二维高效液相色谱结合拦截技术测定舒巴坦血药浓度

目的建立二维高效液相色谱(2D-HPLC)结合中间柱拦截技术测定舒巴坦血药浓度的方法,并用于临床患者血液与透析液舒巴坦浓度的测定。方法第一维色谱柱为ASTON MVV C18柱(30mm×4.6 mm,5μm),流动相为甲醇-20 mmol·L~(-1)磷酸铵(5∶95,v/v)(p H=3.5),流速:0.8m L·min~(-1);第二维色谱柱为苯基乙基色谱柱(250 mm×4.6 mm,5μm),流动相为甲醇-20 mmol·L~(-1)磷酸铵(88∶12,v/v),流速:1.2 m L·min~(-1);中间柱为ASTON PLI中间柱(20 mm×4.6 mm,3μm),检测波长230 nm,温度:40℃,进样量:50μL。结果舒巴坦在2.55~249.20μg·m L~(-1)内与峰面积线性关系良好,r=0.9999,检测限为0.22μg·m L~(-1),绝对回收率为91.4%~102.1%,日间、日内精密度RSD均小于6%。结论本方法新颖,自动化程度高,样品前处理简单,检测结果准确稳定,适用于舒巴坦血药浓度及透析液的测定。     

高效液相色谱法测定鞭芍胶囊中芍药苷的含量

目的:建立测定鞭芍胶囊中芍药苷含量的高效液相色谱方法.方法:色谱柱为Hypersil ODS(200 mm× 4.6 mm,5 μm),流动相为乙腈-0.05% 磷酸溶液(14∶86),流速为 0.8 mL·min -1 ,检测波长为230 nm.结果:芍药苷进样量与色谱峰面积在 0.02 ～ 0.20 g·L -1 范围内呈良好的线性关系(r= 0.999 8 ),平均加样回收率为 100.9% ,RSD为 2.4% (n=6).结论:该法操作简单、精密度高、重现性好,是控制鞭芍胶囊内在质量的有效方法.     

HPLC测定头孢地尼胶囊的含量

目的建立测定头孢地尼胶囊含量的方法。方法采用HPLC法,色谱柱用C18柱(4.6mm×150mm,5μm);流动相为0.029mol·L-1磷酸氢二铵溶液(磷酸调pH5.0)-乙腈(86∶14);流速0.5ml·min-1;柱温25℃;检测波长286nm;进样量20μl。结果头孢地尼在10～100μg·ml-1范围内峰面积与浓度呈良好的线性关系(r=0.9999);平均回收率为100.19%,RSD=0.45%(n=3)。结论所用方法精密度、重复性和回收率良好,结果准确可靠。     

氨咖黄敏胶囊中胆红素的含量测定

目的采用反相高效液相色谱法测定氨咖黄敏胶囊中胆红素的含量.方法采用DiamonsilTMC18色谱柱(4.6 mm×200 mm,5μm),以甲醇-氯仿-1%磷酸溶液(85:13:2)为流动相,流速1.0 mL·min-1,检测波长450nm.结果胆红素在3.97～79.5μg·mL-1峰面积与浓度呈良好的线性关系(r=0.997),平均回收率为99.95%,RSD为0.38%(n=9).结论方法简便、精密度高、重现性好、结果准确可靠.     

HPLC法同时测定腰腿痛丸中麻黄碱、士的宁及马钱子碱的含量

目的采用HPLC法同时测定腰腿痛丸中麻黄碱、士的宁和马钱子碱的含量。方法采用HPLC法。色谱柱为安捷伦C18色谱柱(250mm×4.6mm,5μm);流动相为乙腈-1mL·L-1磷酸(12∶88);流速为1.0mL·min-1;检测波长为210和254nm;柱温为30℃。结果麻黄碱在1.0²5μg·mL-1范围内线性关系良好(r=0.999 9),平均回收率为98.7%;士的宁在2.025μg·mL-1范围内线性关系良好(r=0.999 9),平均回收率为98.7%;士的宁在2.0⁵0μg·mL-1范围内线性关系良好(r=0.999 8),平均回收率为99.1%;马钱子碱在250μg·mL-1范围内线性关系良好(r=0.999 8),平均回收率为99.1%;马钱子碱在2⁵0μg·mL-1范围内线性关系良好(r=0.999 9),平均回收率为97.9%。3批腰腿痛丸中麻黄碱、士的宁及马钱子碱的平均含量分别为93.1,106.7和76.8μg·g-1。结论该方法灵敏,精密度好,结果准确可靠。     

