基于高分辨质谱物质基础的蓝芩颗粒指纹图谱质量评价研究

目的 建立蓝芩颗粒的HPLC指纹图谱，整体评价该制剂质量。方法 采用HPLC测定蓝芩颗粒的指纹图谱，采用Waters Xselect HSS T3 C₁₈色谱柱(4.6 mm×150 mm，3.5 μm)，乙腈为流动相A，0.05 mol·L^-1甲酸铵的0.05%甲酸溶液为流动相B，梯度洗脱，流速0.7 mL·min^-1，柱温为35℃，检测波长为254 nm；采用HPLC-Orbitrap-MS/MS对蓝芩颗粒物质基础进行研究并对各色谱峰进行归属；同时应用中药色谱指纹图谱相似度软件，对2家生产企业的54批样品进行相似度评价。结果 建立了蓝芩颗粒的HPLC指纹图谱；鉴定共有峰30个，分别归属于板蓝根、黄芩、栀子、黄柏和胖大海5味药；54批样品的相似度为0.92~0.99，相似度评价结果显示，蓝芩颗粒各企业之间的差异较小，整体质量较高。结论 建立的指纹图谱能快速、特征性地对蓝芩颗粒质量进行综合评价。     

鼻渊净胶囊的HPLC指纹图谱研究

目的 建立鼻渊净胶囊的高效液相色谱（HPLC）指纹图谱。方法 采用Agilent SB-C₁₈（4.6 mm×250 mm,5 μm）色谱柱,乙腈-水为流动相、以1.0 ml/min流速行梯度洗脱,检测波长210 nm,柱温30 ℃,洗脱时间为80 min。采用中药色谱指纹图谱相似度评价系统（2004A版）对检测出色谱进行指纹图谱相似度评价。结果 建立了鼻渊净胶囊的HPLC指纹图谱,确定了20个共有峰,15个峰归属到各药材,其中5个峰确认了化学成分;10批样品的指纹图谱的整体相似度与对照图谱比较,均在90%以上。结论 所建立的鼻渊净胶囊指纹图谱有助于从整体上控制该制剂的质量。     

九味羌活颗粒HPLC特征指纹图谱研究

摘 要 目的：建立九味羌活颗粒的HPLC特征指纹图谱,为九味羌活颗粒的质量控制提供依据。方法: 采用Sunfire C₁₈色谱柱(250 mm×4.6 mm,5 μm) ;乙腈(A) 0.1%磷酸(B)为流动相,流速：1.0 ml·min^-1,进行梯度洗脱;检测波长;270 nm,柱温:30℃。结果: 建立了九味羌活颗粒HPLC指纹图谱的共有模式,标定15个共有峰,利用对照品指认4个峰,12批九味羌活颗粒样品HPLC 图谱相似度均大于0.90。结论:所建立的HPLC指纹图谱分析方法简单、重现性良好,可为九味羌活颗粒的品种鉴别及质量评价提供依据。     

杨树花高效液相色谱指纹图谱研究

目的 建立杨树花HPLC指纹图谱以控制杨树花质量。方法 采用HPLC对10批不同产地的杨树花进行指纹图谱构建和方法学考察,运用指纹图谱参数共有峰和相似度进行分析,并对图谱进行聚类分析和主成分分析。结果 建立了杨树花HPLC指纹图谱评价方法,确定了Unitary C₁₈（4.6 mm×250 mm,5µm）色谱柱和乙腈-0.1%甲酸为流动相洗脱系统,在290 nm检测波长下优化了梯度洗脱程序,得到了峰形、分离度较理想的色谱图。通过杨树花HPLC指纹图谱的构建,得到了12个色谱峰,10批杨树花的相似度均在0.9~1.0之间,聚成2类。结论 经方法学考察和统计分析,建立的杨树花HPLC指纹图谱方法稳定、可行。     

黄芩药材指纹图谱研究

摘 要 目的: 建立院内制剂生产采购黄芩药材的HPLC指纹图谱分析方法,为制剂内在质量评价积累数据。方法: 采用HPLC方法,以SHIMADZU VP-ODS C₁₈色谱柱(150 mm×4.6 mm,5 μm);流动相甲醇-0.2%磷酸水(47∶53)等度洗脱;检测波长为280 nm;柱温为30℃;流量为1.0 ml·min^-1,进样量:5 μl。结果:建立黄芩药材指纹图谱,以5号色谱峰黄芩苷为参照峰,确定10个共有峰,测定了11批样品,样品指纹图谱相似度均大于0.8。结论:从整体上显示了购进的不同批次黄芩药材成分特征变化趋势,建立的HPLC指纹图谱方法为含黄芩的院内制剂质量控制提供有效手段。     

