HPLC法测定非诺贝特缓释片中非诺贝特

目的采用HPLC法测定非诺贝特缓释片中非诺贝特。方法采用Dikma C18色谱柱(200 mm×4.6 mm,5μm);流动相:甲醇–水(90∶10);检测波长:288 nm;柱温:25℃;体积流量:1.0 mL/min;进样量10μL。结果非诺贝特在2.0~12.0μg/mL与其峰面积呈良好的线性关系(r=0.999 9);检测限和定量限分别为10、30 ng/mL;平均回收率为99.86%,RSD值为0.72%(n=9)。结论该方法准确、简便,可用于非诺贝特缓释片中非诺贝特的质量控制。     

HPLC法测定维格列汀原料药中对映异构体

目的建立测定维格列汀原料药中对映异构体的HPLC测定方法。方法采用Chirapak IC-3手性色谱柱(250 mm×4.6 mm,3μm);流动相:正己烷–乙醇–三乙胺(150∶750∶1);检测波长:210 nm;体积流量:0.6 mL/min;进样量:20μL;柱温:35℃。结果在上述色谱条件下,维格列汀与其对映异构体的分离度大于5;最低定量限为1.2μg/mL,最低检测限为0.3μg/mL;维格列汀和对映异构体在1.2～30.0μg/mL与峰面积呈良好的线性关系。对映异构体的加样回收率为99.67%,RSD值为2.1%;色谱参数变动对对映异构体的检测没有影响。结论本方法灵敏度高,重现性好,适用于维格列汀中对映异构体杂质的检查。     

顶空气相色谱法测定甲基橙皮苷原料中有机溶剂的残留量

目的建立甲基橙皮苷中有机溶剂乙醇和正丁醇的气相色谱测定方法。方法顶空气相色谱法,色谱条件:色谱柱为Agilent DB-624毛细管柱(30 m×0.32 mm,1.80μm);检测器:氢火焰离子化检测器,进样口温度220℃,检测器温度260℃;柱温:初始温度60℃,保温10 min,以40℃/min升至180℃,保温5 min;载气:氮气;流速6.00 mL/min;分流比10∶1。结果乙醇浓度在10.066～201.32μg/mL范围内,正丁醇浓度在5.032～100.64μg/mL范围内具有良好的线性范围;乙醇的平均回收率为98.6%(n=6),RSD为0.96%,正丁醇的平均回收率为99.5%(n=6),RSD为1.6%。结论本方法简便、快速、灵敏,是控制甲基橙皮苷残留有机溶剂的可靠测定方法。     

复方毛冬青注射液中甲基正壬酮的含量测定

目的:为复方毛冬青注射液建立专属性含量测定方法。方法:采用气相色谱法对甲基正壬酮进行含量测定。色谱柱:HP-1(Aglient 19091z-413,30 m×320μm×0.25μm);氮气为9.23 psi,流量1.8 mL/min;空气400 mL/min;氢气40 mL/min;柱温80℃(2 min)至220℃,升温速率为15℃/min;再升至270℃(13.75 min),升温速率为40℃/min;汽化室240℃;检测室280℃;进样量1μL;检测器为FID。结果:线性范围为1.0~20.0μg/mL,平均回收率为99.05%,RSD=1.29%(n=6)。结论:本法操作简单、可靠、具有实用性。     

气相色谱-电子捕获检测器法测定奥美沙坦酯原料药中基因毒性杂质碘甲烷

目的采用气相色谱–电子捕获检测器(GC-ECD)法测定奥美沙坦酯原料药中基因毒性杂质碘甲烷。方法采用Agilent DB-624毛细管色谱柱(30 m×0.32 mm×1.8μm),初始柱温40℃,保持5 min,以20℃/min升至200℃,保持2min,进样口温度160℃,检测器温度300℃,载气为N2,体积流量为1.5 mL/min,分流比10∶1,进样量1μL。结果碘甲烷在0.03～0.60μg/mL线性关系良好;检测限为2.0 ng/mL,定量限为6.0 ng/mL;平均回收率为98.0%,RSD值为1.8%。结论该方法专属性强、准确性高、灵敏度高、耐用性好,可用于奥美沙坦酯中基因毒性杂质碘甲烷的测定。     

HPLC法测定依维莫司原料药中依维莫司

目的建立HPLC法测定依维莫司原料药中依维莫司。方法采用Hypersil-ODS C18色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈–水(70∶30);检测波长:276 nm;柱温:50℃;体积流量:0.8 m L/min;进样量:10μL。结果依维莫司在63~1 008μg/m L线性关系良好(r=0.999 7),平均回收率为100.45%,RSD值为0.64%(n=9)。结论所建立的操作简便,专属性强,可用于依维莫司原料药的质量控制。     

