混合取代基—β—环糊精—毛细管电泳法分离测定麻黄碱和伪麻黄碱

目的:建立毛细管电泳法同时分离测定同分异构的2对手性化合物麻黄碱和伪麻黄碱的含量。方法:Waters CapillaryIon Analyzer,50μm×60cm空心熔融石英毛细管柱,柱上紫外检测。电泳条件:分离用缓冲液为25mmol·L~(-1)的磷酸-Tris 缓冲液,含18mmol·L~(-1)的羧甲基-β-环糊精和7.5mmol·L~(-1)的β-环糊精-硫酸酯,pH3.02。分离电压:18kV;温度:25℃;虹吸进样,高度10cm,时间1s;紫外检测波长214nm。结果:左旋伪麻黄碱、右旋麻黄碱、左旋麻黄碱和右旋伪麻黄碱线性范围分别为23.7～237μg·ml~(-1),25.0～250μg·mL~(-1),25.0～250μg·mL~(-1),25.4～254μg·mL~(-1),以信噪比等于3:1为标准,最小检测浓度均为5.0μg·mL~(-1);日内和日间精密度RSD在2.4％～4.3％之间。结论:方法分离效率高、简便、快速、成本低。     

RP—HPLC法测定益列康宁片中美帕曲星的含量

目的:采用RP-HPLC法测定益列康宁片中美帕曲星的含量。方法:采用LiChroCART RP-18不锈钢柱,以乙腈和0.005 mol·L~(-1)乙二胺四乙酸二钠混合溶剂作流动相,梯度洗脱,流速1.0 mL·min~(-1),检测波长378 nm。结果:美帕曲星在100～300μg·mL~(-1)浓度范围内呈现良好的线性关系(r=0.999 9),平均回收率为99.4％～100.4％,RSD<1.3％(n=3)。日内RSD为0.5％～0.9％(n=5),日间RSD为1.2％～1.7％(n=5)。结论:本法简便,精密度高,重现性好,结果准确可靠。     

高效液相色谱法测定莫达非尼及其胶囊剂含量

目的:测定莫达非尼原料药及其胶囊剂的含量。方法:采用高效液相色谱法。色谱柱为Phenomenex ODS(150mm×4.6mm,5μm),柱温30℃,流动相为甲醇-50mmoL·L~(-1)甲酸(50:50,pH=2.5),流速1.0mL·min~(-1),UV检测波长225nm。结果:莫达非尼浓度在1.0～20μg·mL~(-1)范围标准曲线线性良好,最低检测浓度0.5μg·L~(-1),日内RSD为1.8％(n=5),日间RSD为2.0％(n=5),加样回收率98.0％～99.6％(n=3)。结论:本法简便快速,准确可靠,可用于莫达非尼及其胶囊剂含量测定等相关研究。     

HPLC法测定人血浆中盐酸二甲双胍浓度

目的:建立血浆中盐酸二甲双胍浓度测定方法。方法:血浆在酸性条件下以乙腈沉淀蛋白后,用二氯甲烷萃取纯化,以乙腈-5 mmol·L~(-1)磷酸盐缓冲液(pH 4．8)(45:55,每100 mL流动相中含十二烷基硫酸钠0．17 g)为流动相,色谱柱为DIAMONSIL~(TM)C_(18)柱(250 mm×4．6 mm,5μm),检测波长为233 nm,柱温为40℃。流速为1．2 mL·min~(-1)。结果:在此色谱条件下,盐酸二甲双胍与血浆中其他成分分离完全,线性范围为60．06～4 004 ng·mL~(-1),最低检测浓度为60．06 ng·mL~(-1),相对回收率在101．3％～108．3％,日内精密度RSD＜5．4％,日间精密度RSD＜13％。结论:该法稳定、灵敏、可靠,可用于人体血浆中盐酸二甲双胍浓度的测定。     

复方石韦片中生物碱的分离测定

目的:建立了复方石韦片中槐果碱和苦参碱的分离测定方法。方法:采用高效毛细管电泳法进行测定。运行电压27kV;运行缓冲液为0.2 mol·L~(-1) Tris-40 mmol·L~(-1)磷酸二氢钠-5％异丙醇,磷酸调pH 5.75;进样方式:真空进样;进样时间:5s;检测波长:205nm。结果:槐果碱和苦参碱的线性范围分别为0.5～5.76μg·mL~(-1)和0.66～13.2μg·mL~(-1),相关系数分别为0.9991和0.999 3,平均回收率分别为99.5％和99.3%,RSD分别小于4％和2％(n=3)。结论;本法简便、快速、重现性好。     

