知柏地黄胶囊质量标准研究

目的:提高知柏地黄胶囊的质量标准。方法:采用TLC法对处方中山茱萸和熟地黄进行定性鉴别;采用HPLC法同时测定山茱萸中莫诺苷、马钱苷及牡丹皮中丹皮酚的含量。结果:TLC定性鉴别重复性好,阴性无干扰;莫诺苷、马钱苷和丹皮酚的线性范围分别为0.05~1.11μg(r=0.999 9)、0.07~1.52μg(r=0.999 9)、0.09~1.96μg(r=0.999 9);平均回收率(n=6)分别为101.6%、102.5%、103.4%,其RSD分别为2.2%、1.4%、1.1%。结论:该方法操作简便、准确,重复性好,可用于知柏地黄胶囊的质量控制。     

杞菊地黄口服液的质量标准提高研究

目的:为完善和提高杞菊地黄口服液的质量标准提供参考。方法:根据2015年版《中国药典》(四部)0502法对杞菊地黄口服液中枸杞子、菊花、牡丹皮进行薄层色谱(TLC)鉴别,分别以枸杞子、菊花和丹皮酚为对照,展开系统分别为三氯甲烷-乙酸乙酯-甲酸(6∶1∶0.5,V/V/V)、三氯甲烷-异丙醇-甲酸(10∶1∶0.5,V/V/V)和环己烷-乙酸乙酯(3∶1,V/V);采用高效液相色谱法(HPLC)同时测定杞菊地黄口服液中莫诺苷、马钱苷及丹皮酚的含量[色谱柱为InertSustain C_(18),流动相为乙腈-0.03%磷酸溶液(梯度洗脱),流速为1.0 mL/min,检测波长分别为240 nm(莫诺苷和马钱苷)和274 nm(丹皮酚),柱温为40℃,进样量为10μL]。结果:在枸杞子、菊花、牡丹皮TLC供试品图谱中,与对照药材/对照品图谱相应的位置上显相同颜色的斑点,且阴性对照无干扰。莫诺苷、马钱苷及丹皮酚检测质量浓度的线性范围分别为2.12～106.17、1.91～95.63、4.78～239.16μg/mL(R~2=0.999 9、0.999 9、0.999 8);定量限分别为2.12、1.91、2.39μg/mL;检测限分别为0.53、0.48、0.59μg/mL;精密度、重复性、稳定性试验的RSD均小于2%(n=6);平均回收率分别为98.27%、97.06%、97.65%,RSD分别为0.80%、1.18%、1.36%(n=6);耐用性试验的RSD均小于2%(n=3)。结论:本研究建立的方法操作简便、专属性强、耐用性好,可为完善杞菊地黄口服液的质量标准提供一定参考。     

HPLC法测定杞菊地黄丸中莫诺苷、马钱苷和丹皮酚的含量

目的:建立一种快速、准确和实用的HPLC方法,用于同时测定杞菊地黄丸中莫诺苷、马钱苷和丹皮酚的含量。方法:采用Phenomenex Gemini C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.3%磷酸水溶液为流动相梯度洗脱,流速1.0 m L·min-1,柱温40℃,检测波长为240 nm(莫诺苷、马钱苷)和274 nm(丹皮酚)。结果:莫诺苷、马钱苷、丹皮酚质量浓度分别在2.242~44.84μg·m L-1(r=0.999 9)、2.130~42.60μg·m L-1(r=1.000)、5.068~101.4μg·m L-1(r=1.000)范围内,与峰面积线性关系良好;平均回收率分别为96.2%(RSD=0.44%)、96.0%(RSD=0.21%)、103.0%(RSD=0.65%)。结论:本方法可作为杞菊地黄丸中莫诺苷、马钱苷和丹皮酚含量同时测定的方法,适用于杞菊地黄丸的质量控制。     

HPLC波长切换技术同时测定知柏地黄丸(浓缩丸)中莫诺苷、芒果苷、马钱苷和丹皮酚的含量

目的:建立HPLC法同时测定知柏地黄丸(浓缩丸)中莫诺苷、芒果苷、马钱苷、丹皮酚的含量。方法:采用Phenomenex C18色谱柱(4.6 mm×250 mm,5μm),以乙腈(A)-0.3%磷酸溶液(B)为流动相,梯度洗脱(0~5 min,5%A→7%A;5~26min,7%A;26~45 min,7%A→20%A;45~55 min,20%A→60%A;55~65 min,60%A),流速1 m L·min-1,柱温40℃,波长切换(0~45 min,在240 nm波长下检测莫诺苷、芒果苷和马钱苷;46~65 min,在274 nm波长下检测丹皮酚)。结果:莫诺苷、芒果苷、马钱苷和丹皮酚的线性范围分别为0.055 5~1.11μg(r=0.999 9)、0.040 2~0.804μg(r=0.999 9)、0.076 4~1.528μg(r=0.999 9)、0.098 2~1.964μg(r=0.999 9);平均回收率(n=6)分别为103.9%、102.0%、101.5%、102.2%,RSD分别为1.3%、1.3%、1.6%、0.82%。含量测定结果分别为1.117~1.501、0.375~0.447、1.083~1.562、2.306~2.857 mg·g-1。结论:该方法操作稳定准确,重复性好,可用于知柏地黄丸(浓缩丸)的质量控制。     