苦参配方颗粒质量控制研究

目的建立苦参配方颗粒的质量控制标准。方法采用薄层色谱(TLC)法鉴别苦参配方颗粒中苦参碱、槐定碱和氧化苦参碱。采用高效液相色谱(HPLC)法测定苦参配方颗粒中苦参碱和氧化苦参碱,色谱条件为Lichrospher-5NH2色谱柱(250mm×4.6mm,5μm);流动相:乙腈–无水乙醇–3%磷酸溶液(80∶10∶10);体积流量:1.0mL/min;柱温:30℃;检测波长:220nm;进样量:10μL。结果苦参碱、槐定碱和氧化苦参碱在TLC色谱图中相应的位置上显相同的斑点。苦参碱在0.15～1.58μg、氧化苦参碱在0.17～1.68μg呈良好的线性关系,回收率分别为98.3%、101.5%,RSD值分别为1.35%、1.73%。结论该方法快速、灵敏、准确,可用于苦参配方颗粒的质量控制。     

大黄、枳实、厚朴不同炮制品对小承气汤中药效组分的影响

目的研究大黄、枳实、厚朴饮片变化对小承气汤药效组分的影响,以期为临床的合理应用和饮片的质量标准提供参考。方法采用HPLC法分别测定各类成分。大黄游离蒽醌类(芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚):Syncronis C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相:甲醇–0.1%磷酸;梯度洗脱;体积流量0.8 m L/min;柱温30℃;进样量10μL;检测波长254 nm。大黄结合蒽醌类(番泻苷B、番泻苷A):Syncronis C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈–0.05%磷酸;梯度洗脱;柱温30℃;进样量10μL;检测波长340 nm。枳实黄酮苷类(芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷):Syncronis C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈–水;梯度洗脱;体积流量0.7 m L/min;柱温40℃;进样量10μL;检测波长283 nm。厚朴木脂素类成分(和厚朴酚、厚朴酚):Syncronis C_(18)色谱柱(250 mm×4.6mm,5μm);流动相:乙腈–0.05%磷酸;梯度洗脱;体积流量0.7 m L/min;柱温30℃;进样量10μL;检测波长294 nm。结果小承气汤配伍剂量(大黄12 g–炒枳实9 g–姜厚朴6 g)不变,大黄、枳实、厚朴饮片改变时,小承气汤药效组分总量变化规律为:大黄–枳实–姜厚朴酒大黄–炒枳实–姜厚朴熟大黄–炒枳实–姜厚朴大黄炭–炒枳实–姜厚朴≈小承气汤大黄–炒枳实–厚朴,变化率分别为酒大黄组(6.561%)、熟大黄组(4.222%)、大黄炭组(0.118%)、枳实组(30.186%)、厚朴组(-11.218%)。除大黄炭组外,其余组的药效组分总量皆明显变化,其中枳实组变化最明显。结论同一味药材的不同炮制品在小承气汤处方中药效组分不同,对其他药味的影响亦不同,在小承气汤处方配伍中不可随意替代。     

HPLC法测定补气生血口服液中三种组分的含量

目的建立以高效液相色谱测定补气生血口服液中黄芪甲苷、二苯乙烯苷和阿魏酸含量的方法。方法三组分均采用phenomenex C_(18)(250 mm×4.6 mm,5μm)色谱柱;黄芪甲苷测定的流动相为水-乙腈(60:40);柱温:30℃;流速:1.0 mL/min;进样量:10μL;ELSD检测器;漂移管温度:105℃;气流速度:2.5 L/min。二苯乙烯苷与阿魏酸测定流动相为乙腈-水(20:80);流速1.0 mL/min;检测波长:248 nm;柱温30℃。结果补气生血口服液中黄芪甲苷在0.22~5.72μg/mL范围内线性关系良好,加样回收率为91.2%。在相同色谱条件下,二苯乙烯苷与阿魏酸分离度好,二苯乙烯苷在9.83~393.00μg/m L范围内、阿魏酸在1.015~50.756μg/mL范围内线性关系良好,加样回收率分别为90.7%和92.8%。结论该方法简便、快捷、专属性强,为其质量控制与含量测定提供了重要参考。     

HPLC法测定复方盐酸林可霉素滴鼻剂中盐酸林可霉素和利巴韦林

目的建立同时测定复方盐酸林可霉素滴鼻剂中盐酸林可霉素和利巴韦林的HPLC方法。方法采用高效液相色谱梯度洗脱法。Shim-pack VP-ODS C18色谱柱(150 mm×4.6 mm,5μm);流动相:甲醇–0.05 mol/L四硼酸钠溶液(用磷酸调p H值至6.0);梯度洗脱;柱温:30℃;体积流量:1 m L/min;检测波长:210 nm;进样量:10μL。结果盐酸林可霉素和利巴韦林分别在0.202 8~2.028 0μg和0.101 6~1.016 0μg与峰面积具有良好的线性关系,平均回收率分别为100.54%、99.95%,RSD值分别为1.11%、1.45%(n=9)。结论建立的方法简便、灵敏、准确,为复方林可霉素滴鼻剂的质量控制提供支持。     

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.

     

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.