盐酸小檗碱单次和多次给药在Beagle犬体内的药动学

目的研究盐酸小檗碱胶囊单次和多次给药后在Beagle犬体内的药动学。方法 6条Beagle犬按150 mg单次和多次口服盐酸小檗碱胶囊,多次给药每天1次,共7 d。采用UPLC-MS/MS色谱法测定犬血浆中盐酸小檗碱的浓度。用DAS 2.0药动学软件处理血药浓度数据。用SPSS统计软件对所得的药动学数据进行显著性差异分析。结果单次给药后主要药动学参数t1/2为(18.85±10.54)h,ρmax为(4.18±2.59)μg.L-1,AUC0-t为(110.04±70.22)μg.h.L-1,AUC0-∞为(121.51±74.19)μg.h.L-1,CL为(1 593.57±745.01)L.h-1,V为(44 509.1±34 995.4)L,tmax为(20.42±22.98)h;多次给药达稳态后主要药动学参数t1/2为(18.53±9.99)h,ρmax为(9.92±7.01)μg.L-1,AUC0-t为(164.51±119.70)μg.h.L-1,AUC0-∞为(172.34±125.03)μg.h.L-1,CL为(1 280.19±709.95)L.h-1,V为(33 655.7±27 632.2)L,tmax为(6.08±4.90)h。结论盐酸小檗碱单次和多次给药后在Beagle犬体内血药浓度均较低,盐酸小檗碱多次给药在Beagle犬体内无明显蓄积现象。     

LC-MS/MS法测定可待因在Beagle犬血浆中的质量浓度

目的建立Beagle犬血浆中可待因质量浓度的液相色谱-串联质谱(LC-MS/MS)测定方法。方法色谱柱为Agela·C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-水-甲酸(体积比为15∶85∶0.5),血浆样品经甲醇沉淀蛋白处理,以多反应监测(multiple reaction monitoring,MRM)扫描方式检测,测定Beagle犬经口给予洛芬待因缓释片后血浆中可待因的质量浓度。结果血浆中可待因质量浓度在0.2²0μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%20μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%¹09.2%,基质效应为119.2%109.2%,基质效应为119.2%¹28.3%。可待因在Beagle犬血浆中主要药动学参数t1/2为(2.9±0.5)h,ρmax为(8.7±3.4)μg·L-1,AUC0-∞为(28.5±7.3)μg·h·L-1。结论该方法适用于可待因在Beagle犬体内药动学的研究。     

LC-MS/MS法测定Beagle犬血浆中阿戈美拉汀的质量浓度

目的建立LC-MS/MS的方法测定Beagle犬血浆中阿戈美拉汀的质量浓度,并应用此方法研究阿戈美拉汀片在Beagle犬体内的药代动力学行为及其相对生物利用度。方法采用:Agela C18色谱柱(150 mm×4.6 mm,5μm I.D),以乙腈-浓度5 mmol·L-1醋酸铵水溶液-甲酸(体积比为58.0∶42.0∶0.1)为流动相,采用沉淀蛋白法,以多反应离子监测(multiple reaction monitoring,MRM)扫描方法进行检测,测定经口给予阿戈美拉汀片后Beagle犬血浆中阿戈美拉汀的质量浓度。结果 Beagle犬血浆中阿戈美拉汀在质量浓度0.50~100μg·L-1内与峰面积呈良好线性关系,相关系数r为0.999 0;日内和日间精密度RSD≤8.6%;阿戈美拉汀的平均提取回收率为90.2%~93.3%,基质效应为104.9%~106.8%;阿戈美拉汀在Beagle犬血浆中主要药动学参数t1/2为(4.2±1.5)h,ρmax为(149±85)μg·L-1,AUC0-24为(293±78)μg·h·L-1,AUC0-∞为(300±78)μg·h·L-1。结论该方法适用于阿戈美拉汀在Beagle犬体内的药代动力学及相对生物利用度的研究。     

