超高效液相色谱法测定垂盆草颗粒中3种黄酮成分的含量

目的 建立超高效液相色谱法(UPLC)同时测定垂盆草颗粒中槲皮素、山柰素与异鼠李素的含量.方法 色谱柱Thermo Scientific AcclaimTM RSLC 120 C18(2.1 mm×100 mm,2.2 μm),流动相为甲醇-0.4％磷酸(50∶50),柱温为30℃,流速为0.3 mL·min-1,检测波长为360 nm,进样量1μL.结果 槲皮素、山柰素与异鼠李素的线性范围分别为:50～1 000 μg,20～400 μg,20 ～400 μg,且与峰面积呈良好线性关系(r=0.999 9,0.999 9,0.999 6),精密度、稳定性、重复性试验的RSD均＜2％,平均加样回收率(n=9)分别为99.35％,99.38％,98.91％,RSD分别为1.04％、0.98％、1.01％.结论 该法高效、准确、灵敏,可作为测定垂盆草颗粒中槲皮素、山柰素与异鼠李素含量的有效方法.     

垂盆草颗粒中槲皮素、山奈素与异鼠李素的HPLC 测定方法研究

目的：建立垂盆草颗粒中槲皮素、山奈素与异鼠李素的 HPLC 测定方法,为制定该制剂质量标准中定量测定方法及限度提供依据。方法色谱柱：SunFire-ODS C18（4．6 mm ×150 mm,5μm）流动相：以甲醇—0．4％磷酸溶液（50∶50）;流速：1．0 mL ·min －1,检测波长：360 nm,柱温：25℃,进样量：10μL。结果槲皮素、山奈素和异鼠李素的线性范围分别为34．35～429．43、10．83～135．37、12．39～154．94μg,平均加样回收率（n ＝6）分别为99．2％、99．3％和99．1％,RSD 分别为0．26％、0．29％和0．77％。结论该方法简单、快速、结果准确,重现性好,为垂盆草颗粒中槲皮素、山奈素与异鼠李素的测定提供可靠的方法。     

垂盆草片质量标准研究

目的：建立垂盆草片的质量标准。方法：采用薄层色谱法,对垂盆草进行定性鉴别;采用HPLC法对槲皮素、山柰素及异鼠李素进行含量测定。结果：该薄层色谱法专属性强;槲皮素、山柰素及异鼠李素的线性范围分别为4.94~98.82μg·ml-1、4.94~98.82μg·ml-1、1.07~21.36μg·ml-1,r值均为0.999 9,平均回收率分别为103.1%、100.6%、100.1%（RSD分别为2.2%、2.0%、2.0%,n=6）。结论：该方法专属性强、准确、重复性好,可用作垂盆草片的质量控制方法。     

一测多评法测定垂盆草颗粒中槲皮素、山奈素与异鼠李素含量

目的建立一测多评方法,测定垂盆草颗粒中槲皮素、山奈素与异鼠李素含量为控制垂盆草颗粒的质量提供依据。方法采用岛津C18色谱柱(250 mm×4.6 mm,5μm);流动性为甲醇-水(含0.4%磷酸溶液)=50∶40;流速为1.0 m L/min;检测波长360 nm;进样量:10μL;以槲皮素为参考建立山奈素和异鼠李素间的相对校正因子,考察其重现性,比较计算值与测得值的差异。结果槲皮素、山奈素和异鼠李素分别在32.36~426.43μg、11.56~136.87μg、10.45~155.68μg呈良好的线性关系,回归方程分别为:Y=3625.8X-13658(R2=0.9998)、Y=2682.9X-10253(R2=0.9999)、Y=3012.2X-11223(R2=0.9999)。平均加样回收率分别为99.3%、99.6%、98.5%(RSD均<1.0%)。用一测多评法对10批垂盆草颗粒中槲皮素、山奈素与异鼠李素进行测定的结果与外标法实测值基本一致。结论该方法可靠,结果准确,可用于垂盆草颗粒的质量控制。     

HPLC法同时测定南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量

目的 建立南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量测定方法。方法 采用Diamond—C18色谱柱;甲醇-0．4％磷酸溶液（50：50）为流动相;流速1．0mL·min;检测波长360nm。结果 线性范围：槲皮素0．1184～0．8288μg,r=0．9995,山奈酚0．0258～0．1806／μg,r=0．9994,异鼠李素0．0444～0．3108μg,r=0．9994;平均回收率：槲皮素99．51％,RSD=2．10％（n=6）,山奈酚101．03％,RSD=2．08%（n=6）,异鼠李素99．80％,RSD=2．09％（n=6）。结论 该法操作简便、准确,重复性好,可用于南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量测定。     

