HPLC／MS测定人血浆中盐酸班布特罗及其代谢物特布他林的浓度

目的 :用高效液相 /质谱联用法测定人血浆中盐酸班布特罗及其代谢物特布他林的浓度。方法 :液相 :采用SupelcodiscoveryC18色谱柱 (5 μm ,2 5 0mm× 4 6mm) ;柱温 40℃ ;流动相为甲醇 -醋酸铵溶液 (0 0 0 7mol·L-1) (2 0∶80 ) (并用冰醋酸调 pH =4 8) ,流速 0 6mL·min-1,进样量 6 0 μL ;质谱 :大气压化学电离源 (APCI) ,选择性监测 (SIR)质荷比 (m/z)分别为 2 2 6 (特布他林 ) ,2 6 0 (内标 ) ,36 8(班布特罗 )带正电荷的分子离子峰定量。样品用固相萃取小柱提取处理。结果 :班布特罗线性范围 0 12 5～ 16 μg·L-1,最低检测浓度为 0 0 5 μg·L-1。特布他林线性范围 0 312 5～ 40 μg·L-1,最低检测浓度为 0 0 5 μg·L-1。班布特罗和特布他林的萃取回收率均在 90 %以上 ,日内、日间的RSD皆小于 15 %。结论 :适用于临床上测定血浆中盐酸班布特罗及其代谢物特布他林的浓度及药动学的研究     

高效液相色谱法同时测定血浆中艾司唑仑、三唑仑和阿普唑仑浓度

目的 :建立同时测定血浆中艾司唑仑、三唑仑和阿普唑仑浓度的方法。方法 :采用HPLC法 ,选用ZORBAXRP C18柱(15 0mm× 4 .6mm ,5 μm) ;甲醇 2 5mmol·L-1醋酸铵溶液 (5 7∶4 3)作流动相 ;检测波长为 2 30nm。结果 :本法对三唑仑、阿普唑仑和艾司唑仑的最低检测限浓度均为 4 μg·L-1;回收率接近 10 0 % ;日内、日间精密度 <5 % ;线性范围均为 2 0～ 10 0 0 μg·L-1。结论 :本法能同时测定血浆中三唑仑、阿普唑仑和艾司唑仑 ,此法重现性好 ,灵敏 ,可靠 ,可用于临床药物中毒监测。     

GC-ECD法检测血浆中地西泮及去甲西泮

目的用GC-ECD法分析血浆中地西泮及其主要代谢物去甲西泮。方法血浆中加入内标去烃基氟西泮,调pH至10.8,用二氯甲烷-正己烷(7∶3)提取地西泮及去甲西泮,用GC-ECD法进行分析。结果检材中分析物的回收率80%以上,检出限5μg·L-1以下。结论该方法灵敏度高,可用于口服地西泮10mg人体24h内血浆的分析。     

高效毛细管电泳法测定班布特罗及其代谢物特布他林的血药浓度

目的 :建立测定人血浆中班布特罗及其代谢物特布他林血药浓度的高效毛细管电泳方法。方法 :运行缓冲液为 80mmol·L-1磷酸二氢钠缓冲液 (用磷酸调 pH =3 15 ) ;电压进样 :10kV ,10s ;运行电压 2 5kV ,检测波长为 2 0 0nm ,采用固相萃取的方法进行班布特罗及其代谢物特布他林的提取。结果 :班布特罗及其特布他林峰形良好 ,且血浆组分无干扰峰 ,采用普萘洛尔为内标 ,班布特罗测定的线性范围为 16～ 0 12 5 μg·L-1,特布他林测定的线性范围为 40～ 0 312 5 μg·L-1。本法的日内RSD小于 11% ,日间RSD小于 12 %。结论 :本法是一种简单、灵敏的高效毛细管电泳柱上富集方法 ,适用于药代动力学及其生物等效性的研究     

高效液相色谱法测定 地西泮中毒患者血浆与尿中地西泮浓度

［摘要］目的建立高效液相色谱法测定血浆与尿中地西泮浓度。方法采用Agilent Eclipse XDB C18色谱柱(4.6 mm×150 mm, 5 μm )，以甲醇 水（65：35）为流动相，流速1.0 mL·min-1，检测波长254 nm。结果地西泮和内标物出峰时间分别为3.5 和7.3 min，分离效果较好；血浆样品中地西泮3种浓度（4.0，8.0，16.0 μg·mL 1）日内和日间RSD均＜6%，尿样中地西泮3种浓度（4.0，8.0，16.0 μg·mL 1）日内和日间RSD均＜5%；血浆中地西泮回收率为95.63%～98.33%，尿样中地西泮回收率为88.33%～95.33%，血清及尿样样品线性范围为2.0～64.0 μg·mL 1，相关系数0.995 8。血浆、尿样品地西泮最低检出限分别为2.0及1.5 μg·mL 1。结论该条件可对地西泮中毒患者进行药物快速检测并定量分析。     

