紫外分光光度法和高效液相色谱法测定淫羊藿总黄酮含量的比较研究

目的:比较紫外分光光度法和高效液相色谱法测定淫羊藿总黄酮含量的差异,探讨淫羊藿总黄酮含量测定方法的可靠性。方法:紫外分光光度法以淫羊藿苷为指标性成分,在270 nm 波长处进行测定。高效液相色谱法以 ZORBAX SB-C_(18)柱(250mm×4.6 mm,5μm)为分析柱,乙腈-水梯度洗脱,流速1.0 mL·min~(-1),检测波长为270 nm。经紫外光谱识别的黄酮类成分(部分经 ESI-MS 确认),除朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ和宝藿苷Ⅰ以自身对照外,其他黄酮类成分均以淫羊藿苷为参比进行定量,淫羊藿总黄酮含量等于朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ、宝藿苷Ⅰ和其他黄酮成分含量之和。结果:紫外分光光度法,淫羊藿苷在2.45～24.50μg·mL~(-1)范围内线性关系良好(r=0.9996),测得淫羊藿总黄酮含量以淫羊藿苷计为56.6%。高效液相色谱法中,与淫羊藿苷紫外光谱相似的33个色谱峰,MS 归属了其中10个主要成分均为黄酮类成分。朝藿定 C、淫羊藿苷、鼠李糖基淫羊藿次苷-Ⅱ和宝藿苷Ⅰ分别在0.093～1.852μg(r=0.9998)、0.107～2.136μg(r=0.9997)、0.094～1.876μg(r=0.9998)、0.098～1.956μg(r=0.9998)线性关系良好,淫羊藿总黄酮含量为35.6%。结论:紫外分光光度法和高效液相色谱法测定的淫羊藿总黄酮含量差异显著,紫外分光光度法的准确性有待进一步考证。     

RP-HPLC测定淫羊藿不同品种和部位中柔藿苷和淫羊藿苷

目的:为了对中药淫羊藿中柔藿苷和淫羊藿苷含量进行系统研究,测定了淫羊藿不同品种和药用部位的柔藿苷和淫羊藿苷含量。方法:采用RP-HPLC法对3个品种多个样品进行测定,以BECKMAN COULTER_(TM)-C_(18)柱(带预柱)为色谱柱,流动相为乙腈-水(25∶75),流速1.0mL·min~(-1),检测波长为270nm。结果:柔藿苷在0.19642-1.9642μg范围内呈良好的线性关系,淫羊藿苷在0.19335-1.9335μg范围内呈良好的线性关系。巫山淫羊藿同一植株体的不同部位中柔藿苷和淫羊藿苷的分布均为叶>根>茎。巫山淫羊藿同一植株的任何部位都是柔藿苷含量高于淫羊藿苷,而淫羊藿和箭叶淫羊藿叶中都是淫羊藿苷含量高于柔藿苷。结论:巫山淫羊藿中以柔藿苷为主要成分,淫羊藿和箭叶淫羊藿中以淫羊藿苷为主要成分。     

不同采收季节的淫羊藿中朝藿定C和淫羊藿苷的含量比较

目的比较3个贵州主流产淫羊藿药材在不同采收季节中朝藿定C和淫羊藿苷的含量。方法用高效液相色谱（HPLC）法测定粗毛淫羊藿、巫山淫羊藿、黔岭淫羊藿中朝藿定c和淫羊藿苷的含量。采用EliteSinoChromODS—AP柱（250mm×4．6mm,5．0Ixm）,流动相为乙腈（A）一水（B）梯度洗脱（0—22rain,27％_29％A;22～23rain,29％_÷100％A;23～34rain,100％A;34～36rain,loO％一27％A;36～50min,27％A）。结果朝藿定c和淫羊藿苷进样量分别在13．22～396．6Ixg（r=0．9999）和3．026～151．3斗g范围内与峰面积均呈良好线性关系（r=0．9999）,平均回收率均在98．0％一105．O％范围内;粗毛淫羊藿中朝藿定c和淫羊藿苷的含量分别为0．516％～2．973％,0．096％～0．216％;黔岭淫羊藿中朝藿定C和淫羊藿苷的含量分别为0．071％～0．185％,0．081％～0．164％;巫山淫羊藿含朝藿定c和淫羊藿苷中朝藿定c和淫羊藿苷的含量分别为1．015％～4．219％,0．080％～0．190％。结论巫山淫羊藿中朝藿定c和淫羊藿苷的含量最高,其次为粗毛淫羊藿、黔岭淫羊藿,说明不同产地、不同品种的淫羊藿药材中朝藿定c和淫羊藿苷的含量差异较大．     

淫羊藿苷转化产物的制备及活性研究

目的旨在分析淫羊藿苷及转化产物抑制α-葡萄糖苷酶活性研究。方法对淫羊藿苷进行酶水解、酸水解、酶水解与酸水解相结合的方式进行结构转化,采用薄层色谱法、理化方法、核磁共振波谱法对转化产物进行结构鉴定,分别得到淫羊藿次苷Ⅰ、淫羊藿次苷Ⅱ、水合淫羊藿素、环淫羊藿素及脱水淫羊藿素。采用pNPG法建立α-葡萄糖苷酶筛选模型,对淫羊藿苷及其产物进行α-葡萄糖苷酶抑制活性筛选。结果体外活性试验果表明,脱水淫羊藿素、淫羊藿次苷Ⅰ及淫羊藿次苷Ⅱ体外活性较淫羊藿苷好,其中脱水淫羊藿素(IC_(50)=0.102 mg/mL)的活性最好,与拜糖平(IC_(50)=0.850 mg/mL)相比抑制活性更强。结论淫羊藿苷转化产物可作为继续进行结构修饰的先导化合物,具有深入研究和开发的前景。     

