HPLC法测定黄柏不同炮制品中小檗碱的含量

目的考察不同炮制方法对黄柏中小檗碱含量的影响。方法采用高效液相色谱法测定盐酸小檗碱含量。色谱柱ZORBAX SB-C18;流动相为乙腈-0.1%磷酸溶液（50∶50）;流速为1.0mL·min^-1;柱温为室温;检测波长为265nm;进样量为20μL。结果在0.46-4.6μg（r=0.9998,）范围内线性关系良好。结论结果表明黄柏丝的小檗碱含量最高。     

HPLC法同时测定川黄柏中盐酸黄柏碱和盐酸小檗碱的含量

目的:建立同时测定川黄柏中盐酸黄柏碱和盐酸小檗碱含量的方法。方法:采用高效液相色谱法。色谱柱为Waters XTERRA?MS C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液(每100 ml中加0.1 g十二烷基磺酸钠,梯度洗脱),检测波长为284 nm(盐酸黄柏碱)和265 nm(盐酸小檗碱),流速为1.0 ml/min,柱温为30℃。结果:盐酸黄柏碱和盐酸小檗碱的质量浓度分别在9.45²36.15、74.77236.15、74.77¹ 869.25 mg/L范围内与各自峰面积积分值呈良好的线性关系(r分别为0.999 1、0.999 8);二者精密度、稳定性、重复性试验的RSD≤1.55%;平均加样回收率分别为97.47%1 869.25 mg/L范围内与各自峰面积积分值呈良好的线性关系(r分别为0.999 1、0.999 8);二者精密度、稳定性、重复性试验的RSD≤1.55%;平均加样回收率分别为97.47%⁹8.24%、95.03%98.24%、95.03%⁹5.63%,RSD分别为0.39%95.63%,RSD分别为0.39%⁰.93%、0.42%0.93%、0.42%⁰.93%(n均为3)。结论:该方法简便、快速、专属性强,可有效评价川黄柏中盐酸黄柏碱和盐酸小檗碱的质量。     

反相高效液相色谱法测定复方黄柏酊中小檗碱含量

目的建立复方黄柏酊中小檗碱的含量测定方法。方法用中性氧化铝吸附法制备样品,采用反相高效液相色谱法测定含量,色谱柱为C18柱(200mm×4.6mm,5μm),柱温为25℃,流动相为水-乙腈-磷酸二氢钾-十二烷基硫酸钠(50∶50∶0.3∶0.2),流速为1.0mL/min,检测波长为345nm。结果盐酸小檗碱浓度在50～250μg/mL范围内与峰面积线性关系良好,回归方程为A=46680.1C-154.0,r=0.9998(n=5),平均回收率为99.06%,RSD=0.19%。结论该方法灵敏、准确,重现性好,可作为复方黄柏酊中小檗碱的质量控制标准。     

RP-HPLC法同时测定复方黄柏液中盐酸小檗碱、绿原酸和连翘苷的含量

目的采用高效液相色谱梯度洗脱法同时测定复方黄柏液(黄柏、连翘、金银花、蒲公英、蜈蚣)中盐酸小檗碱、绿原酸和连翘苷的含量,为制定该制剂质量标准中含量测定方法及限度提供依据。方法Scienhome(Kromasil C18,5μm,200×4.6mm)色谱柱;流动相为乙腈-0.4%磷酸溶液梯度洗脱(5∶95~25∶75);检测波长:0~40min:327nm;40~63min:277nm;63~70min:265nm;流速1.00mL.min-1;柱温30℃;进样量10μL。结果该方法回收率盐酸小檗碱:100.5%,绿原酸:98.8%,连翘苷:99.9%。结论该方法能够将盐酸小檗碱、绿原酸、连翘苷很好的分离,测定结果准确可靠,重现性好。     

HPLC法测定黄柏炮制前后盐酸小檗碱的含量

目的考察黄柏炮制前后盐酸小檗碱的含量变化。方法色谱柱:Lana C18柱(4.6mm×250mm,5μm,美国Phenom enex公司):流动相:甲醇-乙腈-水(42∶16∶42)(含0.05mol.L-1十二烷基硫酸钠及0.037mol.L-1酒石酸);检测波长:345nm。结果盐酸小檗碱在0.1108~1.108μg范围内,浓度与峰面积线性关系良好(r=0.9999),样品平均回收率为99.12%,RSD为1.62%。3批黄柏经过炮制后其含有盐酸小檗碱的含量均降低。结论该方法简便、快速准确、灵敏度高,重现性好。有助于为黄柏合理的炮制工艺提供依据,可用于其炮制品的质量控制。     

