液质联用测定人血浆中伪麻黄碱的浓度

目的:建立液质联用法测定人血浆中伪麻黄碱的浓度。方法:采集到的血浆样品以苦参碱为内标,经0.2 mL 0.02 mol·mL~(-1)碳酸钠溶液碱化后氯仿萃取进行 LC-MS 分析。色谱条件为 Kromasil C_(18)柱(150 mm×4.6 mm,5 μm),流动相为乙腈-0.1%甲酸(13:87),流速为1.0 mL·min~(-1);质谱条件为大气压电喷雾离子源(ESI 源),正离子方式检测;扫描方式为选择离子监测(SIM),用于定量分析的离子分别为 m/z 166(伪麻黄碱)和 m/z 249(苦参碱)。结果:方法的线性范围为5～300 ng·mL~(-1),定量下限为5 ng·mL~(-1),提取回收率均大于77.9%,日内、日间精密度(RSD)均小于12.7%。结论:本方法灵敏、专属,适于伪麻黄碱人体血浆药物浓度的测定。     

LC-MS/MS法同时测定大鼠血浆中甘草酸和甘草次酸

目的：建立一种同时测定大鼠血浆中甘草酸和其代谢产物甘草次酸的高效液相色谱一串联质谱的（LC-MS／MS）分析方法。方法：采用汉邦VP—ODS（150min×2．1nun,5μm）色谱柱,流动相为甲醇和1％的甲酸水,梯度洗脱：21％水相（0-3．5min）,2％水相（3．5-8．2min）。简单的乙腈沉淀蛋白法处理血浆样品。以格列喹酮为内标,采用ESI源负离子模式、多反应检测进行测定。检测离子对分别为m／z821．4→350．9（甘草酸）、m／z526→401．1（格列喹酮）和m／z469．2→425．2（甘草次酸）。结果：甘草酸和甘草次酸的检测浓度分别在50－2000ng·mL-1、10-500ng·mL-1范围内线性关系良好。相关系数为0.9989和0．9981。日内及日间精密度RSD均小于15％。结论：方法专属性强．准确度好．适用于同时监测甘草酸和甘草次酸的浓度及进行药动学研究。     

液相色谱-质谱联用法测定大鼠血浆中盐酸金刚烷胺浓度及体内药动学研究

目的:建立测定大鼠血浆中盐酸金刚烷胺的液相色谱-质谱联用法。方法:色谱柱为 Agilent Zorbax SB-C_(18)(2.1 mm×150 mm,3.5μm);乙腈-0.05%甲酸溶液(28∶72)为流动相,流速0.2 mL·min~(-1);柱温25℃;内标为盐酸美金刚。质谱条件为电喷雾电离源(ESI),选择正离子抽提方式检测,金刚烷胺和美金刚的选择检测离子分别为 m/z 152.20([M H]~ )、m/z180.20([M H]~ )。选用 SD 大鼠6只,单剂量尾静脉注射给予1 mg·kg~(-1),进行了血药浓度的测定。结果:盐酸金刚烷胺血浆线性范围为0.9824～491.2 ng·mL~(-1),检测限为0.025 ng·mL~(-1),方法回收率均大于95%,日内、日间精密度试验的 RSD<5.0%。结论:本方法专属性强,灵敏度高,线性关系良好,操作简便,适用于药代动力学研究。     

液相色谱-电喷雾质谱联用法测定人血浆中二甲双胍

目的:建立测定人血浆中二甲双胍浓度的液相色谱-电喷雾质谱联用(LC/ESI-MS-MS)法。方法:待测血浆0.2 mL经甲醇沉淀除去蛋白,离心,取上清液5 μL在氰基柱上分离,流动相为甲醇-20 mmol·L~(-1)醋酸铵水溶液(30:70),流速为1.0 mL·min~(-1),LC/ESI-MS-MS 多反应离子检测,正离子模式,用于定量分析的离子分别是二甲双胍 m/z 130.1→70.7和吗啉双胍(内标)m/z 172.2→59.6。结果:血浆中无干扰测定的内源性物质,每个样品分析时间小于5 min,线性范围为10.08～1008 ng·mL~(-1),定量下限为10.08 ng·mL~(-1),日内、日间精密度均小于10%,提取回收率大于85%。结论:该法操作快速、简单、准确、灵敏度高,适用于临床药代动力学研究。     

HPLC-MS／ESI法同时测定人血浆中盐酸特拉唑嗪盐酸哌唑嗪和甲磺酸多沙唑嗪

目的:建立同时测定血浆中盐酸特拉唑嗪、盐酸哌唑嗪和甲磺酸多沙唑嗪的 HPLC-MS/ESI 方法。方法:血浆样品0.5mL,加入10 ng·mL~(-1)二盐酸氟哌噻吨内标液50 μL,经饱和碳酸钠200 μL碱化和1.25 mL 正已烷-叔丁基甲醚(1:1)萃取;以Hypersil GOLD C_(18)柱(100 mm×2.1 mm,5 μm)为固定相,20 mmol·L~(-1)醋酸铵(0.05%甲酸,pH 4.2)-乙腈-甲醇为流动相,梯度洗脱分离,流速为0.2 mL·min~(-1);采用 HPLC-MS,选择离子检测(SIM)法测定盐酸特拉唑嗪([M H]~ ,m/z 388)、盐酸哌唑嗪([M H]~ ,m/z 384)、盐酸多沙唑嗪([M H]~ ,m/z 452和内标氟哌噻吨([M H]~ ,m/z 435)的血药浓度。结果:线性范围为1～100 ng·mL~(-1),r>0.9930(n=7);萃取回收率大于83.0%;方法回收率大于97.0%;日内、日间精密度 RSD 均小于10.0%。结论:该方法灵敏、专属、快速,适用于3种药物的血药浓度监测,以及药代动力学和生物等效性的研究。     