RP-HPLC法测定Beagle犬血浆中的烟酸浓度及药动学研究

目的建立反相高效液相色谱法测定犬血浆中烟酸的浓度,并应用到烟酸缓释片在Beagle犬体内的药动学研究。方法采用高氯酸沉淀蛋白处理血浆样本;色谱条件:Megres C18柱(4.6mm×250mm,5μm),流动相:甲醇-10mmol.L-1磷酸二氢钾缓冲溶液(3∶97,V/V),柱温:12℃,流速:0.8mL.min-1,检测波长:263nm;比格犬灌胃给药500mg烟酸缓释片。结果线性范围为0.200~35.0μg.mL-1(r≥0.9996),检测限为50ng.mL-1,方法回收率为98.4%;烟酸缓释片表现出明显的缓释效果。结论方法操作简单、专属灵敏、重现性好,适用于Beagle犬体内烟酸的药动学研究。     

大鼠血浆中ZYY-1的测定方法及其在药动学研究中的应用

目的建立HPLC-UV法测定大鼠血浆中ZYY-1的质量浓度,并将此方法应用于ZYY-1在大鼠体内的药动学研究。方法采用Diamonsil ODS C18色谱柱(200 mm×4.6 mm,5μm),流动相为乙腈-20 mmol·L-1磷酸二氢钾溶液(体积比64∶36,用三乙胺调节pH值为8.0),流速为1.0 mL·min-1,检测波长为242 nm,柱温为30℃。以液-液萃取法处理血浆,测定6只大鼠静脉给予40 mg·kg-1ZYY-1后不同时刻血浆中ZYY-1的质量浓度,采用DAS 2.0药动学软件计算药动学参数。结果 ZYY-1质量浓度在50μg·L-1¹0 mg·L-1内线性关系良好,日内、日间精密度均不大于12.1%。ZYY-1的药动学参数:t1/2α为(2.5±1.8)×10-2h,t1/2β为(0.63±0.53)h,t1/2γ为(36.4±36.0)h,CL为(31.1±10.1)L·h-1·kg-1,V为(9.39±11.2)L·kg-1,AUC0-t为(0.963±0.280)mg·h·L-1。结论该方法可作为大鼠血浆中ZYY-1质量浓度的测定方法,为探讨ZYY-1的药动学行为提供了方法依据。     

尼群地平纳米片在犬体内的药动学与相对生物利用度

目的:建立反相高效液相色谱法(RP-HPLC)测定Beagle犬血浆中尼群地平的浓度,研究尼群地平纳米片在Beagle犬体内的药动学与相对生物利用度.方法:采用RP-HPLC测定12只Beagle犬口服受试药品及参比药品后的经时血药浓度,计算主要药动学参数及相对生物利用度.结果:受试药品与参比药品的主要药动学参数分别是:达峰时间(Tgeak)为(1.38±0.20)h和(1.87±0.22)h,峰浓度(Cmax)为(110.1±14.0)μg·L-1和(98.8±34.2)μg·L-1;曲线下面积(AUC)为(684.1±96.6)μg·L-1·h和(620.7±193.2)μg·L-1·h.结论:尼群地平纳米片吸收较为迅速,可快速发挥降压作用,其生物利用度也有所提高.     

LC-MS/MS法测定Beagle犬血浆中胍法辛的质量浓度

目的建立液质联用(LC-MS/MS)方法测定Beagle犬血浆中胍法辛的药物质量浓度,从而研究胍法辛缓释片在Beagle犬体内的药动学特征。方法色谱柱为Agela C8柱(50 mm×2.1 mm,3μm),流动相为乙腈-水-甲酸(体积比20.0∶80.0∶0.01),采用液-液萃取法,以多反应离子监测(MRM)扫描方式进行检测,测定犬经口给予盐酸胍法辛缓释片后血浆中的药物质量浓度。结果Beagle犬血浆中胍法辛的药物质量浓度在0.05²0.0μg·L-1内线性关系良好,相关系数r=0.9992;日内和日间精密度的RSD≤9.0%;胍法辛的平均提取回收率在94.7%20.0μg·L-1内线性关系良好,相关系数r=0.9992;日内和日间精密度的RSD≤9.0%;胍法辛的平均提取回收率在94.7%¹00.9%内;胍法辛的基质效应在99.9%100.9%内;胍法辛的基质效应在99.9%¹07.5%内;胍法辛在犬血浆中主要药动学参数如下:t1/2为(3.5±1.3)h,ρmax为(3.72±1.37)μg·L-1,tMR 0-24为(4.9±1.6)h,tMR 0-∞为(12.2±11.4)h,AUC0-24为(18.80±4.81)μg·h·L-1,AUC0-∞为(21.63±4.76)μg·h·L-1。结论该方法快速、简便、灵敏,适用于胍法辛在Beagle犬体内的药物动力学研究。     

