HPLC法测定美罗培南血药浓度及在重症感染患者中的临床应用

目的 建立高效液相色谱(HPLC)法测定人血浆中美罗培南血药浓度,为制定个体化给药方案提供依据。方法 血浆样品经乙腈沉淀蛋白,经过离心后取上清液进样分析;流动相为乙腈∶0.1%三乙胺(65∶35),色谱柱为Agilent Zorbax SB-Aq HILIC(4.6 mm×250 mm, 5μm);柱温30℃;流速1 ml/min;进样量50μl;检测波长299 nm。结果 美罗培南血药浓度在0.35～43.92μg/ml,范围线性关系良好(R²=0.999),定量下限0.35μg/ml,高、中、低浓度的方法回收率(98.76±5.71)%～(109.70±1.96)%;平均提取回收率(96.99±2.97)%～(106.11±2.55)%;批内、批间精密度(RSD)1.98%～7.75%。应用此方法测定13例患者血浆药物浓度并计算患者的f%T_>MIC值,其中有3例患者f%T_>MIC未达标。结论 该方法专属性好、准确、快速、操作简便,适用于美罗培南血药浓度监测,为临床给药方案评价及优化提供重要依据。     

反相高效液相色谱法测定美罗培南的血药浓度及其临床应用

目的建立快速测定人血浆中美罗培南的HPLC法,并用该法对临床使用美罗培南的患者进行血药浓度测定。方法采用蛋白沉淀法对血样进行预处理,以雷尼替丁为内标。色谱条件:Spursil ODS C18柱(250mm×4.6 mm,5μm);柱温:30℃;流动相:甲醇-乙腈混合有机相(10:3)-0.05 mol·L-1磷酸二氢钠缓冲液(15:85,缓冲盐pH=5.0);流速:0.8 mL·min-1;测定波长:298 nm。结果美罗培南的最低定量浓度为0.35μg·mL-1,线性范围为0.35¹81.17μg·mL-1,绝对回收率和相对回收率分别为95.4%和100.0%,日内、日间精密度均<10%。63名患者的血药峰浓度、谷浓度、t>MIC均具有较大差别。结论该方法快捷、经济、稳定、准确,适用于美罗培南临床血药浓度监测。临床患者美罗培南的谷峰浓度差异较大,对患者进行美罗培南峰谷浓度的测定对促进美罗培南在临床的合理应用具有重要意义。     

HPLC法测定人血清中美罗培南浓度

目的:建立HPLC法测定人血清中美罗培南浓度.方法:采用Ko肿asil C18色谱柱(4.6 mm×250 mm,5μm);以甲醇:乙腈:5 mmol·L-1磷酸二氢钾缓冲液(12:10:78,用H3PO4调节pH值至2.8,v/v)为流动相;流速:1.2 ml·min-1;检测波长:298 nm.内标:头孢吡肟.结果:血清中美罗培南的标准曲线方程为:A=0.087 7C 0.005 6,r=0.999 7,在0.5～50 μg·ml-1浓度范围内线性关系良好,平均回收率为100.6%;日内、日间RSD均小于5%.结论:该方法灵敏准确,适用于美罗培南临床药物动力学的研究.     

高效液相色谱法测定人血清和置换液中美罗培南浓度

目的:建立高效液相色谱法(HPLC)测定人血清和置换液中美罗培南浓度的方法。方法:采用HPLC法,以依利特Hypersil ODS2柱(4.6mm×150mm,5μm)为色谱柱;甲醇-0.01mol·L-1磷酸二氢钾缓冲液(9∶91)为流动相,pH4.91;流速为1.4mL·min-1;检测波长为297nm。结果:血清中美罗培南的标准曲线方程为Y=17.027X-4.7541,r=0.9997,在0.5~80mg·L-1范围内线性关系良好,平均回收率为100.2%,日内,日间RSD均小于7.2%;置换液中美罗培南的标准曲线方程为Y=18.931X 0.8928,r=0.9998,在0.5~80mg·L-1范围内线性关系良好,平均回收率99.2%,日内、日间RSD均小于7.6%。结论:该方法灵敏准确,适用于临床药动学的研究。     

美罗培南血清浓度测定及在重症感染患者中的临床应用

目的建立HPLC法测定人血清中美罗培南浓度,应用于重症感染患者治疗药物监测,为其制定个体化给药方案提供参考。方法色谱柱为Wondasil C_(18)柱(4.6 mm×150 mm,5μm),内标为替硝唑,流动相为乙腈:0.015 mol·L~(-1) MOPS溶液(p H=7.3)(梯度洗脱),流速为1.0 mL·min~(-1),检测波长为298 nm,柱温为30℃,进样量为20μL。结果美罗培南血药浓度在0.55~79.09μg·mL~(-1)与峰面积线性关系良好(r=0.9997,n=8);高、中、低3种浓度的方法回收率分别为103.2%、104.6%、98.7%;提取回收率分别为88.28%、90.9%、92.6%;日内与日间精密度RSD均小于5%。应用此方法测定6例美罗培南治疗患者血清谷浓度,有1例患者出现谷浓度小于最低抑菌浓度的情况。结论该方法简便、准确、快速、重现性好,适用于美罗培南血药浓度监测和药动学研究。     

