高效液相色谱法测定奥硝唑的血药浓度及其人体药动学研究

目的:建立奥硝唑血浆浓度测定的反相高效液相色谱法,并用此法研究奥硝唑人体药动学.方法:采用Hypersil ODS色谱柱(250mm×4.6mm,10μm);以甲醇-0.01mol·L-1磷酸二氢钠(40∶60)为流动相,流速为1mL·min-1,检测波长318nm,甲硝唑作内标.结果:奥硝唑在0.5～32.0μg·mL-1浓度范围内呈线性,r=0.998 8.最低定量浓度0.5μg·mL-1.低、中、高浓度(1.0,8.0,32.0μg·mL-1)的方法回收率分别为86.2%,97.2%,96.8%;萃取回收率分别为73.8%,90.8%,92.9%,日内RSD<8%,日间RSD<11%.药动学研究表示,口服奥硝唑胶囊与片剂的药-时曲线均符合一级吸收、一级消除的一房室模型.结论:该法准确可靠,操作简便,适用于临床药动学研究及血药浓度监测.     

HPLC法同时测定丹香冠心注射液中3个成分的含量

目的:建立HPLC法同时测定丹香冠心注射液中丹参素钠、原儿茶醛、丹酚酸B 3个成分的含量。方法:采用AgilentZorbax SB-Cl8(3.0 mm×100 mm,3.5μm)色谱柱,以乙腈-0.5%醋酸为流动相进行梯度洗脱,流速0.5 mL.min-1,检测波长288 nm。结果:丹参素钠、原儿茶醛、丹酚酸B浓度分别在76.84～1921μg.mL-1(r=0.9996)、9.624～240.6μg.mL-1(r=0.9999)、10.08～252.0μg.mL-1(r=0.9994)范围内呈良好的线性关系;上述3个成分精密度试验的RSD<1%,48 h内稳定性试验的RSD<2%,加样回收率试验的RSD<2%。结论:本文方法简便可靠,能同时测定丹香冠心注射液中丹参素钠、原儿茶醛、丹酚酸B 3个有效成分的含量,可用于丹香冠心注射液的质量控制。     

RP-HPLC测定血清中氧氟沙星的浓度

目的建立测定血清中氧氟沙星浓度的方法。方法采用反相液相色谱-荧光法。固定相为Phenomenex C18柱(5μm,4.6 mm×250 mm);流动相为甲醇-0.05 mol.L-1磷酸二氢钠溶液p、H2.3(35∶65);检测波长λex=298 nm,λem=491 nm;流速1.0ml.min-1。结果氧氟沙星的线性范围为0.037~4.064μg.ml-1(r=0.9997);方法回收率及提取回收率分别为97.3%和92.4%;其日内精密度RSD<3.2%,日间精密度RSD<5.4%。结论所用方法准确、简便、重复性好,适用于生物样品中氧氟沙星的浓度测定。     

LC-MS/MS测定人血浆中的辛伐他汀

目的建立测定人血浆中辛伐他汀的方法。方法采用LC-MS/MS法,血浆样品中加入内标洛伐他汀,经乙腈沉淀蛋白后,以正离子多反应监测(MRM)方式测试,用于定量的离子为m/z441.2→325.2(辛伐他汀)和m/z427.2→325.2(洛伐他汀)。色谱柱为Restek Allure C18(50 mm×2.1 mm,5μm),流动相为乙腈-0.5%甲酸水溶液(70:30)。结果血浆中辛伐他汀的线性范围为0.1-50.0 ng.ml-1,定量限为0.1 ng.ml-1,方法回收率为98%~102%,日内RSD<3.6%,日间RSD<5.6%。结论所建方法专属性好、灵敏度高,符合血浆样品测定的要求,适用于辛伐他汀的临床药物动力学研究。     

柱切换HPLC测定血浆中消旋卡多曲的活性代谢物thiophine

目的建立柱切换RP-HPLC测定消旋卡多曲的活性代谢物(th ioph ine,TP)浓度的方法。方法采用Krom asil C18预柱(25 mm×4.6 mm,5μm)和分析柱(100 mm×4.6 mm,5μm),流动相均为0.02 mol.L-1磷酸二氢钾-乙腈(80:20);取已加抗氧剂的血浆0.5 m l,加入pH 2.0的磷酸盐缓冲液酸化,经C18SPE柱萃取后进样,在210 nm处检测,按外标法定量。结果标准曲线在25.0~3.2×103μg.L-1范围内有良好的线性,最低定量限为25μg.L-1,TP的保留时间为4.7 m in,日内RSD<8%,日间RSD<9.5%,方法回收率95%~115%。结论所建方法快速简便,灵敏准确,适用于TP血药浓度的测定及卡多曲的药物动力学、生物利用度的研究。     