HPLC法同时测定知柏地黄丸中3种主要成分的含量

目的:建立同时测定知柏地黄丸中没食子酸、马钱苷、丹皮酚含量的方法。方法:采用高效液相色谱法。色谱柱为Zorb-ax C18柱,流动相为甲醇(A)-0.2%甲酸水溶液(B)梯度洗脱,流速为1.0mL.min-1,检测波长为252nm,柱温为30℃,进样量为10μL。结果:没食子酸、马钱苷、丹皮酚进样量分别在0.36～5.78(r=0.9998)、0.67～5.39(r=0.9999)、0.60～4.80(r=0.9997)μg范围内与峰面积积分值呈良好的线性关系;平均加样回收率分别为99.67%、99.16%、100.01%,RSD分别为1.26%、1.95%、1.78%(n=9)。结论:该方法准确、灵敏、可靠、重复性好,可用于知柏地黄丸的质量控制。     

六味地黄片质量标准研究

目的:建立六味地黄片的质量标准。方法:用薄层色谱(TLC)法对方中熟地黄、泽泻进行定性鉴别;用高效液相色谱法测定山茱萸中马钱苷的含量及牡丹皮中丹皮酚的含量。结果:熟地黄、泽泻的TLC斑点清晰,分离度好。马钱苷、丹皮酚的检测浓度分别在5.77～115.40μg.mL-1(r=0.9999)、5.12～102.50μg.mL-1(r=0.9998)范围内与各自峰面积积分值呈良好的线性关系;平均回收率分别为98.4%、97.08%,RSD分别为0.75%、1.07%(n均=6)。结论:所建标准可用于六味地黄片的质量控制。     

HPLC-PDA 波长切换法同时测定六味地黄膏中5种活性成分的含量

目的建立HPLC-PDA波长切换法同时测定六味地黄膏中莫诺苷、马钱苷、芍药苷、毛蕊花糖苷和丹皮酚的含量。方法采用Agilent HC-C_(18 )色谱柱(250 mm×4.6 mm,5μm);以乙腈(A)-2 mL·L~(-1)磷酸溶液(B)为流动相,梯度洗脱;流速:1.0 mL·min~(-1);莫诺苷、马钱苷、芍药苷、毛蕊花糖苷和丹皮酚的检测波长分别为240(0～33 min),240(0～33 min),230(33～37 min),334 (37～44 min)和274 nm(44～55 min);进样量:10μL;柱温:40℃。结果各指标成分分离度良好,莫诺苷、马钱苷、芍药苷、毛蕊花糖苷和丹皮酚5种成分的进样量分别在0.040 12～0.802 40(r_1=0.999 9),0.041 66～0.833 20(r_2=0.999 8),0.090 82～1.816 40(r_3=0.999 9),0.010 86～0.217 20(r_4=0.999 9)和0.092 94～1.858 80(r_5=0.999 9)μg范围内线性关系良好;平均回收率(n=8)分别为98.22%(RSD=0.96%),99.00%(RSD=0.87%),98.62%(RSD=0.77%),98.06%(RSD=0.91%)和99.18%(RSD=0.72%)。结论该方法操作简便、快速,结果准确、可靠,重复性好,可用于六味地黄膏的质量控制。     

除湿丸的质量标准研究

目的:为建立除湿丸的质量标准提供参考。方法:采用显微鉴别法、薄层色谱(TLC)法对方中牡丹皮、白鲜皮、当归、茜草、栀子进行定性鉴别;采用高效液相色谱法测定丹皮酚、黄芩苷的含量。色谱柱为Kromasil 100-5 C18,流动相为甲醇-水-磷酸(47∶53∶0.2,V/V/V),流速为1.0 ml/min,检测波长为280 nm,柱温为25℃,进样量为10μl。结果:显微鉴别中,白鲜皮、牡丹皮的显微特征明显,其他药材无干扰;TLC鉴别中,当归、茜草、栀子的特征斑点清晰,阴性对照无干扰。丹皮酚、黄芩苷进样量分别在0.106 24~2.124 8、0.059 04~1.180 8μg范围内与峰面积呈良好的线性关系(r=0.999 9、0.999 9);精密度、稳定性、重复性试验的RSD均≤2.06%;平均加样回收率分别为101.56%、100.16%,RSD分别为1.68%、1.13%(n=9)。结论:该方法操作简便、结果准确、重现性好,可作为除湿丸的质量控制标准。     

高效液相色谱法同时测定归芍地黄丸中4种化学成分的含量

目的建立高效液相色谱法同时测定归芍地黄丸中莫诺苷、马钱苷、芍药苷和丹皮酚的含量。方法色谱柱为Waters Symmetry C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.3%磷酸溶液,梯度洗脱,柱温为35℃,流速为0.8 mL·min-1,检测波长为240 nm,外标法计算含量。结果莫诺苷进样量在0.010 48~2.62μg与峰面积线性关系良好(r=0.9999),平均加样回收率为102.8%,RSD为1.4%;马钱苷进样量在0.00868~2.17μg与峰面积线性关系良好(r=0.9999),平均加样回收率为103.6%,RSD为1.1%;芍药苷进样量在0.007 696~1.15μg与峰面积线性关系良好(r=0.9943),平均加样回收率为102.3%,RSD为2.1%;丹皮酚进样量在0.018~2.25μg与峰面积线性关系良好(r=0.9998),平均加样回收率为102.2%,RSD为1.6%。结论该方法准确、简便、可行、重复性好,可有效控制归芍地黄丸的质量。     

HPLC法测定山茱萸配方颗粒中马钱苷的含量

目的:建立山茱萸配方颗粒中马钱苷的含量测定方法.方法:色谱柱为HangBang C18(200*4.6mm5μm):流动相为乙腈:水(15:85);检测波长240nm;流速1.0ml/min;柱温30℃.结果:马钱苷的检测浓度线性范围为0.005～0.025μg(r=0.9998),RSD=2.16%(n=5);平均加样回收率为98.79%,RSD=1.77%(n=6).结论:本法简便、快捷、准确,可用于山茱萸配方颗粒的含量测定.     

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.