HPLC-荧光法测定莫西沙星在犬体内的血药浓度及其药动学研究

目的:建立测定犬血浆中莫西沙星浓度的方法,并研究其药动学特性。方法:取Beagle犬6只,单剂量灌服莫西沙星120mg,采用高效液相色谱-荧光法测定给药后72 h内血浆中莫西沙星的浓度,采用DAS软件计算药动学参数。色谱柱为HypersilBDS C18,流动相为乙腈-1.0 mol.L-1醋酸溶液(25∶75,pH=2.0),流速为1.0 mL.min-1,激发波长为273 nm,发射波长为488 nm。结果:莫西沙星检测浓度的线性范围为0.1～20.0μg.mL-(1r=0.999 2),平均相对回收率为89.5%～96.7%,日内和日间RSD均<6%。莫西沙星在犬体内的药动学特性符合二室模型,主要药动学参数cmax:(10.25±4.21)μg.mL-1,tmax:(3.0±1.0)h,t1/2β:(9.76±2.48)h,AUC0～24 h:(142.10±49.80)mg.h.L-1。结论:本方法准确、灵敏、快速,适用于莫西沙星的血药浓度测定和药动学研究。     

液-质联用法同时测定大鼠血浆中的乌头碱、新乌头碱、次乌头碱及其药动学

目的:建立HPLC-MS分析方法同时测定大鼠血浆中的乌头碱、新乌头碱、次乌头碱含量,并用于研究大鼠口服附子煎液后乌头碱、新乌头碱、次乌头碱的药动学。方法:血浆经氨水碱化后用醋酸乙酯进行液-液萃取。色谱柱为Alltima C18柱(250mm×4.6mm,5μm),流动相为甲醇-10mmol.L-1醋酸铵水溶液(75∶25)。质谱检测方式为选择性离子监测,选择监测的离子为m/z646.45(乌头碱),m/z632.38(新乌头碱),m/z615.64(次乌头碱)和m/z336.60(盐酸小檗碱,内标)。结果:血浆中乌头碱在0.05~5μg.L-1、新乌头碱在0.5~50μg.L-1、次乌头碱在2.5~250μg.L-1范围内线性关系良好,定量限为0.05μg.L-1,血浆中的平均提取回收率高于90%,批内和批间精密度均小于15%。结论:本法灵敏、可靠、简便,适用于乌头碱、新乌头碱、次乌头碱的药动学研究。     

盐酸坦洛新缓释胶囊在犬体内的药动学

目的建立犬血浆中坦洛新浓度的测定方法,并应用于盐酸坦洛新缓释胶囊中坦洛新犬体内的药动学研究。方法液相色谱-串联质谱法(LC-MS/MS)测定犬血浆中的坦洛新浓度。坦洛新血浆样品经乙酸乙酯萃取,Agilent ZORBAX SB-C18柱(150 mm×2.1 mm,5μm)分离,阿立哌唑为内标,流动相A为体积分数为0.1%的甲酸水溶液,B为乙腈,线性梯度洗脱,电喷雾电离源(ESI),以多反应离子监测(multiple reaction monitoring,MRM)方式进行正离子检测,用于分析的定量离子分别为m/z409→m/z228(坦洛新),m/z 447.5→m/z 284.8(内标:阿立哌唑)。结果犬血浆中坦洛新的线性为0.1～20.0μg.L-1,定量下限为0.1μg.L-1,日内和日间精密度(RSD)均小于13.59%,准确度(relative error,RE)为-2.54%～4.27%。结论本方法适用于盐酸坦洛新缓释胶囊在犬体内的药动学研究。     

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.