RP-HPLC法测定维胺酯乳膏中维胺酯的含量

目的:测定维胺酯乳膏中维胺酯的含量。方法:采用 RP-HPLC,Kromasil C_(18)(4.6mm×250mm,5μm)色谱柱,甲醇-水(93∶7)为流动相,检测波长371nm。结果:维胺酯在2.068-20.68μg·mL~(-1)范围内线性关系良好(r=0.9999),平均回收率98.72%(n=5),方法精密度 RSD=0.9%。结论:方法简便、快速,准确。     

HPLC法测定盐酸丁卡因滴眼液中羟苯乙酯的含量

目的采用高效液相色谱法测定盐酸丁卡因滴眼液中抑菌剂羟苯乙酯的含量。方法色谱柱为Agilent HC-C18色谱柱(250mm×4.6mm,5μm);流动相为0.005mol·L-1醋酸胺溶液(每1 000mL含三乙胺10mL;冰醋酸调pH5.0)-乙腈(55∶45);检测波长为256nm;流速1.0mL·min-1。结果羟苯乙酯质量浓度在14.31~143.12μg·mL-1范围内,与其峰面积呈良好的线性关系(r=1),平均回收率为99.0%(RSD=1.36%)。结论该方法专属性强、测定结果准确、精密度高、线性好,适用于检测盐酸丁卡因滴眼液中羟苯乙酯的含量。     

高效液相色谱法测定炉甘石薄荷脑洗剂中苯酚的含量

目的测定炉甘石薄荷脑洗荆中苯酚的含量.方法采用HPLC法以Kromasil C18色谱柱(4.6 mm x250mm,5μm),流动相为甲醇-水(6535),检测波长270 nm.结果苯酚的浓度在40～180 mg·L-1范围内线性关系良好(r=0.999 8),平均回收率为100.1%,方法精密度ILSD=0.45%( n=5).结论该法简便、快速、准确,可作为炉甘石薄荷脑洗剂中苯酚的含量测定方法.     

HPLC法监测人血清中茶碱类药物浓度

目的建立同时测定茶碱类药物血药浓度的高效液相色谱法。方法筛选得到的最佳色谱条件为:采用色谱柱Ultimate XB-C18柱(4.6×200mm,5μm),以咖啡因为内标,甲醇-乙腈-水(20∶3∶77)为流动相,流速1.0m L·min-1,检测波长273nm,柱温30℃。采用先沉淀后提取的方法进样测定。结果多索茶碱和茶碱分别在1.5~24mg·L-1(r=1.0000)和2.5~40mg·L-1(r=0.9996)范围内线性关系良好。多索茶碱的低、中、高浓度平均回收率为100.07±5.30%,日间精密度和日内精密度RSD均小于6.5%。茶碱的低、中、高浓度回收率为101.72±4.92%,日间精密度和日内精密度RSD均小于5.5%。结论该方法灵敏度高、准确、重复性好,适用于多索茶碱和茶碱血药浓度的测定。     

HPLC法测定发酵液中非达霉素的含量北大核心CSCD

目的:建立高效液相色谱法定量检测微生物发酵液中的非达霉素。方法:采用Agilent 5 TC-C18色谱柱(250 mm×4.6 mm,5μm),以甲醇-0.1%甲酸水为流动相,流速1.0 m L·min^(-1),检测波长228 nm,柱温40℃。结果:非达霉素质量浓度在6.59~79.08μg·m L^(-1)范围内与峰面积呈现良好的线性关系(r=0.999 9,n=7);平均回收率为98.6%(n=9,RSD=0.91%);精密度、重复性和稳定性良好,RSD均小于2%;3批发酵液中非达霉素的测定结果分别为59.4、52.5、53.6μg·m L^(-1)。结论:本方法可用于非达霉素的菌株筛选以及发酵过程产物的控制。     

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.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.