液相色谱-串联质谱法测定人血浆中替扎尼定

目的:建立灵敏的液相色谱-串联质谱法测定人血浆中替扎尼定的浓度。方法:血浆样品碱化后经乙醚液-液萃取,以甲醇-10 mmo·lL-1醋酸铵-甲酸(55∶45∶0.1,v/v/v)为流动相,采用Zorbax SB C18柱(150 mm×4.6 mm,5μm)分离,通过电喷雾电离源,以选择反应监测(SRM)方式进行正离子检测,用于定量分析的离子反应分别为m/z254→44(替扎尼定)和m/z243→210(内标,石杉碱甲)。结果:测定血浆中替扎尼定方法的线性范围为10.0～5000 pg·mL-1,定量下限可达10.0 pg·mL-1。日内、日间精密度(RSD)均小于10.0%,准确度(RE)在±3.6%以内。结论:本方法灵敏度高,血浆用量少,适用于替扎尼定制剂的人体药物动力学研究。     

LC-MS/MS法同时测定人血浆伊曲康唑和羟基伊曲康唑浓度

目的:建立同时测定人血浆伊曲康唑和羟基伊曲康唑浓度的液相色谱-串联质谱法(LC-MS/MS)。方法:100μL血浆样品经液-液萃取后,以乙腈-水-冰醋酸(60:40:1.5)为流动相,经Neucleosil ODS柱(50 mm×2.0 mm,5μm)分离,采用电喷雾电离源,以多反应监测(MRM)方式进行正离子检测。用于定量分析的离子分别为m/z 705→m/z 392(伊曲康唑),m/z 721→m/z 408(羟基伊曲康唑)和m/z 383→m/z 337(内标氯雷他定)。结果:测定血浆伊曲康唑和羟基伊曲康唑的线性范围分别为:10～2 500 ng.mL-1和20～4 000ng.mL-1,最低定量限(LLOQ)分别为:10和20 ng.mL-1。日内、日间精密度(RSD)均<15.0%,准确度(RE)在±7.8%以内。结论:该方法分析时间短、灵敏度高、专属性强,适用于伊曲康唑和羟基伊曲康唑的药动学研究。     

液相色谱-质谱联用法测定人血浆中多潘立酮

目的:建立测定人血浆中多潘立酮的液相色谱-电喷雾串联质谱(LC/ESI-MS/MS)法。方法:待测血浆0.1 mL 用甲醇沉淀蛋白,取离心后的上清液进样10μL在 Phenomenex Gimim-C_(18)柱(2.00 mm×50 mm,5 μm)上分离,流动相为甲醇-水(40:60,v/v,含0.3%醋酸),流速0.2 mL·min~(-1),LC/ESI-MS/MS 采用多离子反应监测,正离子模式,用于定量分析的离子反应分别为 m/z426→m/z175(多潘立酮)和 m/z 379→m/z 264(氨溴索,内标)。结果:血浆中的内源性物质不干扰测定,每个样品分析时间约2 min;本法线性范围为0.3～100 ng·mL~(-1),最低定量浓度为0.3 ng·mL~(-1);日内、日间 RSD 分别小于7.1%和13.3%,相对误差小于5.4%。结论:该法操作简便、快速、准确,灵敏度高,可用于多潘立酮临床治疗剂量的药物动力学研究。     

HPLC-MS/MS法测定人血浆中尼古丁浓度

目的建立简单、快速且灵敏的液质联用法检测人血浆中尼古丁的浓度。方法 血浆样品经二氯甲烷萃取后,采用Thermo Hypurity CN柱(2.1mm×150mm,5μm)分离,以乙腈-20mmol.L-1甲酸铵(50∶50,v/v)为流动相,流速为0.30mL·min-1。采用电喷雾离子源(ESI源)正离子多反应监测(MRM)扫描分析,尼古丁和d-4-尼古丁的离子选择通道分别为:m/z163→130和m/z 167→133.9。结果尼古丁的线性范围为0.597～76.48ng·mL-1,最低检测浓度为0.597ng·mL-1,平均提取回收率为70.7%～86.8%,日内和日间变异均<15%。结论本方法简单、灵敏、快速、重现性好,适用于尼古丁的人体临床药物代谢动力学及生物等效性研究。     

LC-MS/MS法测定人血浆中白藜芦醇的浓度

目的:建立液相色谱-串联质谱法测定人血浆中白藜芦醇浓度的方法。方法:血浆样品经四硼酸钠溶液碱化后,经甲基叔丁基醚提取处理。分析柱为RestekC1(82.1mm×150mm,5.0μm),保护柱为GminiC1(84mm×3.0mm,10.0μm),流动相为乙腈-水(85∶15,V/V,其中含0.2mmol·L-1乙酸铵),流速为0.3mL·min-1,进样量为5μL,内标为3,5-二羟基-4-异丙基二苯乙烯,使用电喷雾离子源,以负离子多重反应监测(MRM)方式进行检测。用于定量白藜芦醇和内标的MRM扫描离子通道分别为m/z227.0→143.0、253.2→211.0。每个样品的分析时间为5min。结果:白藜芦醇和内标的保留时间分别为1.43、2.55min。血浆中内源性物质对测定无干扰,白藜芦醇检测浓度在0.1～50.0ng·mL-1范围内同其与峰面积之比呈良好线性关系(r=0.9969),定量下限为0.1ng·mL-1。日内、日间RSD均低于11.25%,方法回收率为(91.70±3.81)%～(99.00±3.29)%。结论:本方法特异性强,灵敏度高,测定结果可靠,适用于白藜芦醇的血药浓度测定。     

