高效毛细管电泳法测定钩藤中的钩藤碱

目的分离钩藤碱与异钩藤碱,测定钩藤中的钩藤碱。方法采用高效毛细管电泳法,运行缓冲液为0.05 mol.L-1硼砂溶液(pH6.9)-0.02 mol.L-1羟丙基-β-环糊精和0.05 mo.lL-1胆酸钠的甲醇溶液(85:15),操作电压为12 kV,温度为40℃,检测波长为250 nm。结果钩藤碱0.025~0.496 mg.ml-1与峰面积呈良好的线性关系(r=0.9998),加样回收率为99.8%,RSD=1.91%。结论所用方法可快速有效地分离钩藤碱及异钩藤碱,并测定植物中的钩藤碱。     

HPLC法测定安乐片中钩藤碱和异钩藤碱的含量

目的：建立安乐片中钩藤碱和异钩藤碱的含量测定方法。方法：用高效液相色谱法Agilent XDB C₁₈(250 mm×4.6 mm,5μm);甲醇-0.2%氨水溶液(60:40)为流动相;检测波长为245nm,流速为1 mL.min_-1,柱温25 ℃。结果：钩藤碱进样量在0.0285～0.7133ug(r=0.9999)与峰面积线性关系良好;异钩藤碱进样量在0.0166～0.416ug(r=0.9998)范围内线性良好, 钩藤碱的平均回收率为98.61%,RSD为1.56%,异钩藤碱的平均回收率为97.49%,RSD为1.27%。结论：本法灵敏、准确,专属性强,能简便有效的控制安乐片中钩藤碱和异钩藤碱的含量。     

HPLC法测定钩藤中钩藤碱和异钩藤碱含量

目的建立中药钩藤中钩藤碱和异钩藤碱含量的高效液相色谱检测方法,为中药钩藤中有效成分的含量测定提供科学的参考。方法高效液相色谱条件为:色谱柱Phenomenex C18(250nm×4.0nm,5μm),流动相:甲醇-水(55∶45),紫外检测波长:254nm,流速:1.0mL/min,柱温:28℃,进样量:20μL。结果以峰面积为纵坐标浓度为横坐标,计算得到钩藤碱和异钩藤碱回归方程分别为Y=0.999E+6x-21133,R2=0.9998和Y=1.008E+6x-13556,R2=0.9999。结果显示钩藤碱和异钩藤碱分别在0.244～1.534μg和0.226～1.288μg浓度范围内其浓度与峰面积具有良好的线性关系。另外本法在精密度、稳定性、重复性上均取得了较好结果。结论高效液相色谱法测定钩藤中藤碱和异钩藤碱含量,具有快速、简便、重复性好的特点,值得在中药含量测定上大力推广应用。     

钩藤总碱中钩藤碱和异钩藤碱定量方法的建立及稳定性试验

目的建立钩藤总碱中钩藤碱和异构藤碱的含量测定方法并对其稳定性进行研究。方法采用C18柱(4.6 mm×150 mm,5μm),甲醇∶0.1 mol/L三乙胺溶液(冰醋酸调pH 7.5)44∶56为流动相,流速为1.0 ml/min。检测波长254 nm。比较分别放置在室温、6℃条件下的同一批次2份总碱中钩藤碱、异构藤碱的含量变化。结果钩藤碱进样量在0.020 2～0.101 mg/ml线性关系良好,r=0.999 9;异构藤碱进样量在0.020 6～0.103 mg/ml线性关系良好,r=0.999 9,钩藤碱平均回收率为99.67%,RSD为0.97%;异钩藤碱平均回收率为99.95%,RSD为1.02%。结论建立的方法简便、准确,可作为钩藤总碱的含量测定方法;钩藤碱、异构藤碱不稳定,可随时间延长含量下降,且温度对二者皆有影响,对钩藤碱影响较大。     

以离子液体为流动相添加剂的HPLC法分离并测定钩藤中钩藤碱和异钩藤碱的含量

目的:建立测定钩藤中钩藤碱和异钩藤碱含量的方法。方法:以离子液体1-丁基-3-甲基咪唑氯化盐(C_4mimCl)为流动相添加剂,与无添加剂的流动相以及加入传统添加剂三乙胺(对色谱柱有损伤)后对高效液相色谱(HPLC)法分离钩藤中钩藤碱和异钩藤碱的分离度进行比较,筛选C_4mimCl的最佳浓度,用新建立的方法测定江西4个产地钩藤中钩藤碱和异钩藤碱的含量。色谱柱为Dikmatech Diamonsil Plus C_(18),流动相为乙腈-缓冲液(0.1%磷酸+3.0 mmol/L C_4mimCl),梯度洗脱,紫外检测波长为245 nm,流速为1 mL/min,进样量为10μL。结果:当流动相中无添加剂时、加入3.0 mmol/L三乙胺或3.0 mmol/L C_4mimCl作添加剂时,钩藤碱与前峰分离度分别为1.02、1.23、1.72,与后峰分离度分别为1.06、6.00、4.25,对称因子分别为0.81、0.86、1.13;异钩藤碱与前峰分离度分别为0.96、3.89、4.05,与后峰分离度分别为1.02、2.34、2.36,对称因子分别为0.88、0.81、0.96。钩藤碱、异钩藤碱检测质量浓度线性范围分别为4.93～157.76(r=0.999 9)、4.98～159.50μg/m L(r=1.000 0),定量限分别为0.486 4、0.793 6μg/mL,精密度、重复性、稳定性和耐用性试验中的RSD均小于5%(n=6),回收率分别为102.9%～107.8%(RSD=1.7%,n=6)、95.4%～106.3%(RSD=3.9%,n=6)。4个产地钩藤中钩藤碱和异钩藤碱的含量范围分别为0.758～1.343、1.511～1.823 mg/g。结论:C_4mimCl加入到流动相中能提高分离度,且以此建立的HPLC法快速、准确、重复性好,可用于钩藤中钩藤碱和异钩藤碱的含量测定。     