延年乐口服液含量测定的研究

目的建立高效液相色谱法测定延年乐口服液中淫羊藿苷的含量,以增加对制剂的质量控制。方法采用迪马公司(DIKMA)十八烷基硅烷键合硅胶,4.6×150mm5μm色谱柱,以甲醇-水(65∶35)为流动相,检测波长为270nm。结果淫羊藿苷浓度在18.968~113.808μg.mL-1范围内线性关系良好,r=1.000,平均回收率为97.45%,RSD为1.70%。结论本法测定淫羊藿苷简便、准确、重现性好,可用于延年乐口服液中淫羊藿苷的含量测定。     

RP-HPLC法同时测定不同品种淫羊藿药材中5种主要黄酮类成分的含量

目的:建立以反相高效液相色谱法测定不同品种淫羊藿药材中5种主要黄酮类成分含量的方法。方法:色谱柱为Zorbax SB-C18(250mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),检测波长为270nm,柱温为30℃,同时测定淫羊藿药材中淫羊藿苷、朝藿定A、朝藿定B、朝藿定C、宝藿苷Ⅰ的含量。结果:5种成分均能达到基线分离,线性良好。不同品种淫羊藿药材中5种主要黄酮类成分的含量差别较大。结论:建议在考察淫羊藿药材的质量时,不能只考察淫羊藿苷,应建立多种黄酮类成分测定指标,以全面控制淫羊藿药材的质量。     

全子强身酒质量标准研究

<正>全子强身酒主要由枸杞子、山茱萸、五味子、淫羊藿、女贞子等27味药组成。为有效控制该品种的质量,制订了以下测定方法。1薄层色谱鉴别1.1仪器与试药:全子强身酒(山西全邦医药保健品有限公司);熊果酸、五味子甲素、五味子乙素、淫羊藿苷对照品     

12种中成药中淫羊藿组分的质量考察

目的:考察12种市售不同剂型含淫羊藿中成药中淫羊藿2个指标成分的含量,为有效控制含淫羊藿的中成药质量提供依据。方法:采用高效液相色谱法分别测定市售12种中成药中朝藿定C和淫羊藿苷的含量。色谱柱为Venusil ASB-C18(250mm×4.6mm,5μm),流动相为乙腈-水(梯度洗脱),流速为1.0mL·min-1,检测波长为272nm,柱温为20℃。结果:市售大部分含淫羊藿中成药所使用的淫羊藿原料中淫羊藿苷的含量均达不到药典规定的质量标准。结论:应将朝藿定C作为淫羊藿药材及含淫羊藿中成药的评价指标之一。     

反相高效液相色谱法测定淫羊藿中淫羊藿苷的含量

目的建立用于测定淫羊藿中淫羊藿苷含量的反相高效液相色谱法。方法采用ZORBAX Ecilpse XDB-C18（150mm×4．6mm,5μ）色谱柱;以水-乙腈（74：26,V/V）为流动相;检测波长270nm;流速1．0mL·min^-1,柱温25℃。结果淫羊藿昔质量浓度在5～50mg·L^-1内与其峰面积之间线性关系良好（r=0．9999）,平均加样回收率为100．32％,RSD为0．44％。结论本方法操作简便、快速,结果准确,可用于淫羊藿中淫羊藿苷的含量测定。     

HPLC法同时测定淫羊藿药材中8种黄酮类成分的含量

目的建立HPLC法同时测定淫羊藿药材中朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷I等8种黄酮类成分含量的方法。方法色谱柱为Agilent Zorbax SB-C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-水梯度洗脱,检测波长为270 nm,流速为1.0 m L·min-1,柱温为30℃。结果朝藿定A、朝藿定B、朝藿定C、淫羊藿苷、宝藿苷I、朝藿苷丙、epimedokoreanoside I、朝藿苷乙等8种黄酮类成分的线性关系良好,线性范围分别为2.0²0.2、1.020.2、1.0⁵7.2、3.057.2、3.0¹32.0、1.0132.0、1.0⁵0.0、1.050.0、1.0⁴5.2、0.445.2、0.4¹5.0、0.615.0、0.6²5.8、0.225.8、0.2⁵2.7 mg·L-1;平均回收率分别为98.3%、96.7%、97.2%、96.9%、95.5%、98.5%、98.2%、98.5%,RSD分别为1.0%、0.7%、0.9%、0.6%、0.2%、1.1%、1.4%、0.3%(n=6)。结论该方法可作为淫羊藿药材含量测定的方法。对收集的104批市售的淫羊藿药材中淫羊藿苷含量进行了测定,淫羊藿苷含量质量分数为0.01%52.7 mg·L-1;平均回收率分别为98.3%、96.7%、97.2%、96.9%、95.5%、98.5%、98.2%、98.5%,RSD分别为1.0%、0.7%、0.9%、0.6%、0.2%、1.1%、1.4%、0.3%(n=6)。结论该方法可作为淫羊藿药材含量测定的方法。对收集的104批市售的淫羊藿药材中淫羊藿苷含量进行了测定,淫羊藿苷含量质量分数为0.01%⁰.17%,大部分不符合《中华人民共和国药典》标准。     

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.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

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.