高灵敏度液相色谱-串联质谱法测定人血浆中普拉克索

目的:建立灵敏、易操作的液相色谱(串联质谱法测定人血浆中普拉克索,并应用于临床药动学研究。方法:血浆样品经乙醚-二氯甲烷(3:2,v/v)液-液萃取后,以乙腈-10 mmol.L-1醋酸铵-甲酸(60:40:0.1,v/v/v)为流动相,Venusil ASB-C18柱(150 mm×4.6 mm,5μm)进行分离,采用电喷雾电离源,以选择反应监测(SRM)方式进行正离子检测,用于定量分析的离子反应分别为m/z212→m/z(111+126+153)(普拉克索)和m/z243→m/z210(石杉碱甲,内标)。结果:测定血浆中普拉克索的线性范围为5.92～740 pg.mL-1,定量下限为5.92 pg.mL-1,日内、日间精密度(RSD)均小于9.7%,准确度(RE)在-2.5%～0.6%之间。本法被成功应用于健康受试者单剂量口服0.125 mg和0.25 mg盐酸普拉克索片的药动学研究。结论:本方法同时选择3个主要碎片离子作为普拉克索定量产物离子,明显改进了分析方法的灵敏度,适用于人血浆样品中普拉克索的测定。     

LC-MS/MS法测定人血浆中替利定和去甲替利定的质量浓度

目的:建立LC-MS/MS法测定人血浆中替利定和去甲替利定的质量浓度。方法:选用Agilent-Zorbax-Eclipse-XDB-C18色谱柱,以甲醇-1 mmol·L-1醋酸铵(75∶25)为流动相,采用正离子,多反应监测方式测定样品质量浓度。用于定量分析的离子对分别为[M+H]+m/z 274.3→m/z 155.1(替利定),[M+H]+m/z 260.2→m/z 155.1(去甲替利定)和[M+H]+m/z 284.8→m/z 192.9(地西泮)。结果:血浆样品中,替利定在0.5～250 ng·mL-1范围内线性关系良好(r=0.9936),最低定量质量浓度为0.5 ng·mL-1;去甲替利定在1～500 ng·mL-1范围线性关系良好(r=0.9948),最低定量质量浓度为1 ng·mL-1。二者日内与日间RSD均小于15%,平均回收率高,且稳定性均较好。结论:本方法简便快速、灵敏准确、特异性强,适用于盐酸替利定和去甲替利定的体内药代动力学研究。     

液相色谱-质谱／质谱联用法测定人血浆中加兰他敏

目的:建立测定人血浆中加兰他敏浓度的液相色谱-质谱/质谱联用法,用于其人体血药浓度测定。方法:血浆样品经液-液萃取后,以甲醇-水-甲酸(65:35:1)为流动相,Zorbax SB-C_8柱(150mm×4.6mm,5μm)分离,采用大气压化学电离源,以选择反应监测方式进行正离子检测。内标为双氢吗啡酮,用于定量分析的离子反应分别为 m/z 288→m/z 213(加兰他敏)和 m/z 286→m/z 185(内标)。结果:血浆中加兰他敏最低定量限为0.5ng·mL~(-1),其线性范围为0.5～100ng·mL~(-1)。其高、中、低3个浓度的平均提取回收率为80.9%,方法回收率为100.5%,日内及日间 RSD 均<8%。结论:方法选择性强,灵敏度高,快速准确,可作为加兰他敏人体内药动学研究的手段。     

HPLC-MS/MS法测定人血浆中奥昔布宁和去乙基奥昔布宁的质量浓度

目的 建立HPLC-MS/MS法测定人血浆中奥昔布宁和去乙基奥昔布宁的质量浓度.方法 选用Waters-Atlantis dC18色谱柱,以0.01％甲酸溶液-乙腈为流动相进行梯度洗脱,采用正离子多反应监测方式测定样品质量浓度.用于定量分析的离子对分别为m／z358.2→142.2(奥昔布宁),m/z330.2→96.2(去乙基奥昔布宁),m/z268.2→152.2(奥昔布宁D10),m/z 335.2→101.2(去乙基奥昔布宁D5).结果 血浆样品中,奥昔布宁在0.05～25 ng·mL-1线性关系良好(r=0.996 5),最低定量浓度为0.05 ng·mL-1;去乙基奥昔布宁在0.05～25 ng·mL-1线性关系良好(r=0.998 5),最低定量浓度为0.05 ng· mL-1.两者日内与日间RSD均＜15％,提取回收率＞75％,且稳定性均较好.结论 本方法简便快速、灵敏准确、特异性强,适用于奥昔布宁和去乙基奥昔布宁的体内药物动力学研究.     

HPLC-MS/MS测定人体血浆和尿液中罗通定的浓度

目的 建立一种简便、灵敏的测定人体血浆和尿液中罗通定浓度的高效液相色谱串联质谱( HPLC-MS/MS)方法.方法 血浆、尿液样品分别采用乙腈沉淀处理后,进样分析.采用Agilent-ECLIPSE-C18柱(2.1mm×150 mm,5μm),以乙腈-0.2％甲酸溶液=85∶15为流动相,采用正离子,多反应监测方式测定样品浓度.检测离子为m/z356.3→192.3(罗通定)和m/z 285.0→193.0(地西泮,内标).结果 罗通定血浆及尿样均在2.5～1000 ng· mL-1与峰面积线性关系良好(r=0.999 4);最低定量浓度均为2.5 ng·mL-1.日内与日间RSD均＜10％,血浆样品回收率在91.4％～109.2％,尿样回收率在88.63％～115.8％.结论 本方法简便快速、灵敏准确,适用于罗通定在人体内的药物动力学研究.     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.