HPLC法测定枳术颗粒中枳实的5种黄酮类物质

目的：建立枳术颗粒5种黄酮类物质含量控制方法。方法：采用高效液相色谱法在同一色谱条件下测定枳术颗粒中柚皮苷、橙皮苷、新橙皮苷、柚皮素、橙皮素含量。色谱条件：流动相乙腈-0.1%磷酸水梯度洗脱;检测波长为285 nm。结果：柚皮苷进样量在22.655～591.734 μg范围内呈良好线性关系,相关系数r²=0.9994;橙皮苷进样量在2.981～71.532 μg范围内呈良好线性关系,相关系数r²=0.9997;新橙皮苷进样量在22.152～531.648 μg范围内呈良好线性关系,相关系数r²=0.9994;柚皮素在3.015～72.378 μg范围内呈良好线性关系,相关系数r²=0.9997;橙皮素在2.868～68.85 μg范围内呈良好线性关系,相关系数r²=0.9990;5个成分的平均回收率分别为99.9%、100.4%、98.6%、99.9%和100.0%（RSD%＜3%）。结论：本方法准确可靠,可准确测定枳术颗粒中5种黄酮类物质的含量。     

HPLC同时测定衢枳壳中7种指标成分的含量

目的 建立HPLC同时测定衢枳壳中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、木犀草素、川陈皮素和桔皮素含量。方法 采用Agilent Extend C₁₈（4.6 mm×250 mm,5 μm）色谱柱,检测波长：330 nm,以乙腈-0.1%甲酸溶液为流动相,梯度洗脱,流速：1 ml·min^-1,柱温：30℃。结果 芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷、木犀草素、川陈皮素、桔皮素分别在23.30~1 164.80 ng,170.84~8 541.91 ng,17.22~861.20 ng,156.17~7 808.64 ng,3.13~156.48 ng,0.90~45.23 ng,0.85~42.27 ng内线性关系良好（r ≥ 0.999 5,n=6）,平均回收率分别为98.8%,101.3%,98.3%,96.8%,101.8%,101.7%,108.9%。结论 首次建立了衢枳壳中7个成分HPLC含量测定方法,该方法简便,结果准确,可为综合评价衢枳壳质量提供依据。     

HPLC双波长法同时测定炎热清片中6种有效成分的含量

目的 建立HPLC双波长法同时测定炎热清片中龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素和汉黄芩素6种成分的含量。方法 采用Agilent TC-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,流动相为甲醇∶0.2%磷酸水溶液（梯度洗脱）,柱温为35℃,流速为1.0 mL·min^-1,检测波长0~20 min为254 nm,同时检测龙胆苦苷、栀子苷、芒果苷,20~50 min为280 nm,同时检测黄芩苷、黄芩素、汉黄芩素。结果 龙胆苦苷、栀子苷、芒果苷、黄芩苷、黄芩素、汉黄芩素分别在18.92~189.2 ng（r=0.999 9）,107.44~1 074.4 ng（r=0.999 9）,9.82~98.2 ng（r=0.999 9）,767.68~7 676.8 ng（r=1.000 0）,100.94~1 009.4 ng（r=0.999 9）,49.84~498.4 ng（r=0.999 9）线性良好,平均回收率（n=9）分别为98.31%,98.35%,98.40%,98.03%,98.46%,98.24%,RSD分别为0.4%,0.3%,0.3%,0.4%,0.4%,0.3%。结论 该方法灵敏、简便、准确,可用于炎热清片的质量控制。     

HPLC法同时测定胃痛丸中柚皮苷、新橙皮苷的含量

摘 要 目的：建立胃痛丸中柚皮苷、新橙皮苷的高效液相色谱法（HPLC）含量测定方法。方法: 采用HPLC,色谱柱为Agilent TC C18 柱 ,以乙腈-水（20∶80）为流动相,流速为1.0ml·min^-1,检测波长为283 nm,柱温为30℃。 结果: 柚皮苷和新橙皮苷分别在0.027～4.552 μg( r=0.999 9) 、0.029～4.016 μg( r=0.999 8) ,进样量与峰面积呈良好的线性关系,平均加样回收率分别为102.19%、103.60%,RSD分别为2.88% 、1.79%( n=9)。 结论:方法简便、准确、灵敏、重复性好,可作为胃痛丸中的柚皮苷和新橙皮苷含量控制方法。     

HPLC同时测定半夏糖浆中琥珀酸、橙皮苷、甘草苷3种成分的含量

目的 建立HPLC同时测定半夏糖浆中琥珀酸、橙皮苷、甘草苷3种成分的含量。方法 采用C₁₈色谱柱（4.6 mm×150 mm,5μm）,以乙腈-0.2%磷酸溶液（18：82）为流动相,流速1.0 mL·min^-1,检测波长207 nm,柱温为常温。结果 3种待测成分分离度良好,阴性无干扰;3个成分线性范围分别为4.727~118.17 μg·mL^-1（r=0.999 9）,2.474~61.857 μg·mL^-1（r=0.999 6）,2.469~61.725 μg·mL^-1（r=0.999 9）;琥珀酸、橙皮苷、甘草苷平均回收率（n=9）分别为100.8%,99.3%,100.2%,RSD分别为1.3%,1.2%,1.8%,3批中琥珀酸、橙皮苷、甘草苷含量范围分别为0.072 2~0.079 4、0.029 9~0.034 8,0.022 8~0.029 0 mg·mL^-1。结论 该方法操作简单,缩短了分析时间,重复性好,可为半夏糖浆质量控制提供参考。     

