高效液相色谱离子阱质谱联用法同时测定人血浆中曲马多和对乙酰氨基酚浓度

目的:建立快速、灵敏的液相色谱-电喷雾离子阱质谱法测定人血浆中曲马多和对乙酰氨基酚的浓度。方法:血浆样品经乙腈直接沉淀蛋白后,以乙腈-10 mmoL·L-1甲酸胺-0．1％甲酸(65:15:20)为流动相,采用Hypurity C18柱(150mm× 2·1 mm,5μm)分离,流速为0．2 mL·min-1,通过电喷雾离子化离子阱质谱,以二级质谱选择离子反应监测(SRM)方式进行检测。结果:曲马多和乙酰氨基酚的线性范围分别为5～1000及20-12800 ng·mL-1,最低检测浓度分别为1和10 ng· mL-1,平均相对回收率均为103．7％,日内和日间变异均<20％,每个样品测试时间仅为7．0 min。结论:本法简单、快速、灵敏、重现性好,是一种有效的检测人血浆中曲马多和对乙酰氨基酚浓度的方法。     

HPLC-MS／ESI测定人血清中丙戊酸浓度

目的:建立HPLC-MS／ESI测定人血清中丙戊酸浓度的方法,并用于临床血药浓度监测。方法:采用Waters XTerraTM MS C18色谱柱(150 mm×3．9 mm,5μm),以10 mmol·L-1醋酸铵水溶液-乙腈(60:40)为流动相,流速0．80 mL·min-1,柱温45℃,以双氯芬酸为内标。采用电喷雾电离源(ESI)负源,用各物质准分子离子峰的选择离子监测(SIR)进行定量分析,血清样品用乙腈直接沉淀后进样。结果:丙戊酸在7．25-188．10μg·mL-1浓度范围内线性关系良好(r=0．9997),最低检测浓度为0．50μg·mL-1(S／N≥3)。日内与日间RSD均小于10％(n=5),方法回收率在98．7％-105．1％之间。结论:该方法操作简单,结果准确,血药浓度监测时间很短,灵敏度高;适用于丙戊酸血药浓度监测及药代动力学研究。     

高效液相色谱法测定红毛五加中绿原酸的含量

目的:采用LC／MS2方法鉴定出中药红毛五加中绿原酸成分,并用高效液相色谱法测定其含量。方法:采用Zorbax SB-C18色谱柱(150 mm×4．6mm,5μm);流动相为乙腈和水(含0．16％甲酸)二元梯度洗脱系统;流速为1．0mL·min-1,检测波长327 nm。质谱条件:Agilent多级离子阱质谱仪:电喷雾化学电离源(ESI);负离子检出模式。结果:绿原酸的线性范围为 10-100 mg·L-1(r=0.9997),加样回收率为99．2％-101．8％;红毛五加中绿原酸的含量为0．58-5．45mg．g-1。结论:本方法灵敏、快速、重现性好,有助于红毛五加质量控制方法的研究。     

SPE-HPLC法测定复方甘草口服溶液中愈创甘油醚的含量

目的:建立测定复方甘草口服溶液中愈创甘油醚含量的方法。方法:选用Accu BounDⅡC18固相萃取小柱对样品进行纯化,采用Hypersil ODS(4．6 mm×250 mm,5μm)色谱柱,以0．05 mol·L-1磷酸二氢钾溶液-0．0025 mol·L-1庚烷磺酸钠水溶液-乙腈(20:20:4)为流动相,流速为1．0 ml·min-1,检测波长为220 nm。结果:线性范围为5-40μg·ml-1,r=0．999 9;回收率为101．0％,RSD为1．5％。结论:该法简便快速、结果准确可靠、可用于复方甘草口服溶液中愈创甘油醚的含量测定。     

