UPLC-MS/MS法测定人血浆中双氯芬酸钠的浓度

目的建立测定人血浆中双氯芬酸钠浓度的超高效液相色谱-串联质谱(LC-MS/MS)检测方法。方法采用蛋白沉淀法处理血浆样品。色谱柱为Waters C18(50 mm×2.1 mm,1.7μm),流动相为乙腈-体积分数为0.06%的甲酸溶液(体积比为40∶60),流速为0.7 m L·min-1,正离子模式多反应监测(MRM)扫描分析,离子通道分别为m/z 296.03→249.98(双氯芬酸钠),m/z 358.10→138.92(吲哚美辛,内标物)。结果血浆中双氯芬酸钠线性为10³ 000μg·L-1(r=0.998 0),定量下限为10μg·L-1,提取回收率在95.1%3 000μg·L-1(r=0.998 0),定量下限为10μg·L-1,提取回收率在95.1%¹01.3%,日内、日间精密度RSD均小于10%。结论该方法可用于双氯芬酸钠的药物动力学研究。     

超快速液相色谱-质谱/质谱联用法测定人血浆中兰索拉唑的浓度

目的 建立测定人血浆中兰索拉唑(抗胃及十二指肠溃疡药)含量的超快速液相色谱-质谱/质谱联用法(UFLC-MS/MS).方法 血浆样品经甲醇沉淀蛋白后,以乙腈-0.1%甲酸(27:73)洗脱,用Ultimate XB-CN(5μm,2.1mm×150mm)色谱柱,通过电喷雾离子化三重四级杆串联质谱,经多反应监测模式检测,m/z370.4→m/z252.2(兰索拉唑),m/z 237.2→m/z 194.3(卡马西平,内标).结果 兰索拉唑在5.01～5007.90μg·L-1内线性关系良好(γ=0.998 5),最低定量浓度为5.01μg·L-1.方法 回收率(n=5)分别为90.21%,89.71%,92.27%,日内和日间精密度均<9%.结论 该法操作简单,灵敏,准确,重现性好,适用于该药的临床药代动力学研究.     

高效液相串联质谱法测定大鼠血浆中咪哒唑仑的浓度

目的:建立测定大鼠血浆中咪哒唑仑浓度的液相色谱串联质谱法。方法:以伊曲康唑为内标,色谱柱为XTerra MSC18(150mm×2.1mm,3.5μm);流动相为乙腈-0.02%醋酸铵(75∶25);流速为0.2mL.min-1;质谱仪为电喷雾-三重四极杆质谱,以多反应监测(MRM)方式对咪哒唑仑(m/z326.2→m/z291.2)和伊曲康唑(m/z705.3→m/z392.1)进行测定。结果:咪哒唑仑在125～50 000μg.L-1范围内线性关系良好(r=0.996 8),最低检测限为5.0μg.L-1。低、中、高浓度的平均回收率为97.22%,101.88%,90.57%;日内及日间精密度均小于10%。结论:本方法简单、快速、灵敏,能有效检测大鼠血浆中咪哒唑仑的浓度。     

HPLC-MS/MS法测定人血浆中甲氧氯普胺的浓度

目的建立测定人血浆中甲氧氯普胺浓度的LC-MS/MS方法。方法采用Alltech Alltima C18色谱柱(2.1 mm×50 mm,3μm),以5 mmol.L-1乙酸铵-乙腈-甲醇(体积比为50∶40∶10)为流动相,流速为0.2 mL.min-1,血浆样品在碱性条件下液-液萃取,通过电喷雾离子化四极杆串联质谱,以多离子反应监测(MRM)方式进行检测。用于定量分析的离子对分别为m/z300.20→m/z227.00(甲氧氯普胺)和m/z152.20→m/z134.00(内标,苯丙醇胺)。结果甲氧氯普胺线性范围为0.5~200.0μg.L-1,最低定量限为0.5μg.L-1,平均回收率为(71.38±6.35)%,日内和日间精密度均小于15%。结论该法适用于临床药物浓度监测和药物动力学的研究。     

液相色谱-串联质谱法测定人血浆中氟氯西林的含量

目的为临床制定安全有效的给药方案提供参考。方法采用液相色谱-串联质谱(LC-MS-MS)法。血浆样品经液-液萃取处理后,以乙腈-水(体积比25∶75)为流动相,采用Zorbax Ec lipseXDB C8柱(150 mm×4.6 mm,5μm)分离;通过电喷雾三重四级杆串联质谱,以负离子、选择反应监测方式进行检测,用于定量的离子反应分别为m/z452→m/z311(氟氯西林)和m/z468→m/z327(内标双氯西林)。结果氟氯西林线性范围为20.0~15000μg.L-1,定量下限为20.0μg.L-1,提取回收率均大于70%,日内、日间精密度均小于7%。结论该法适用于注射用氟氯西林钠的单次肌内注射给药的药物动力学研究。     

UPLC-MS/MS法测定比格犬血浆中阿司匹林、水杨酸和单硝酸异山梨酯的浓度

目的:建立灵敏的超高效液相色谱-质谱联用法测定比格犬血浆中的阿司匹林、水杨酸和单硝酸异山梨酯的浓度。方法:选用Waters BEH C18(100 mm×2.1 mm,1.7μm)色谱柱,分别以0.5%甲酸/5 mmol.L-1醋酸铵-乙腈和2 mmol.L-1醋酸铵-乙腈为流动相,采用梯度洗脱进行分离,样品采用乙酸乙酯提取后进样,通过电喷雾电离源,以多重反应监测(MRM)方式进行负离子检测,用于定量分析的离子对分别为m/z 178.9→136.9(阿司匹林)、m/z 136.9→64.9(水杨酸)和m/z 293.7→250.0(内标,双氯酚酸钠)测定阿司匹林和水杨酸;m/z 249.6→58.9(单硝酸异山梨酯)和m/z 293.7→250.0(内标,双氯酚酸钠)测定单硝酸异山梨酯。结果:阿司匹林、水杨酸和单硝酸异山梨酯的线性范围分别为2.0～2000.0,20.0～16000.0,9.6～9557.0 ng.mL-1;定量下限分别可达2.0,20.0,9.6 ng.mL-1;日内、日间精密度(RSD)均小于15%。结论:本方法灵敏度较高,血浆用量少,适用于比格犬血浆样品中阿司匹林、水杨酸和单硝酸异山梨酯的测定和药物动力学研究。     

