人血浆中西酞普兰的HPLC-荧光法测定及其药物动力学

建立了HPLC-荧光法测定人血浆中西酞普兰的浓度,并进行了药物动力学研究.以盐酸普萘洛尔为内标,采用Dikma柱,以乙腈-0.03mol/L醋酸铵缓冲液(33:67)为流动相,激发波长和发射波长分别为236和306nm.血浆中西酞普兰线性范围为1～75ng/ml,定量限为1ng/ml,方法回收率92.8%～98.8%.18名健康志愿者口服西酞普兰40mg后,血浆中的Cmax为(54.54±7.83)ng/ml,Tmax为(3.06±0.42)h,AUC0～132为(2351.1±389.9)ng·ml-1·h,AUC0～∞为(2636.5±444.9)ng·ml-1·h,t1/2为(40.45±8.68)h.     

高效液相色谱-荧光法测定人血浆中西酞普兰含量

目的:建立快速灵敏的高效液相色谱-荧光检测法测定西酞普兰的血药浓度。方法:血浆样品加入内标(右美沙芬)后用乙醚提取,乙醚层经空气流吹干后用流动相定容,以40μL 进样。色谱柱为 Kromasil KR100-5 C_(18)(250 mm×4.6 mm,5μm),流动相采用0.05 mol·L~(-1),pH=4.23的磷酸二氢钠缓冲液-乙腈(63:37),流速为0.6 mL·min~(-1),荧光激发波长为240 nm,发射波长300 nm,内标法定量。结果:本法的最低定量浓度为1.3 ng·mL~(-1),在1.3～260 ng·mL~(-1)浓度范围内线性良好,r=0.9999。日内精密度为1.1%～2.3%,日间精密度为1.7%～4.0%,萃取回收率为84.4%～88.3%,准确度为86.2%～95.6%,整个分析过程在15 min 内完成。结论:该法准确、精密度高、重现性好,适用于人血浆中西酞普兰的动力学研究和生物等效性研究。     

高效液相色谱法测定人血浆中西酞普兰浓度

目的:测定人血浆中西酞普兰的浓度.方法:日本HiQ sil C18V(4.6 mm×150 mm,5 μm)色谱柱,检测波长为240 nm,流动相为0.05 mol·L-1磷酸二氢钾缓冲液-乙腈(70:30)(pH4.58),流速:0.8 mL·min-1,以西沙比利为内标.结果:西酞普兰在16.9～508.0μg·L-1范围内呈良好线性关系(r=0.999 4),提取回收率在78.6%～86.7%之间,日内和日间RSD分别为0.9%～4.1%和2.1%～8.6%.结论:该法简便、快速、准确、重现性好,适用于临床西酞普兰的药物浓度监测.     

反相高效液相色谱荧光检测法测定血浆中西酞普兰浓度

目的:建立反相高效液相色谱法测定人血浆中西酞普兰浓度.方法:采用岛津公司LC-10Avp系列高效液相色谱仪,RF-10Axl荧光检测器,SIL-10Advp全自动进样器,Diamonsil C18(250mm×4.6mm,5μm ) 色谱柱,柱温:35℃.以CH3CN-0.05 mol·L-1 H3PO4 (36∶64)为流动相,普奈洛尔做内标,检测波长Ex=236nm,Em=306 nm.结果:标准曲线线性范围1.25～50.00 ng·mL-1;萃取回收率 84.96%～88.33%;方法回收率96.95%～105.2%;日内测定RSD为1.8%～6.1%,日间测定RSD为1.9%～5.3%.结论:本法快速、灵敏、准确、简便,可用于西酞普兰血药浓度测定和药动学及生物利用度研究.     

互为内标法测定患者血浆中氟西汀和西酞普兰的浓度

目的：建立测定人血浆中氟西汀、西酞普兰的高效液相色谱-荧光（HPLC-FLU）法,为临床用药提供参考。方法：氟西汀和西酞普兰互为内标,血浆样品碱化后经液-液萃取,以Ultimate XB-C18反相色谱柱进行分离,流动相为乙腈：0.02 mol·L-1磷酸氢二钠溶液（35∶65,pH 5.0）,流速为1.0 ml·min-1,柱温为30℃。荧光检测氟西汀和西酞普兰的激发波长为230nm,发射波长为305 nm。结果：氟西汀、西酞普兰血药浓度均在15.625～1 000μg.L-1范围内线性关系良好;平均回收率分别为100.72%和100.94%;分析方法的最低定量限均为5μg.L-1;日内、日间RSD均〈3%;冻融稳定性RSD均〈10%。结论：本法快速、简便、灵敏、准确,适用于氟西汀、西酞普兰的血药浓度监测及药动学研究。     