RP-HPLC法同时测定壳聚糖止血海绵中3组分的含量

目的:建立壳聚糖止血海绵中地塞米松磷酸钠、替硝唑及克林霉素的RP-HPLC含量测定方法。方法:色谱柱为Dis- covery C18柱(150 mm×4．6 mm,5μm);流动相为乙腈-0．05 mol·L-1磷酸二氢钾缓冲液-三乙胺(35:65:0．3),用磷酸调pH至5．3;流速1．0 ml·min-1;检测波长210 nm。结果:地塞米松磷酸钠、替硝唑、克林霉素分别在2-10μg·ml-1(r=0．999 7)、6 -30μg·ml-1(r=0．999 8)、30-150μg·ml-1(r=0．999 8)线性范围内呈现良好的线性关系。3组分的平均回收率分别为99．4％(RSD=2．1％);98．7％(RSD=1．7％)、99．2％(RSD=1．9％)。结论:本法操作简便、快速、结果准确。可作为该制剂的质控方法。     

HPLC法测定洁阴舒洗剂中盐酸小檗碱的含量

目的:采用高效液相色谱法测定洁阴舒洗剂中盐酸小檗碱的含量。方法:用Hypersil C18(4．6 mm×250 mm,5 μm) 色谱柱,流动相乙腈-0．033 mol·L-1磷酸二氢钾(40:60,含0．5％三乙胺,磷酸调PH 3．1),流速1．0 ml·min-1;检测波长345 nm,柱温25℃。结果:盐酸小檗碱进样量在0．17-0．85μg之间,与峰面积呈良好的线性关系,r=1．000 0,平均加样回收率为 97．6％,RSD=0．4％(n=5)。结论:方法操作简单,快速。可作为测定本制剂中盐酸小檗碱的含量。     

反相高效液相法测定苦参素脂质体药物含量及包封率

目的:建立苦参素脂质体中药物含量及包封率测定的反相高效液相(RP-HPLC)法。方法:采用Di-monsil~(TM)ODS柱(200mm×4.6mm,5μm);流动相为乙腈-0.1％N_3PO_4水溶液(用三乙胺调pH3.13)(5:95);柱温:室温;流速:1mL·min~(-1);紫外检测波长:215nm。采用Sephadex G-50分离苦参素脂质体中的游离药物。结果:在本色谱条件下苦参素与辅料及溶剂峰分离良好,苦参素在20～100μg·mL~(-1)范围内线性关系良好(r=0.9999,n=5),回收率在99.90％～102.0％之间,日内RSD及日间RSD均小于2％(n=5)。结论:该方法准确可靠、简单快速,可用于苦参素脂质体含量及包封率的测定。     

HPLC法测定枸橼酸莫沙比利片的含量

目的:建立测定枸橼酸莫沙比利含量的HPLC法。方法:采用ODS色谱柱(250 ×4．6 mm,5 μm),磷酸二氢钾、庚烷磺酸钠溶液-乙腈(50:50)为流动相,流速1．2 ml·min-1检测波长为274 nm。结果:枸橼酸莫沙比利在5～50μg·ml-1范围内有良好线性关系(r=0．999 8),平均回收率为98．8％,RSD 1．3％。结论:本法,快速、准确     

HPLC法测定复方陈香胃片中橙皮苷的含量

目的:建立HPLC法测定复方陈香胃片中橙皮苷的含量。方法:用phenomenex C18柱(250mm×d4．6f mm,5μm),流动相为乙腈-0．02％磷酸溶液(25:75),流速1．0 ml·min-1在283 nm的波长处检测。结果:橙皮苷线性范围为0．43-2．16μg,r=0．999 9(n=5)。回收率为99．1％,RSD为1．5％(n=6)。结论:方法简便快速,准确,适合复方陈香胃片的质量控制。     

RP-HPLC 法同时测定成人血清中苦参素和阿德福韦酯的浓度方法学研究

目的：建立一种简易的测定联合服用苦参素和阿德福韦酯成人血清中苦参素和阿德福韦酯含量方法。方法采用Shim-pack VP-ODS C18色谱柱（250 mm ×4．6 mm,5μm）,流动相为甲醇∶水（70∶30）,流速为1．1 mL·min －1,检测波长为229 nm。结果苦参素保留时间4．6 min 左右,标准曲线的线性范围为0．0125～2 mg·L －1,r ＝0．9992。阿德福韦酯保留时间7．3 min 左右,标准曲线的线性范围为0．0125～0．75 mg·L －1,r ＝0．9991,苦参素和阿德福韦酯的日间、日内变异系数均在7％以下,回收率＞92％。结论该法简便,灵敏,快速,可用于苦参素和阿德福韦酯临床联用治疗的成人血药浓度的监测。     

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.