HPLC-UV法同时测定不同产地黄柏中小檗碱和巴马汀的含量

目的:建立不同产地黄柏中有效成分小檗碱和巴马汀的含量测定方法.方法:采用高效液相色谱法,测定了十六个产地黄柏中小檗碱和巴马汀的含量.色谱柱:Diamonsil~(TM)C_(18)(250mm×4.6mm,5μm):检测波长:345nm;温度:20℃;流动相:水-乙腈-冰醋酸-三乙胺(67:33:0.3:0.8.v/v);流速:1mL·min~(-1).结果:建立了同时测定黄柏中小檗碱和巴马汀的高效液相色谱法,产自广西桂林的黄柏中小檗碱含量最高,产自那坡的黄柏中巴马汀含量最高.结论:本法简便、快速、准确,适用于黄柏中小檗碱和巴马汀的含量测定.     

黄柏与甘草及桑寄生共煎对小檗碱提取率影响的研究

目的 :研究黄柏与甘草及桑寄生共煎对小檗碱提取率的影响。方法 :采用高效液相色谱法对黄柏单煎和与甘草及桑寄生共煎液中小檗碱提取量进行测定 ;色谱柱为HypersilBDSC18,流动相为乙腈 -33mmol/L磷酸二氢钾 -三乙胺 (20∶72∶0. 1) ,检测波长为345nm ,柱温为25℃ ,流速为1. 0ml/min。结果 :黄柏与甘草及桑寄生共煎使小檗碱的提出率明显降低。结论 :对含有上述药物的制剂在制定提取工艺时 ,将黄柏单独煎煮提取较好。     

HPLC同时测定糖足泡剂中盐酸黄柏碱和盐酸小檗碱的含量

目的建立同时测定糖足泡剂中盐酸黄柏碱和盐酸小檗碱含量的HPLC方法。方法色谱柱为Gemini C18(4.6×250 mm,5μm);流动相:乙腈(A)-0.03 mol/L磷酸二氢钾(B)梯度洗脱;流速:0.8 ml/min;检测波长:282 nm;柱温:30℃。结果盐酸黄柏碱和盐酸小檗碱分别在0.77~15.40μg/ml(r=0.9999,n=6),3.51~70.20μg/ml(r=0.9999,n=6)范围内线性关系良好,平均回收率97.9%,RSD分别为1.8%和1.3%。结论建立的方法简便、准确可靠,可用于测定糖足泡剂的质量控制。     

高效液相色谱法同时测定二妙丸中盐酸小檗碱和黄柏碱的含量

目的:建立用高效液相色谱法同时测定二妙丸中盐酸小檗碱和黄柏碱含量的方法。方法:色谱柱为Venusil C18(250 mm×4.6 mm,5μm);流动相为乙睛-0.05 mol.L-1磷酸二氢钾-三乙胺(用磷酸调pH=4)系统梯度洗脱;流速:1 mL.min-1;检测双波长:盐酸小檗碱为265 nm,黄柏碱为285 nm。结果:盐酸小檗碱的平均回收率为100.55%,RSD为2.76%(n=6),线性范围为1.623～81.17μg.mL-1r,=1.000 0;黄柏碱的平均回收率为99.32%,RSD为0.67%(n=6),线性范围为4.930～49.30μg.mL-1r,=0.999 9。结论:该方法灵敏简便、准确可靠、重复性好,可用于二妙丸的质量控制。     

黄柏胶囊中盐酸小檗碱的含量测定

目的采用高效液相色谱法测定黄柏胶囊中盐酸小檗碱的含量。方法以十八烷基硅烷键合硅胶为固定相，以乙腈-磷酸二氢钾(0.05 mol·L^-1)(28∶72)为流动相，流速1.0 mL·min^-1，检测波长349 nm。结果盐酸小檗碱进样量在0.04～0.20 μg范围内与峰面积具有良好的线性关系，r＝0.9995，回收率为99.99%，RSD＝0.56%(n＝5)。结论该方法操作简便，精密度好，结果准确可靠，适用于黄柏胶囊中盐酸小檗碱含量的测定。     

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.