甘草配方颗粒HPLC指纹图谱研究

摘 要 目的： 建立甘草配方颗粒的HPLC指纹图谱。方法: 以甘草酸为参照物,利用高效液相色谱梯度洗脱,测定了12批甘草配方颗粒样品;色谱柱为 Waters SunFire C₁₈（250 mm×4.6 mm,5 μm);流动相: 乙腈( A) -0.1%的磷酸溶液( B),检测波长:237 nm,柱温:30 ℃ ,流速:1.0 ml·min^-1。结果: 通过HPLC指纹图谱分析,标示出甘草配方颗粒21个共有色谱峰,相似度均大于0.97,并确认2个已知峰为甘草苷和甘草酸。结论:建立的甘草配方颗粒的HPLC指纹图谱稳定可靠,操作简便,可为甘草配方颗粒质量控制提供科学依据。     

泰山紫草与新疆紫草的HPLC指纹图谱对比研究

目的 对比研究泰山紫草和新疆紫草指纹图谱及其主要成分含量,评价泰山紫草的药用价值。方法 采用HPLC,色谱柱为Symmetry C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为乙腈-水（70∶30）,流速为1.0 mL·min^-1,柱温为30℃,检测波长为516 nm。应用中药色谱指纹图谱相似度评价系统（2012年版）软件,建立泰山紫草、新疆紫草指纹图谱。结果 建立了泰山紫草、新疆紫草指纹图谱的共有模式,以新疆紫草对照图谱作为参照图谱,进行峰匹配,对比计算泰山紫草指纹图谱的相似度为0.798,泰山紫草中含有萘醌类有效成分,但含量与新疆紫草中存在差异。结论 泰山紫草具有一定的药用价值,但不可完全替代新疆紫草使用。     

指纹图谱结合多成分含量测定的猪苓配方颗粒质量评价

目的 建立猪苓配方颗粒质量评价方法,综合评价不同厂家产品质量均一性和稳定性。方法 采用HPLC以Shim-pack GIST C₁₈-AQ （4.6 mm×150 mm,3 μm）为色谱柱,乙腈-水溶液为流动相,梯度洗脱,流速1.0 mL·min^–1,检测波长为350 nm （0~3 min）和250 nm （3~35 min）,柱温30℃,构建不同厂家猪苓配方颗粒指纹图谱,指认共有峰,并进行相似度评价、聚类分析;采用相同HPLC测定4种活性成分含量,对16批样品进行质量分析和评价。结果 猪苓配方颗粒指纹图谱共标定14个共有峰,指认出其中6个成分,分别为2号峰（尿苷）、4号峰（鸟苷）、6号峰（腺苷）、12号峰（猪苓酮B）、13号峰（猪苓酮A）、14号峰（猪苓酮C）;16批样品指纹图谱相似度为0.609~0.982;聚类分析将全部样品分为2大类;鸟苷、腺苷、猪苓酮B、猪苓酮A在各自质量浓度范围内线性关系良好,r均≥0.999 7;仪器精密度、重复性、稳定性试验的RSD均<3％;平均加样回收率分别为98.22%,99.32%,99.56%,99.15%,RSD均<3%（n=6）。16批样品中,鸟苷、腺苷、猪苓酮B、猪苓酮A含量分别为6.326~28.006,13.392~44.058,10.324~30.335,9.270~26.964 μg·g^–1。结论 不同厂家样品存在较大的质量差异,本研究建立的指纹图谱结合多指标性成分含量测定方法可全面、准确评价猪苓配方颗粒内在质量,为整体提升该药品质量提供依据。     

酸枣叶HPLC指纹图谱研究

目的 建立酸枣叶药材的HPLC指纹图谱,为其质量标准的研究提供依据。方法 采用Kinetex C₁₈色谱柱（100 mm×2.1 mm,2.6 μm）;流动相为乙腈-0.1%磷酸酸水溶液,梯度洗脱;体积流量为300 μL/min;柱温30℃,检测波长225 nm,进样量10 μL,对12批酸枣叶药材进行了指纹图谱研究,采用中药色谱指纹图谱相似度评价系统（2012版）软件进行分析。结果 12批酸枣叶的HPLC指纹图谱有13个共有峰,其中1个共有峰得到确认,相似度均＞0.85。结论 该方法准确可靠,重复性好,为更好地控制酸枣叶内在质量提供科学依据。     

岭南特色饮片熟党参HPLC指纹图谱及其炮制前后对比研究

目的 建立同时适用于岭南特色饮片熟党参及其生品的HPLC指纹图谱，分析对比党参蒸制前后成分的变化，为进一步建立熟党参质量控制标准奠定基础。方法 Waters Symmtry C₁₈反相色谱柱（250 mm×4.6 mm，5 μm）；流动相为甲醇-0.1%甲酸水溶液，梯度洗脱（柱温30℃，流速1.0 mL·min^-1，检测波长283 nm，记录时间55 min）。使用中药色谱指纹图谱相似度评价系统（2012版）进行数据处理。结果 不同批次熟党参样品的指纹图谱相似度为0.750~0.979，存在一定的差异性，结合生、熟品色谱峰差异特点选定10个共有特征峰。结论 所建立的方法稳定、重现性好，熟党参与生品之间指纹图谱的差异对比结果为建立熟党参质控标准提供了参考依据。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.