LC-MS/MS法测定大鼠血浆中阿魏酸的血药浓度

目的建立高效液相色谱-串联质谱(LC-MS/MS)法测定大鼠血浆中阿魏酸的血药质量浓度。方法色谱柱采用Diamonsil C18柱(150 mm×4.6 mm,5μm),仪器采用API 4000三重四极杆串联质谱仪,采用多反应监测(multiple reaction monitoring,MRM)扫描方式检测,采用沉淀蛋白法处理样品,用于定量分析的离子对为m/z 193.1→m/z 134.1(阿魏酸)和m/z 121.2→m/z 77.0(苯甲酸,内标)。结果该方法的线性范围在5.0～2 000.0μg.L-1,定量下限为5.0μg.L-1,且血浆中内源性物质不干扰阿魏酸的测定;日内和日间精密度RSD均小于15%;阿魏酸的平均提取回收率为91.0%～100.5%,基质效应在87.8%～89.7%。结论该方法适用于阿魏酸在大鼠体内的药物动力学研究。     

LC-MS/MS方法同时检测人血浆中异烟肼、乙胺丁醇和吡嗪酰胺的浓度

目的:建立同时检测人血浆中3种抗结核药物异烟肼、乙胺丁醇和吡嗪酰胺浓度的LC-MS/MS方法,用于肺结核患者及临床试验中三药血药浓度的测定。方法:以对乙酰氨基酚为内标,血浆样品经乙腈沉淀蛋白等处理后检测。采用AgilentZORBAX SB-Aq色谱柱(2.1 mm×100 mm,3.5μm)为分析柱,ZORBAX SB-Aq柱(2.1 mm×12.5 mm,5μm)为保护柱,以乙腈-5 mmol·L-1甲酸铵水溶液(含0.1%甲酸)(8:92,v/v)为流动相,使用电喷雾离子源(ESI),以正离子多反应监测(MRM)方式进行检测,异烟肼m/z 138.2→121.0,乙胺丁醇m/z 205.2→116.1,吡嗪酰胺m/z 124.1→81.1,对乙酰氨基酚m/z152.0→110.0。分析时间为5 min。结果:血浆中内源性物质对测定无干扰,异烟肼线性范围为0.1～6.0μg.mL-1,定量下限(LLOQ)为0.1μg.mL-1,乙胺丁醇线性范围为0.1～5.0μg.mL-1,定量下限(LLOQ)为0.1μg.mL-1,吡嗪酰胺线性范围为1.0～50.0μg.mL-1,定量下限(LLOQ)为1.0μg.mL-1。日内、日间精密度(RSD)均小于10%,准确率为90.4%～108.7%。结论:本方法特异性强,灵敏度高,测定结果可靠,适用于临床血浆样品的高通量分析。     

高效液相色谱-质谱串联法同时测定人血清中拉莫三嗪及卡马西平的含量

目的:建立高效液相色谱-质谱串联(LC-MS/MS)法测定人血清中拉莫三嗪及卡马西平的含量。方法:取人血清经乙腈直接沉淀后进样分析,色谱柱为Diamonsil C18(150 mm×4.6 mm,5μm),流动相为甲醇∶0.2%甲酸水溶液,流速为0.4 mL.min-1;流动相采用梯度洗脱。质谱采用ESI离子源,正离子多反应监测扫描分析,拉莫三嗪离子对为m/z 256→145,卡马西平离子对为m/z 237→194。结果:卡马西平和拉莫三嗪血药浓度分别在5～500μg.L-1(r=0.998 9)和5～500μg.L-1(r=0.997 2)范围内线性关系良好,定量下限分别为5μg.L-1。日内、日间RSD均<7.0%;提取回收率均>85%。结论:本方法处理样品简便,适用于临床血药浓度监测及为联合用药的药动学研究奠定基础。     

电喷雾离子阱二级质谱法测定人血浆中利培酮浓度

目的 建立电喷雾离子阱二级质谱法(LC-ESI-MS/MS)测定人血浆中利培酮浓度.方法 色谱柱DiamonsilTM C18(4.6mm×150mm,5μm);流动相:乙腈(1%甲酸):0.02mol·L-1醋酸铵=(60:40,V:V),流速:0.8 mL·min-1;柱温:25℃;进样体积:10μL;质谱条件为电喷雾离子源,检测方式为正离子多离子反应监测(MRM),用于定量分析的离子为利培酮m/z411→191,内标替米沙坦m/z516→497,生物样本采用醋酸乙酯:二氯甲烷(4:1)液液萃取处理.结果 利培酮血药浓度线性范围为0.5～50 μg·L-1,线性方程为C=2.38F-0.24,r=0.999(n=7),最低检测浓度为0.1μg·L-1,高中低三个浓度的提取回收率分别为71.70%,64.58%,64.93%,日内、日间RSD均小于15%.结论 本法灵敏、准确、快速,可用于临床常规血药浓度测定和药动学研究.     

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.     

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.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

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.