不同产地太子参商品药材与经选育获得的6个株系太子参同源四倍体植株块根的HPLC-UV指纹图谱相似度较高,均在0.9以上。选取15个特征峰并大致判断其峰位和比例关系,构成太子参特有的HPLC色谱指纹图谱,为太子参药材鉴别、品质评价及优良品种的选育提供可靠依据。     

Amplified fragment length polymorphism (AFLP) fingerprinting of symbiotic fungi cultured by the fungus-growing ant Cyphomyrmex minutus

A PCR-based fingerprinting technique based on amplified fragment length polymorphisms (AFLP) is used to screen symbiotic fungi of the fungus-growing ant Cyphomyrmex minutus for genetic differences. AFLP fingerprints reveal several fungal ‘types’ that (a) represent distinct clones propagated vegetatively by the ant, or (b) correspond to free-living fungi that may be acquired by the ant. Fungal types identified by AFLP fingerprints correspond to vegetative-compatibility groups established previously, suggesting that vegetative compatibility can be used as a crude indicator of genetic differences between fungi of C. minutus.     

四种中药注射液对大鼠骨髓间充质干细胞增殖作用的影响

目的寻找对骨髓间充质干细胞(MSC)增殖有促进作用的中药。方法通过四甲基偶氮唑盐(MTT)法,Brdu标记MSC,免疫细胞化学法分析4种中药(参麦、生脉、黄芪、刺五加)对大鼠第五代MSC的增殖作用。结果4种中药中除黄芪外都有一定的诱导MSCs增殖的功能,但是刺五加的作用最强。结论刺五加具有较强的促进MSCs增殖的功能。     

Peptide dynamic fingerprints: a tool for investigating the role of conformational flexibility for GLP-1 analogs affinity.

Glucagon-like peptide-1 (GLP-1) is a 30-residue peptide implicated in short-term appetite regulation. Its analogs are presumed to be potential drugs against obesity and non-insulin dependent diabetes mellitus (NIDDM or type 2 diabetes). This study examined how the dynamic fingerprints can be used for establishing dynamics-activity relationships in a series of peptides for which the mechanism of action is unknown and in which mutations can cause an increase or decrease in biological activity. The 3D autocorrelation method was used to generate maps of both active and inactive analogs. As the active conformation of GLP-1 is not yet clearly defined, the dynamic fingerprints of peptides in an aqueous environment were compared to explain the high affinity of the peptide for its receptor. The suggestion that the peptide could bind to the receptor in a folded conformation has been examined. In the case of the GLP-1 analogs, it was shown that the folding tendency cannot be directly related to affinity values and the results do not favor a folded active conformation model of GLP-1.     

Off-rate screening for selection of high-affinity anti-drug antibodies

The rapidly increasing number of therapeutic antibodies in clinical development and on the market requires corresponding detection reagents for monitoring the concentration of these drugs in patient samples and as positive controls for measurement of anti-drug antibodies. Phage display of large recombinant antibody libraries has been shown to enable the rapid development of fully human anti-idiotypic antibodies binding specifically to antibody drugs, since the in vitro panning approach allows for incorporation of suitable blockers to drive selection toward the paratope of the drug. A typical bottleneck in antibody generation projects is ranking of the many candidates obtained after panning on the basis of antibody binding strength. Ideally, such method will work without prior labeling of antigens and with crude bacterial lysates. We developed an off-rate screening method of crude Escherichia coli lysates containing monovalent Fab fragments obtained after phage display of the HuCAL PLATINUM® antibody library. We used the antibody drugs trastuzumab and cetuximab as antigen examples. Using the Octet® RED384 label-free sensor instrument we show that antibody off rates can be reliably determined in crude bacterial lysates with high throughput. We also demonstrate that the method can be applied to screening for high-affinity antibodies typically obtained after affinity maturation.     

Antimutagenicity of extracts from crude drugs in Chinese medicines

The antimutagenicity of extracts from crude drugs was studied by the Ames bioassay system. The crude drugs chosen were medical plants used very frequently as Chinese medicines. Each crude drug was extracted with hot water similar to the method of Chinese medical treatment. Antimutagenicity of the extract was found with 4 kinds of crude drugs, Paeoniae radix, Bupleuri radix, Hoelen and Glycyrrhizae radix. Each extract of the crude drug showed a different type of antimutagenic action from the others.     

Identification of microorganisms using random primed PCR

The polymerase chain reaction has facilitated the use of molecular approaches in microbiology including new strategies for the rapid identification of micro-organisms. Approaches based on the use of random primers and standard conditions, allows characteristic DNA fingerprints to be generated from any micro-organism even in the absence of information about its DNA sequence. Different primers can be used to produce genus-specific, species-specific, or even strain-specific DNA fingerprints. This article covers the background to this strategy, describes three different approaches to generating DNA fingerprints using random primers, and provides experimental detail for one method, RAPD.     

Assessment of 1H NMR spectroscopy and multivariate analysis as a technique for metabolite fingerprinting of Arabidopsis thaliana

An approach to metabolite fingerprinting of crude plant extracts that utilizes 1H nuclear magnetic resonance (NMR) spectroscopy and multivariate statistics has been tested. Using ecotypes of Arabidopsis thaliana as experimental material, a method has been developed for the rapid analysis of unfractionated polar plant extracts, enabling the creation of reproducible metabolite fingerprints. These fingerprints could be readily stored and compared by a variety of chemometric methods. Comparison by principal component analysis using SIMCA-P allowed the generation of residual NMR spectra of the compounds that contributed significantly to the differences between samples. From these plots, conclusions were drawn with respect to the identity and relative levels of metabolites differing between samples.     

陕西秦艽药源植物及秦艽药材HPLC的比较研究

目的:系统比较研究秦艽药材与其药原植物间主要化学成分的差异。方法:高效液相色谱法,色谱柱:Diamonsil C18(200 mm×4.6 mm,5um);洗脱条件:乙腈-水溶液梯度洗脱;检测波长:240 nm;柱温:30℃;流速:0.5 mL.min-1。结果:陇县、太白秦艽中龙胆苦苷含量明显高于市售秦艽药材;通过确定7个共有峰建立陕西秦艽HPLC指纹图谱,所测定的29批秦艽药材具有很高的相似度,而秦艽伪品相似度较低。采用HPLC各峰面积及其相对之比进行聚类分析结果显示,采自太白的10个样本和陇县的8个样本分别聚为两支,表明秦艽化学成分含量与其地理分布存在密切关系。结论:本实验建立的指纹图谱可用于秦艽及其药材鉴别和质量控制;聚类分析可以鉴别秦艽产地。     

滁菊的HPLC指纹图谱制作及其化学计量学分析

建立滁菊的HPLC指纹图谱,结合化学计量学手段对不同滁菊的指纹图谱进行分析研究,以期为滁菊的质量控制和产地追溯提供依据。采用HPLC法建立指纹图谱,并用中药色谱指纹图谱相似度评价系统(2012版)和系统聚类、主成分分析2种化学计量学法对指纹图谱和特征峰进行分析。分析结果发现样品有27个共有峰,12个滁菊样品具有较高的相似度,杭菊与滁菊的相似度较差,聚类分析与主成分分析结果和相似度分析结果一致。将HPLC指纹图谱与化学计量学结合可对滁菊进行鉴别和质量评价,为其质量控制和追溯提供了理论参考。