高效液相色谱法同时测定人血浆中多种喹诺酮类药物浓度

目的建立一种可同时测定人血浆中多种喹诺酮类药物浓度的高效液相色谱法。方法以Kromasil C8柱(250 mm×4.6 mm,5μm)为色谱柱,含1%三乙胺的0.02 mol/L磷酸二氢钾缓冲液(用磷酸调pH至3.0)-甲醇-乙腈(65∶20∶15)为流动相,流速1.0 mL/min,柱温30℃,检测波长依受检药物而定,进样量20μL,不同喹诺酮类药物互为内标。采用二氯甲烷以液液萃取法提取样品,氮气流吹干后以流动相复溶进样。结果在上述色谱条件和样品处理条件下,加替沙星、莫西沙星等几种喹诺酮类药物可得到有效分离和检测。以莫西沙星为模型药物,对建立的测定方法进行验证,莫西沙星在血药质量浓度0.05～5.0μg/mL范围内线性关系良好(r=0.999 8),最低定量限为0.05μg/mL;高、中、低质量浓度方法回收率为99.37%～104.50%,提取回收率为80.48%～91.25%;日内精密度RSD为4.03%～5.73%,日间精密度RSD为4.53%～6.83%。样品在低温冷冻(-20℃)条件下至少1个月内稳定。结论所建立的高效液相色谱法准确可靠、简便快速、灵敏度较高,可用于莫西沙星等多种喹诺酮类药物的血药浓度测定。     

反相高效液相色谱法测定人血浆中利鲁唑浓度

目的 :建立测定利鲁唑血药浓度的方法。方法 :采用反相高效液相色谱法 ,以DiamonsilTM C18(250mm×4. 6mm ,5μm )为色谱柱 ,甲醇 -0 .03mol/L磷酸二氢铵 (80∶20 ,V/V)为流动相 ,乙醚为提取剂 ,流速为0. 8ml/min ,检测波长为265nm。结果 :利鲁唑检测浓度线性范围为5～1000ng/ml ,最低检测浓度为5ng/ml ;高、中、低3种浓度的回收率分别为99 .51 %、95. 74 %和97. 12 % ,日内RSD≤1 .17 %、日间RSD≤6. 48 % (n=5)。结论 :本方法灵敏、准确、简单、快速 ,可用于人体药动学研究。     

饲料对大鼠尿液中草酸含量的影响

目的建立HPLC法测定大鼠尿液草酸的含量,研究大鼠饮食对其尿液中草酸含量的影响。方法采用AgilentSB C18(250 mm×4.6 mm,5μm)色谱柱;Agilent zorbax extend-C18(4.6 mm×12.5 mm,5μm)为保护柱;以0.018 mol.L-1磷酸二氢钾(含0.0025 mol.L-1四丁基硫酸氢铵,磷酸调pH 2.5~2.7)为流动相;检测波长210 nm;流速1.5 ml.min-1;柱温25℃;进样量20μl;以50 mg.ml-1磺基水杨酸沉淀尿蛋白。观察不同的饮食条件对大鼠尿液草酸含量的影响。结果最低检测限为0.4μg.ml-1。线性范围为62.5~4000μg.ml-1,平均回收率为103.49%,日内及日间精密度RSD<11.56%。在不同的饮食条件下,大鼠尿液草酸含量的波动明显。结论该方法简便,灵敏度高,重复性好,可用于尿液草酸钙结石成因的研究。     

反相高效液相色谱法测定清肝助眠汤中芍药苷的含量研究

目的建立以反相高效液相色谱(RP-HPLC)法测定清肝助眠汤中芍药苷含量的方法。方法色谱柱Agilent Technologies C18(250mm×4.6mm,5μm),流动相为甲醇-1%磷酸(30∶70),流速为1.0ml/min,柱温为25℃,检测波长为230nm。结果芍药苷检测量在0.34～0.17μg范围内与峰面积有着线性关系(r=0.9999);平均加样回收率为100.04%,RSD=0.78%(n=9)。结论该方法方便易行,结果准确,重复性好。     