美罗培南的含量测定及与输液配伍的稳定性考察

目的 :建立高效液相色谱法测定美罗培南的含量 ;考察注射用美罗培南与5 %GS、GNS、NS注射液配伍的稳定性。方法 :色谱柱 :Nova -PakC18 柱 ;流动相 :乙腈 -0 05molKH2PO4(4∶96) ;流速 :1 0ml/min ;检测波长 :300nm。取美罗培南加入5 %GS、GNS、NS注射液中 ,在常温条件下观察配伍液的外观变化 ,用HPLC法测定配伍后不同时间美罗培南的浓度 ,同时测定配伍液的pH及不溶性微粒变化。结果 :在12 5～250μg/ml范围内 ,峰面积与浓度呈现良好的线性关系 (r=0 9999) ,平均回收率为99 7 % ,RSD为1 1 %。结论 :本法高效、快速、准确 ;美罗培南在5 %GS、GNS、NS注射液中配伍4h内稳定     

重症患者应用美罗培南的血药浓度监测方法优化及临床应用实例

目的:建立超高效液相色谱-串联质谱(UPLC-MS/MS)法监测重症患者美罗培南血药浓度。方法:以法罗培南为内标,用蛋白沉淀前处理方法。色谱柱为AQ C18(100mm×2.1mm,3μm),流动相为乙腈-0.2%甲酸水溶液(梯度洗脱),流速为0.4mL·min^-1,正离子模式多反应监测(MRM)扫描分析,并考察专属性、标准曲线与定量下限、精密度与回收率、基质效应和稳定性。测定6例使用美罗培南的重症患者血药峰、谷浓度;以%T>MIC作为PK/PD靶目标评估计给药方案。结果:血浆中美罗培南线性范围为0.1~40μg·mL^-1(r=0.999),定量下限为0.1μg·mL^-1,低、中、高浓度的美罗培南提取回收率相一致,日内、日间精密度相对标准偏差(RSD)均小于15%。结论:本方法操作简便快速,特异性强,灵敏度高,可用于临床美罗培南血药浓度的测定及临床美罗培南个体化用药指导。     

高效液相色谱法测定人血清美罗培南浓度

目的 建立人血清中美罗培南的高效液相色谱分析方法.方法 采用Agilent Zorbax C18色谱柱(150 mm×4.6 mm,5μm),流动相为0.1%三乙胺水溶液(用磷酸调节pH为2.8)-乙腈(91 :9),检测波长为298 nm.血清样品用固相萃取.结果 美罗培南血药浓度线性范围为0.50~64.00μg/mL(r=0.9994,n=5),最低检测浓度为0.50μg/mL(S/N>10),方法回收率为100.1%～109.0%(n=5),萃取回收率为68.6%～71.3%(n=5);日内相对标准(偏)差(RSD)为0.2%～4.3%,日间RSD为1.6%～2.4%.结论 该方法简便,准确,灵敏,特异性强,重现性好,可用于血清中美罗培南的测定及人体内药代动力学研究.     

HPLC法同时测定加替沙星和奥硝唑的含量

目的建立同时测定加替沙星及奥硝唑含量的高效液相色谱方法。方法分析柱为Hy-persil-ODSC18柱(250mm×4.6mm,5μm);流动相为0.02mol/L磷酸二氢钾缓冲液(含0.5%三乙胺,用磷酸调pH3.0):甲醇:乙腈(65:18:17);流速:1ml/min;加替沙星、奥硝唑检测波长分别为292nm、318nm;柱温为30℃,进样量20μl。结果加替沙星线性浓度范围为8.0～48.0μg/ml(r=0.9998),日内RSD≤0.52%,日间RSD≤0.92%,平均回收率100.53%;奥硝唑线性浓度范围为8.0～48.0μg/ml(r=0.9999),日内RSD≤0.35%,日间RSD≤0.71%,平均回收率100.56%。结论该方法简便、准确、灵敏,可用于同时测定加替沙星和奥硝唑两组分的含量。     

HPLC测定体液内帕尼培南的浓度

目的建立高效液相色谱法测定人血浆样品及尿样品中帕尼培南浓度的定量分析方法,探讨帕尼培南在人体内药物动力学过程.方法以1mol/L 3-(N-吗啉)丙烷磺酸(pH7.0)为稳定剂,对乙酰氨基酚为内标,在Diamosil(TM)C18柱(5μm,250mm×4.6mm)上,血样测定以甲醇0.04mol/L乙酸铵缓冲液(pH4.0,1090)为流动相,流速1.6ml/min;尿样测定以甲醇0.04mol/L乙酸铵缓冲液(pH4.0,298,)为流动相,流速1.6ml/min,检测波长299.3nm,对帕尼培南进行定量测定.结果血浆中帕尼培南在0.5～50.0μg/ml浓度范围内具良好线性关系(r=0.9999).血浆中三种浓度2.0、10.0、50.0μg/ml平均回收率为98.69%(n=6);三种浓度日内及日间RSD分别为1.92%、2.44%、2.31%(n=5)及2.15%、1.45%、2.68%(n=3).尿中帕尼培南在2～200μg/ml浓度范围内具良好线性关系(r=0.9999),尿中三种浓度10.0、25.0、100.0μg/ml平均回收率为101.30%(n=6);日内、日间RSD分别为2.63%、1.05%、0.88%(n=5)及2.30%、3.85%、2.38%(n=3).结论本方法简便、快捷、灵敏、准确,适用于临床药动学及药效学的研究.     

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.     

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.     

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.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.