毛细管区带电泳法测定马钱子药材及其制剂中士的宁和马钱子碱的含量

目的:建立了测定马钱子药材及其制剂中土的宁和马钱子碱含量的毛细管区带电泳法。方法:选择25mmol·L~(-1)醋酸钠含0.01％冰醋酸(pH3.86)溶液为运行缓冲液,无涂层毛细管柱(57cm×75μm,有效长度50cm),采用5kPa,5s压力进样,运行电压30.0 kV,温度20℃,DAD检测波长203nm。实验以盐酸麻黄碱为内标物质。结果:士的宁和马钱子碱的浓度分别在6.93～138.20μg·mL~(-1)和6.51～130.20μg·mL~(-1)范围内时,具有良好的线性关系,相关系数r分别为0.9997和0.9994(n=6);马钱子药材中士的宁和马钱子碱的加样回收率分别为101.3％和99.42％;RSD分别为2.5％和2.2％(n=6)。士的宁和马钱子碱的最低检测限分别为0.05μg·mL~(-1)和0.06μg·mL~(-1)。结论:马钱子药材及其制剂中的其它组分与士的宁和马钱子碱可产生良好分离,本法简便、快速、准确、可靠。     

反相高效液相色谱法测定多糖包覆脂质体中胰岛素

目的:建立反相高效液相色谱法测定多糖包覆的胰岛素脂质体含量和包封率。方法:采用逆相蒸发-超声法制备胰岛素脂质体;用高效液相色谱法和超速离心法测定胰岛素脂质体含量与包封率;采用反相色谱柱Lichrospher ODS-C_(18)(250 mm×4.6 mm,5μm),以0.025 mol·L~(-1)硫酸盐缓冲液(pH 2.0)-己腈(72:28)为流动相,流速1.0 mL·min~(-1),柱温40℃,检测波长214nm。结果:线性范围为2～10 μg·mL~(-1)。在空白壳聚糖包覆脂质体中胰岛素加样回收率为94.65％～100.5％,日内、日间精密度的RSD分别为0.92％～1.6％和4.0％～5.6％。在空白海藻酸钠包覆脂质体中胰岛素加样回收率为96.12％～101.1％,日内、日间精密度的RSD分别为1.7％～2.4％和4.0％～4.7％。壳聚糖包覆的胰岛素脂质体和海藻酸钠包覆的胰岛素脂质体包封率分别为73.6％和68.7％,胰岛素总含量分别为0.682 mg·mL~(-1)和0.713mg·mL~(-1)。结论:本方法便捷、灵敏、准确、重现性好,可用于测定多糖包覆的胰岛素脂质体含量与包封率。     

反相高效液相色谱法测定利福昔明的含量

目的:采用反相高效液相色谱法测定利福昔明的含量。方法:色谱柱为C_(18)柱,流动相为甲醇—0.075mol·L~(-1)KH_2PO_4溶液-1.0mol·L~(-1)枸橼酸溶液(75:26:4),检测波长为254nm;流速为1.0mL·min~(-1)。结果:线性范围:10.0～100.0μg·mL~(-1),r=0.9999,回收率99.2％。供试品溶液在8h内稳定性良好,日内重复性RSD=0.24％,日间重复性RSD=0.50％。结论:本方法简单、快速,结果准确。     

高效液相色谱法测定脂质体中盐酸柔红霉素的含量

目的:测定脂质体中盐酸柔红霉素的含量。方法:以C_(18)色谱柱为分析柱,乙腈-0.02 mol·L~(-1)磷酸二氢钠溶液-三乙胺(34:66:0.3,磷酸调pH为4.0)为流动相,流速1.0 mL·min~(-1),于检测波长233nm下进行检测,进样量50μL。结果:在2.51～25.10μg·mL~(-1)范围内,盐酸柔红霉素对照品的峰面积(A)与其浓度(C)呈现良好的线性关系(r=0.9993)。最低检测限40ng·mL~(-1)。低、中、高3种浓度平均回收率为100.8％,103.4％,102.2％(n=5); RSD分别为0.49%,0.42％,1.9％。日内、日间RSD(n=5)分别为1.1％～1.6％和0.60％～1.5％。结论:本法用于测定脂质体中盐酸柔红霉素的含量快速、简便、专属性较高。     

HPLC法测定人血浆中劳拉西泮的浓度

目的:建立测定人血浆中劳拉西泮浓度的HPLC法。方法:采用美国Dikma公司Diamonsil~(TM)C_(18)(250mm×4.6mm,5μm)为色谱柱,流动相为甲醇-水(75:25,V/V),流速为0.8mL·min~(-1),检测波长232mm,以乙酸乙酯为提取溶剂。结果:劳拉西泮高(20.00μg·mL~(-1))、中(6.00μg·mL~(-1))、低(0.60μg·mL~(-1))3个浓度的平均回收率分别为101.67％,98.17％,91.20％,日内、日间RSD均<5％(n=5);分析方法的定量测定下限为0.O05μg·mL~(-1)。线性范围为0.01～20.00μg·mL~(-1),回归方程为Y=2.16×10~(-2)X-3.33×10~(-3),r=0.9999(n=10)。结论:该方法灵敏、准确、简单、快速,可用于劳拉西泮临床血药浓度监测和药动学研究。     

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.

     

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.     

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.     

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.