HPLC法测定不同煎剂中钩藤碱和异钩藤碱煎出效率

目的:测定两种不同方剂中钩藤碱与异钩藤碱煎出效率,并得到其最佳煎煮时间.方法:采用HPLC法,色谱柱为C18色谱柱(4.6mm×200mm,5μm):流动相:甲醇-水=68:32(含0.2％三乙胺水溶液,冰乙酸调pH=7.0),流速为0.8mL/min,波长254nm,柱温30℃.结果:钩藤碱和异钩藤碱分别在0.0021～0.05mg/mL(r2=0.9997)和0.0023～0.872mg/mL(r2=0.9998)范围内有良好的线性关系.讨论:钩藤饮片在不同方剂中应用煎法后下时,最佳煎出效率不一致.天麻钩藤饮中11分钟最佳,半夏天麻钩藤汤中13分钟最佳.     

天麻钩藤颗粒中钩藤的鉴别及天麻素的测定

目的 鉴别天麻钩藤颗粒中的钩藤并测定天麻素的含量.方法 采用TLC法鉴别钩藤;HPLC法测定方中天麻的含量.结果 TLC鉴别分离良好,阴性样品无干扰;天麻素进样量0.0181～1.4464μg与峰面积的线性关系良好(r=0.9999),平均回收率为98.0%,RSD=1.41%(n=6).结论 所建方法专属性强、重复性好,可控制中药复方制剂中钩藤与天麻的质量.     

天麻钩藤合剂质量标准的研究

目的建立本院医院制剂天麻钩藤合剂的质量控制方法。方法采用薄层色谱法(TLC)对黄芩、栀子、天麻及忍冬藤进行定性鉴别;采用高效液相色谱法对天麻钩藤合剂中天麻素、黄芩苷、绿原酸和栀子苷4种成分进行含量测定。结果 TLC能检验出黄芩、栀子、天麻和忍冬藤,且具有特征性的斑点。黄芩苷在0.018 00~0.180 00 mg与峰面积线性关系良好,平均加样回收率为100.0%,RSD=0.70%。天麻素在0.000 60~0.006 00 mg与峰面积线性关系良好,平均加样回收率为99.9%,RSD=1.3%。绿原酸在0.000 36~0.003 60 mg与峰面积线性关系良好,平均加样回收率为100.4%,RSD=1.8%。栀子苷在0.005 00~0.050 00 mg与峰面积线性关系良好,平均加样回收率为100.8%,RSD=1.7%。结论所建立的方法准确性高、灵敏度和重复性好,可作为天麻钩藤合剂的质量控制标准。     

HPLC测定黔产钩藤不同部位钩藤碱和异钩藤碱的含量

目的 HPLC测定黔产钩藤不同部位中钩藤碱和异钩藤碱的含量,并比较其差异。方法 以钩藤碱、异钩藤碱为对照品,采用CAPCELL PAK C₁₈色谱柱（4.6 mm×250 mm,5μm）,以乙腈-0.1%三乙胺水溶液（1%磷酸调pH=9）为流动相,流速1 mL·min^-1,检测波长245 nm,柱温30℃,进样量10µL。结果 钩藤碱与异钩藤碱分离良好,线性、重复性以及回收率均符合要求,钩藤不同部位中的含量存在差异,由高到低依次为皮 > 茎 > 带钩茎枝 > 叶,而茎薄壁组织与木质部未检测到钩藤碱与异钩藤碱。结论 该含量测定结果为钩藤资源的充分利用提供了科学依据。     

复方附子口服液中黄芪甲苷含量测定和乌头碱限量检查方法研究

目的建立复方附子口服液中黄芪甲苷含量测定和乌头碱限量检测方法。方法采用HPLC法对黄芪甲苷含量、乌头碱限量进行检测。结果黄芪甲苷在0.047 12⁰.471 2 mg·mL-1与峰面积线性关系良好(r=0.999 7),高、中、低浓度平均加样回收率分别为99.9%、99.9%、97.6%,RSD分别为4.4%、1.9%、2.2%;乌头碱在0.005 970.471 2 mg·mL-1与峰面积线性关系良好(r=0.999 7),高、中、低浓度平均加样回收率分别为99.9%、99.9%、97.6%,RSD分别为4.4%、1.9%、2.2%;乌头碱在0.005 97⁰.597 0 mg·mL-1与峰面积线性关系良好(r=0.999 9);高、中、低浓度平均加样回收率分别为101.3%、98.3%、98.7%,RSD分别为2.1%、1.0%、1.6%。结论该方法专属性强,灵敏度高,重现性好,可用于复方附子口服液中黄芪甲苷含量和乌头碱限量的检测。     

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.

---

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.     

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.