HPLC法同时测定垂盆草片中槲皮素等3种成分

目的建立同时测定垂盆草片中槲皮素、山奈素、异鼠李素3种成分的HPLC方法。方法采用Imertsit ODS-SP色谱柱（150×4．6mm,5μm）,柱温35℃,流动相为甲醇旬．4％磷酸溶液（45：55）,流速1．0mL·min^-1,检测波长360nm．结果3种成分均达到基线分离,槲皮素、山奈素、异鼠李素3种成分分别在0．94μg·mL^-1～37．6μg·mL^-1,0．808μg·mL^-1-16．16μg·mL^-1,1．116μg·mL^-1～22．32μg·mL^-1。的浓度范围内与峰面积线性关系良好。平均回收率分别为101．95％、99．01％、101．00％。结论该方法准确可靠,可用于垂盆草片的质量控制。     

HPLC法同时测定土荆芥中槲皮素、山奈素、异鼠李素的含量

目的：建立土荆芥中槲皮素、山奈素、异鼠李素含量的HPLC测定方法。方法采用 DIKMA Platisil ODS C18色谱柱（250mm ×4．6mm,5μm）,流动相为乙腈－0．3％磷酸（38∶62）,流速为1．0mL? min －1,检测波长为370nm,柱温为30℃。结果槲皮素进样量在0．0559～3．3552μg与峰面积线性关系良好,r＝0．9999,平均回收率为104．82％,RSD为1．83％（n ＝6）;山奈素进样量在0．1112～6．6720μg与峰面积线性关系良好,r＝0．9999,平均回收率为100．82％,RSD为2．43％（n＝6）;异鼠李素进样量在0．0128～0．7680μg与峰面积线性关系良好,r＝0．9999,平均回收率为99．49％,RSD为2．55％（n＝6）。结论本法准确、重复性好,可用于同时测定土荆芥中槲皮素、山奈素、异鼠李素的含量。     

HPLC法测定通淋颗粒中槲皮素和山奈素的含量

目的 同时测定通淋颗粒中槲皮素和山奈素的含量.方法 采用HPLC法,样品经80%甲醇超声,酸水解后,采用色谱柱C18柱(4.6 mm×250 mm,5 μm),甲醇-0.4%磷酸(50∶50)为流动相,流速:1.0 ml·min-1,检测波长360 nm,柱温:30 ℃.结果 槲皮素在0.128～2.304 μg(r2=0.9996)和山奈素在0.1255～2.259 μg(r2=0.9999)范围内均呈良好的线性关系,槲皮素和山奈素的加权回收率分别为99.93%(RSD=1.06%)和99.23%(RSD=0.75%)(n=6).结论 本方法操作简便、重复性好、结果准确可靠,可满足通淋颗粒的质量控制要求.     

HPLC法测定柴胡属植物地上部分黄酮类含量

目的:建立柴胡属植物地上部分中槲皮素、异鼠李素、山奈素HPLC的含量测定方法。方法:伊利特-C18柱(250 mm×4．6 mm,5μm),以甲醇-水(60:40),磷酸调节pH至3．0为流动相,流速:1．0 ml·min-1,检测波长:360 nm。结果:槲皮素在0．70-5．57μg(r槲=0．999 7),异鼠李素在0．16～1．30μg(r异=0．999 2),山奈素在0．09-0．71μg(r山=0．999 4)范围内呈良好的线性关系。平均回收率分别为97．0％,96．8％,97．4％,RSD分别为2．2％,2．2％,1．1％。结论:本法专属性强,灵敏度高,重现性好,适于黄酮类成分的分离与测定。     

HPLC法同时测定宫炎平胶囊中没食子酸、槲皮素、山柰素的含量

目的:建立宫炎平胶囊中没食子酸、槲皮素和山奈素的含量测定方法。方法:应用高效液相色谱法测定,色谱柱为Agilent-SB C18(250 mm×4.6 mm,5μm),以甲醇和0.3%磷酸水溶液为流动相进行梯度洗脱,流速为1.0 ml·min-1,柱温30℃。结果:没食子酸、槲皮素和山奈素的线性范围分别为98.200~491.000μg·ml-1(r=0.999 9)、7.520~37.600μg·ml-1(r=0.999 9)、4.940~24.700μg·ml-1(r=0.999 9);平均加样回收率分别为96.74%(RSD=1.33%)、98.18%(RSD=1.70%)、97.04%(RSD=1.28%)结论:该方法简便易行、准确、重复性好。     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.