六味开胃消食颗粒质量标准的建立

摘 要 目的： 建立六味开胃消食颗粒的质量标准。方法: 采用TLC法对六味开胃消食颗粒中的枳壳、木香、白术和山楂等药材进行了薄层色谱鉴别;采用HPLC法测定君药枳壳药材中柚皮苷与新橙皮苷的含量,色谱柱为Sinochrom ODS BP（250 mm×4.6 mm,5 μm）,流动相：乙腈-0.1%磷酸（21∶79）;流速：1.0 ml·min^-1;检测波长：283 nm;柱温：30℃。结果: 薄层色谱法可鉴别出该制剂中的枳壳、木香、白术和山楂。柚皮苷在89.600～1.792×10³ μg范围内有良好的线性关系（r=0.999 8）,新橙皮苷在1.060×10²～2.120×10³ μg范围内有良好的线性关系（r=0.999 9）,柚皮苷平均回收率为98.67%,RSD为0.57%（n=6）,新橙皮苷平均回收率为98.07%,RSD为1.19%（n=6）。结论: 该方法准确、灵敏、重复性好,能有效的控制六味开胃消食颗粒的质量。     

HPLC同时测定复方鱼腥草片中4个有效成分的含量

目的 建立HPLC同时测定复方鱼腥草片中绿原酸、连翘酯苷A、槲皮苷和黄芩苷含量的方法。方法 采用CAPCELL PAK MG（4.6 mm×250 mm,5μm）色谱柱,流动相为乙腈（A）-0.1%磷酸溶液（B）,梯度洗脱（0~12 min,9% A;12~15 min,9%→15% A;15~32 min,15% A;32~48 min,15%→29% A;48~60 min,29% A）,流速1.0 mL·min^-1,柱温35℃,检测波长：330 nm。结果 绿原酸、槲皮苷浓度在2~100 μg·mL^-1内,连翘酯苷A浓度在4~200 μg·mL^-1内,黄芩苷浓度在12~600 μg·mL^-1内,与峰面积均呈良好的线性关系（r=0.999 9）;平均回收率（n=6）分别为98.0%,99.8%,99.1%和98.3%,RSD均<2.0%。结论 建立的HPLC可实现同时测定绿原酸、连翘酯苷A、槲皮苷和黄芩苷4种有效成分,方法快速、简便、结果准确,可为复方鱼腥草片提供更全面、可靠的质量控制方法。     

反相高效液相色谱法测定枳实中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷的含量

目的 建立反相高效液相色谱法同时测定枳实中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷的含量。方法 采用Promosil C18（250 mm×4.6 mm,5μm）色谱柱,流动相为乙腈-水（22：78）,流速为1.0 mL· min^-1,检测波长283 nm,柱温为25℃。结果 枳实中芸香柚皮苷、柚皮苷、橙皮苷、新橙皮苷分别在0.4116～2.470μg（r=0.9997）、0.413 2～2.066μg（r=1.000 0）、0.474 4～3.795μg（r=0.999 9）、0.198 2～1.982μg（r=0.999 7）与峰面积呈良好的线性关系,平均加样回收率分别为100.4%、103.3%、101.2%、99.2%;RSD分别为2.2%、1.6%、2.7%、2.0%。结论 该方法简便、准确、可靠,为枳实的质量控制提供一定的依据。     

RP-HPLC法测定前列癃闭通片中柚皮苷、新橙皮苷的含量

摘 要 目的: 建立反相高效液相色谱测定前列癃闭通片中柚皮苷、新橙皮苷的含量测定方法。方法: 色谱柱为WEL TECH Hisep C₁₈-T（250 mm×4.6 mm,5 μm）;流动相：乙腈-水（20∶80）（用磷酸调节pH值至3）;流速：1.0 ml·min^-1;检测波长：283 nm;柱温：25℃,进样量：10 μl。结果: 柚皮苷进样量在0.143～3.570 μg内呈良好的线性关系;新橙皮苷进样量在0.164～4.110 μg内呈良好的线性关系。两者的平均回收率分别为99.06%、98.40%,RSD分别为1.64%、1.29%（n=6）。结论: 该检测方法准确、可靠、重现性好,可用于前列癃闭通片中柚皮苷、新橙皮苷的含量测定。     

柴胡疏肝散水提取物的肠吸收特性研究

目的 建立同时测定芍药苷、阿魏酸、芸香柚皮苷、柚皮苷、新橙皮苷、甘草酸含量的超高效液相色谱（UPLC）法,研究柴胡疏肝散水提物的肠吸收特性。方法 采用大鼠在体单向肠灌流和外翻肠囊实验模型,应用UPLC法测定柴胡疏肝散水提物中指标性成分（芍药苷、阿魏酸、芸香柚皮苷、柚皮苷、新橙皮苷和甘草酸）在不同时间点、不同部位肠道吸收量,计算吸收动力学参数,考察其肠吸收特征。结果 在体单向肠灌流模型结果表明,芍药苷、芸香柚皮苷、新橙皮苷、柚皮苷和甘草酸均为中等程度吸收的化学成分,而阿魏酸为完全吸收;外翻肠囊模型结果表明,芍药苷在空肠部位,阿魏酸、柚皮苷和新橙皮苷在十二指肠部位,芸香柚皮苷在十二指肠和空肠部位的吸收最佳,而甘草酸在各肠段的吸收无显著性差异。结论 肠道对柴胡疏肝散中6种指标性成分均有吸收,阿魏酸较其他5种指标性成分更易透过肠壁进入血液循环;不同肠段对6种指标性成分的吸收具有选择性。     

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.

---

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.