柱切换HPLC测定血浆中消旋卡多曲的活性代谢物thiophine

目的建立柱切换RP-HPLC测定消旋卡多曲的活性代谢物(th ioph ine,TP)浓度的方法。方法采用Krom asil C18预柱(25 mm×4.6 mm,5μm)和分析柱(100 mm×4.6 mm,5μm),流动相均为0.02 mol.L-1磷酸二氢钾-乙腈(80:20);取已加抗氧剂的血浆0.5 m l,加入pH 2.0的磷酸盐缓冲液酸化,经C18SPE柱萃取后进样,在210 nm处检测,按外标法定量。结果标准曲线在25.0~3.2×103μg.L-1范围内有良好的线性,最低定量限为25μg.L-1,TP的保留时间为4.7 m in,日内RSD<8%,日间RSD<9.5%,方法回收率95%~115%。结论所建方法快速简便,灵敏准确,适用于TP血药浓度的测定及卡多曲的药物动力学、生物利用度的研究。     

HPLC测定氧氟沙星麻黄碱滴鼻液的含量

目的建立测定氧氟沙星麻黄碱滴鼻液含量的方法。方法采用HPLC法同时测定滴鼻液中氧氟沙星与麻黄碱的含量。色谱柱为AlltimaC18(250mm×4.6mm,5μm);柱温35℃;流动相为0.05mol.L-1醋酸铵溶液(用三乙胺调节pH4.0)-乙腈(85∶15);流速1.0ml.min-1;检测波长254nm;检测灵敏度为0.1AUFS,进样量20μl。结果氧氟沙量在1.5μg.ml-1~0.75mg.ml-1浓度范围内,峰面积与浓度的线性关系良好(r=0.9999),平均回收率为101.2%;麻黄碱在5μg.ml-1~2.5mg.ml-1范围内,峰面积与浓度的线性关系良好(r=0.9999),平均回收率为101.4%;重复性试验的RSD=0.5%(n=9)。结论所用方法快速、简便,精密度好,灵敏度高,可作为氧氟沙星麻黄碱滴鼻液的含量控制方法。     

一阶导数分光光度法测定头孢拉定胶囊含量

目的探讨头孢拉定胶囊中头孢拉定的含量测定方法。方法利用一阶导数分光光度法,在头孢拉定对照品波峰(247 nm)之间辅料一阶导数接近零,对本品测定没有干扰。结果头孢拉定浓度在10~70μg.ml-1范围内线性关系较好,平均回收率为99.7%,相对标准偏差为0.18%。结论本法简便、快速、经济,适用于大生产过程中间体的控制。     

人体内阿昔洛韦血药浓度的HPLC测定

目的:建立阿昔洛韦血药浓度测定方法,用于阿昔洛韦缓释片血药浓度研究。方法:采用高效液相色谱法。分析柱为C1 8柱,流动相为2 %乙腈水溶液(p H3) ,检测波长2 5 4 nm。结果:样品组分在0 .0 5～1.0μg·ml- 1 线性良好(0 .9997) ,最低血浆药物检测浓度为0 .0 1μg·ml- 1。回收率为92 .36 %～99.2 1% ,日内及日间RSD<10 %。结论:本法简便、迅速、灵敏、准确,适合于临床药学研究需要。     

HPLC测定银杏叶中聚戊烯醇的含量

目的建立测定银杏叶中聚戊烯醇含量的方法。方法采用HPLC法,色谱柱为Kromasil ODS柱(250 mm×4.6 mm,5μm),流动相为异丙醇-甲醇-正己烷-水(50∶25∶15∶2),流速0.9 ml.min-1,柱温27℃,检测波长210 nm。结果在1.08~2.88μg.ml-1范围内,聚戊烯醇浓度与峰面积呈良好的线性关系(r=0.9979)。平均回收率为96.8%(RSD<2%,n=3)。结论所建方法快速、准确、稳定,为银杏叶中聚戊烯醇含量测定提供了可靠的方法。     

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.