高效液相色谱法测定人血清中氧氟沙星浓度

本文建立了反相高效液相色谱法测定人体血清氧氟沙星浓度的方法。该法用Zorbax-ODS(5μ)色谱柱,0.03mol/L 磷酸;乙腈:40%氢氧化四丁铵水溶液(80∶7∶1,v/v)的混合溶液(pH≈2.3)为流动相,荧光检测器(激发波长295nm,发射波长467nm)检测,吡哌酸为内标物。最低检测浓度为1ng/ml,回收率为91.4%～102.8%。     

RP-HPLC测定洛伐他汀及胶囊的含量

目的 采用HPLC测定洛伐他汀及其胶囊的含量.方法 色谱柱为Agela C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.01%磷酸(60:40),流速1.0 ml·min-1.检测波长238 nm.结果 洛伐他汀的线性方程为:A=5.980×104+2.910×104C(r=0.9999),线性范围4.88～305.00 μg·ml-1,胶囊平均回收率为100.20%,RSD=0.24%(n=9).结论 所建方法准确、重复性好、操作简单,可有效地控制药品质量.     

反相高效液相色谱法测定人血浆中盐酸二甲双胍浓度

目的:建立测定人血浆样品中盐酸二甲双胍浓度的定量分析方法.方法:反相高效液相色谱法(RP-HPLC法).色谱柱为Nucleosil 100-5硅胶柱(250 mm×4.6 mm,5 μm),流动相为0.03 mol/L的磷酸二氢铵溶液(pH=7.00)-乙腈(75:25,V/V),检测波长240 nm,柱温30℃,进样量40μL.结果:血浆中盐酸二甲双胍浓度线性范围是25～4 000 ng/mL(r=0.999 9),回收率在94.59%～100.84%之间,日内RSD(n=5)在0.67%～2.35%之间,日间RSD(n=5)在2.77%～4.09%之间,最低检测限是12.5 ng/mL.结论:HPLC法简便、快捷、灵敏、准确,适用于临床药代动力学及药效学的研究.     

HPLC法测定阿莫西林/舒巴坦的含量

目的:建立同时测定阿莫西林和舒巴坦含量的方法,为进一步考察在不同输液中阿莫西林和舒巴坦稳定性提供检测依据。方法:采用高效液相色谱法,色谱柱为C18柱(250mm×4.6mm,7μm);流动相:5mmol/L四丁基氢氧化铵溶液(1mol/L磷酸溶液调pH=5.0):乙腈(65∶35),流速:1.0ml/min,检测波长为230nm。结果:阿莫西林和舒巴坦分别在5.0～160μg/mL和2.5～80μg/mL检测浓度范围范围内呈良好的线性关系,相关系数为0.9999和0.9997,平均回收率为95.5%和95.4%,RSD%分别为1.26和1.18。结论:该方法能简便、快速、准确地同时测定输液中阿莫西林和舒巴坦的含量。     

反相HPLC法测定阿克拉霉素含量

本文建立了反相高效液相色谱法分离测定阿克拉霉素含量,采用C8,5μm,4.0×125mm柱(Lichrospher).流动相为含水0.1%三乙胺的0.005M乙二胺四醋酸二钠溶液(醋酸调节pH为4.5)-乙腈(66:34),流速为2ml/min.检测波长为432nm.线性范围0.05～1mg/ml,r.=0.9996(n=5).阿克拉霉素平均回收率为100.7%(n=3).RSD为0.8%(n=6).     

反相高效液相色谱法测定GCLE含量及有关物质

目的 建立反相高效液相色谱法测定GCLE的含量及检查有关物质.方法 ODS-C18色谱柱(150mm×4.6mm,5μm);以乙腈:四氢呋喃:水(用1mol/L磷酸调pH3.5)(45:15:40,v/v/v)为流动相;检测波长为254nm;流速1.0ml/min;柱温25℃.结果 GCLE进样浓度在0.2537～1.0150mg/ml范围内,浓度与峰面积呈良好的线性关系(r=0.9998),日间精密度RSD=0.17%(n=6).结论 该方法简便、快速、准确且重现性好,可作为GCLE含量测定及有关物质控制方法.     

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.