超高效液相串联质谱法测定大鼠西酞普兰及其代谢物的血药浓度

目的建立快速测定大鼠体内西酞普兰及其代谢物(N-氧化西酞普兰和N-去甲基西酞普兰)浓度的超高效液相串联质谱法。方法用乙酸乙酯萃取法处理血浆,色谱柱为ACQUITY UPLC BEH C18柱(50 mm×2.1 mm,1.7μm);流动相为乙腈-0.1%甲酸水,梯度洗脱,流速为0.4 m L·min-1;用正离子多离子反应监测(MRM)扫描,内标为曲马多。结果血浆中西酞普兰、N-去甲基西酞普兰和N-氧化西酞普兰的线性范围分别为0.1~10.0,0.1~10.0,0.01~2.00 ng·m L-1(r=0.999 1,0.999,0.999 6),定量下限分别为0.05,0.05,0.01ng·m L-1。其回收率分别在96.09%~105.33%。三者的日内、日间精密度均RSD<8.69%。结论该方法操作简便、快捷,灵敏度高,适于大鼠体内西酞普兰及其代谢物的药代动力学研究。     

反相高效液相色谱法测定人血浆中阿替洛尔的浓度

目的:建立反相高效液相色谱法测定血浆中阿替洛尔浓度.方法:以乙酸乙酯-异丙醇(8:1)为提取溶剂,液-液萃取法处理血浆样品.色谱柱:Phenomenex C18;流动相:0.05 mol·L-1磷酸二氢钠溶液(pH 3.2)-乙腈(90:10).检测波长:λex=230 nm,λem=310 nm.流速:1.0ml·min-1.结果:线性范围4.6～883.2 ng·ml-1;最低检测浓度为2.3 ng·ml-1;绝对回收率为95.1%;日内RSD小于3.9%,日间RSD小于5.5%.结论:方法灵敏准确,简便可行,适于阿替洛尔的治疗药物浓度监测.     

HPLC-荧光检测法测定人血浆中佐匹克隆的浓度

目的:建立测定人血浆中佐匹克隆浓度的方法。方法:采用液-液萃取法处理血浆后以高效液相色谱-荧光检测法进样测定,色谱柱为Kromasil C18,流动相为20mmol/L乙酸钠缓冲液(pH6.0)-乙腈-甲醇(67∶24∶9),流速为1.0ml/min,柱温为30℃,激发波长为307nm,发射波长为483nm。结果:佐匹克隆血药浓度在4～128ng/ml范围内线性关系良好(r=0.9980),定量下限为4ng/ml;方法回收率在96.3%～99.8%之间,提取回收率在75.4%～77.8%之间;日内、日间RSD均<10%。结论:本方法操作简便,专属性强、灵敏度高,可用于佐匹克隆的药动学研究。     

人血浆中文拉法辛浓度的HPLC测定及其药物动力学

建立了HPLC-荧光法测定文拉法辛的血药浓度.采用液液萃取处理血浆样品,色谱柱为C8柱,流动相为乙腈-磷酸盐缓冲液(30:70,pH 5.5),激发和发射波长分别为276和598nm,流速1.5ml/min.文拉法辛在5～2000ng/ml的浓度范围内线性关系良好.方法提取回收率为96.9%～104.3%,日内和日间RSD均小于8%.10名健康志愿者空腹单剂量口服150mg盐酸文拉法辛缓释胶囊,药物动力学参数Cmax、Tmax、AUC0→∞、t1/2分别为(168±12.3)ng/ml、(6.1±1.8)h、(4510±1462)ng·h·ml-1和(5.7±0.7)h.     

犬血浆中马来酸曲美布汀的HPLC法测定

建立了高效液相色谱法测定犬血浆中的马来酸曲美布汀.采用C18色谱柱,以0.05 mol/L磷酸二氢钾溶液(pH 3.5)-乙腈-甲醇(60∶35∶5)为流动相,检测波长267 nm.血浆中曲美布汀在20～1 000 ng/ml浓度范围内线性关系良好,日内、日间RSD小于15%,方法回收率为85%～115%.     

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.