高效液相色谱法测定克林沙星血药浓度

目的 :建立用高效液相色谱法测定血浆中克林沙星浓度的方法。方法 :血浆样品在酸性条件下 ,以Oasis小柱提取环丙沙星为内标 ,采用LichrosorbC18(5 μm)柱 ,流动相为乙腈 - 0 0 5mol·L-1柠檬酸三乙胺溶液 (pH 2 5 ) (2 0∶80 ) ,流速为 1 0mL·min-1,检测波长 30 0nm ,克林沙星和内标的保留时间分别为 7 1min和 4 5min。结果 :线性范围在 0 0 3～10 μg·mL-1(r=0 9998) ,最低检测浓度为 0 0 2 μg·mL-1,RSD <7%。用本法测定了 6只大鼠灌胃剂量 5 0mg·kg-1克林沙星后血浆中克林沙星的浓度经时变化过程。结论 :本方法可用于克林沙星的血药浓度测定及药代动力学研究。     

反相离子对高效液相色谱法测定血浆吗啡及其代谢物M3G的含量

目的:建立反相离子对高效液相色谱法测定血浆吗啡及其代谢物3-葡萄糖醛酸吗啡(M3G)的浓度。方法:血浆样品以乙酸乙酯提取,以纳洛酮为内标。应用HPLC-UV方法测定吗啡浓度,分析柱DiamonsilTMC18(250mm×4.6mm),流动相为0.01mol·L-1磷酸二氢钾缓冲液(含0.5mmol·L-1的十二烷基磺酸钠)-乙腈(71:29),检测波长210nm,流速0.65ml·min1。血浆样品经β-葡萄糖醛酸苷酶水解测定其代谢物含量。结果:吗啡标准曲线线性范围宽(10-4000μg·L-1),线性关系良好;最低检测限10μg·L-1;高、中、低浓度的回收率在85·3%以上。血浆M3G在250-8250μg·L-1浓度范围内与水解增加吗啡浓度呈线性关系。结论:本法简单,快速,血浆中杂质不干扰样品的测定,满足生物样品分析要求。     

气相色谱-电子捕获法测定人血中舍曲林及其代谢产物去甲舍曲林的浓度

目的 :建立一种测定人血浆中舍曲林及其代谢产物去甲舍曲林浓度的方法。方法 :在碱性条件下用乙醚 正己烷 (80∶2 0 ,V/V)一次萃取 ,并以 10 %的三氟醋酐在 5 0℃下进行衍生化处理。采用 5 %苯基石英毛细管柱 (30m× 0 .2 5mm ,0 .2 5 μm) ,柱温 2 37℃。高纯度N2 为载气 ,流速 0 .75mL·min-1,分流比 2 5∶1,柱前压 115kPa。进样口温度 2 6 0℃ ,63 Ni电子捕获检测器 ,检测器温度 30 0℃。结果 :舍曲林和去甲舍曲林的线性范围均为 1.0～ 10 0 μg·L-1,最低检测浓度 0 .5 μg·L-1,相对回收率分别为96 .8%～ 10 8.2 %和 94 .5 %～ 10 6 .7% ,日内和日间RSD <8.5 %。结论 :本法简便、准确、灵敏 ,重现性好 ,是一种有效的检测人血浆中舍曲林及去甲舍曲林浓度的方法     

HPLC-MS法测定人血浆中米氮平的浓度

目的 :建立一种用高效液相色谱 -电喷雾离子化质谱 (HPLC -MS)联用技术测定米氮平血药浓度的方法。方法 :以0 0 1mol·L-1乙酸铵水溶液∶甲醇 (2 7∶73,V/V)为流动相 ,盐酸地尔硫 艹卓 为内标 ,血浆样品经乙酸乙酯萃取后上样 ,经C18柱分离后 ,以质谱为检测器 ,采用选择性离子检测 (SIM)测定人体血浆中米氮平的浓度。结果 :米氮平的线性范围 0 2 0～ 15 0 μg·L-1(r=0 9995 ) ,平均相对回收率在 90 %～ 110 %之间 ,日内和日间RSD均 <5 % ,米氮平的最低定量限为 0 2 0 μg·L-1,提取回收率 >85 %。结论 :该方法快速、准确、灵敏 ,可用于米氮平的药动学研究     

固相萃取-高效液相色谱法同时测定血浆中氯米帕明和4种苯二氮类药物浓度方法的研究

目的建立同时测定血浆中氯米帕明和4种苯二氮类药物奥沙西泮、地西泮、氯硝西泮、阿普唑仑浓度的固相萃取-高效液相色谱法(HPLC)。方法 XTerra C8 RP(4.6mm×150 mm,5μm)为色谱柱,磷酸盐缓冲液(50 mmol·L-1,p H 3.0)和乙腈(73.2:26.8,V/V)为流动相,流速1.2m L·min-1,柱温45℃;检测波长220 nm,血浆经C1固相萃取柱预处理。结果线性范围分别为:阿普唑仑、氯硝西泮5.0~200.0μg·L-1,地西泮10.0~500.0μg·L-1,氯米帕明20.0~500.0μg·L-1,奥沙西泮7.5~2 000μg·L-1,相关系数均大于0.999 4;最低检测限分别为:阿普唑仑1.5μg·L-1、氯硝西泮1.4μg·L-1、地西泮3.0μg·L-1、氯米帕明5.5μg·L-1、奥沙西泮2.2μg·L-1;日内及日间精密度(CV%)分别为2.2%~12.6%、2.1%~13.2%,偏差-10.6%~14.6%,提取回收率81.1%~100.1%。结论该法可用于临床血浆中氯米帕明和4种苯二氮类药物的同时检测,方法新颖、灵敏、经济,结果准确、可靠。     

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.     

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.     

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.     

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.