液相色谱-电喷雾串联质谱法测定人血浆中辛伐他汀浓度

目的:建立液相色谱-电喷雾串联质谱联用测定人血浆中辛伐他汀浓度的方法。方法:色谱条件为 Discovery ODS C_(18)(2.1 mm×100 mm,3μm)色谱柱;流动相为乙腈-0.1%甲酸溶液(65:35,v/v);柱温30℃;流速0.2 mL·min~(-1);进样量5μL。质谱条件为电喷雾电离源(ESI),选择性检测定量离子为 m/z 441.3/325.0(辛伐他汀)和 m/z 427.2/325.0(洛伐他汀,内标)带正电荷的离子峰。样品用乙酸乙酯提取。结果:血浆中辛伐他汀浓度在0.20～20.0 ng·mL~(-1)内呈良好线性关系,r=0.9996。提取回收率为90.0%～92.4%(RSD 6.0%～14.9%),方法回收率在85%～115%之内,日内和日间精密度试验的RSD 均<15%,最低检测浓度为0.2 ng·mL~(-1)。结论:该测定方法经全面考察符合血浆样品的测定要求,可以应用于辛伐他汀的人体药动学研究和生物等效性评价。     

HPLC—MS法同时测定人血浆中普伐他汀及其主要代谢物3’α-异普伐他汀

目的:建立同时测定人血浆中普伐他汀及其主要代谢物3’α-异普伐他汀的HPLC—MS法,并研究普伐他汀钠片在人体内的药代动力学。方法:血浆样品中加入内标霉酚酸,用固相萃取柱进行提取。色谱柱为Discovery C_(18)柱(5μm,150mm×4.6 mm),流动相为甲醇-乙腈-6 mmol·L~(-1)乙酸铵溶液(20:30:50),流速为0.4 mL·min~(-1)。采用HPLC—ESI~--MS法,选择离子检测方式,用于定量分析的检测离子为m/z423.4(普伐他汀和3’α-异普伐他汀)和m/z 319.2(内标)。结果:血浆中内源性物质对样品测定无干扰。普伐他汀和3’α-异普伐他汀的线性范围均为1.25—200 ng·mL~(-1),最低定量浓度均为1.25 ng·mL~(-1),提取回收率均大于80%,日内、日间RSD均小于10%。结论:本法灵敏、准确,适用于普伐他汀及其主要代谢物3’α-异普伐他汀的药代动力学研究。     

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。     

LC-MS／MS法测定人血浆中米格列醇

目的:建立测定人血浆中米格列醇的 LC-MS/MS 法。方法:以盐酸格拉司琼为内标,以 Kromasil CN 柱(2.1 mm×150mm,3.0μm)为分析柱;采用0.05%三氟乙酸乙腈溶液-0.05%三氟乙酸水溶液(80:20)为流动相;流速0.2 mL·min~(-1);柱温45℃。质谱条件为电喷雾电离源(ESI~ ),以选择反应离子监测(SRM)方式进行检测,用于定量分析的反应离子分别为 m/z208.1→146.1(米格列醇),m/z 313.1→138.0(盐酸格拉司琼)。结果:米格列醇在0.01～4.0μg·mL~(-1)浓度范围内线性良好;最低检测限为2.0 ng·mL~(-1)(S/N=4);日内、日间精密度(RSD)均小于7%;低、中、高3个浓度的方法回收率均大于90%。结论:本法专属性强,灵敏度高,样品处理简单,可用于米格列醇临床药物动力学研究。     

液相色谱-串联质谱法快速测定人血浆中伊曲康唑

目的:建立灵敏、快速的液相色谱-串联质谱法测定人血浆中伊曲康唑,并用于药代动力学研究。方法:血浆样品经乙腈沉淀蛋白后,以乙腈-水-甲酸(80:20:0.2,v/v/v)为流动相,流速0.50 mL·min~(-1),Zorbax sB C_(18)柱分离,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/z 705→m/z(392 432)(伊曲康唑)和m/z 256→m/z 167(内标,苯海拉明)。结果:测定血浆中伊曲康唑的线性范围为1.00～1000 ng·mL~(-1),定量下限为1.00 ng·mL~(-1)。日内、日间精密度(RSD)均小于7.2%,准确度(RE)在±3.0%以内。应用本法测得20名健康受试者单剂量口服200mg 伊曲康唑胶囊后的主要药动学参数为:T_(max)(4.25±1.02)h,C_(max)(155.5±73.3)ng·mL~(-1),t_(1/2)(20.4±11.4)h;用梯形法计算,AUC_(0-84h)(2257±1168)ng·h·mL~(-1),AUC_(0-∞)(237.1±1207)ng·h·mL~(-1)。结论:该法选择性强、灵敏度高、操作简便,适用于伊曲康唑的临床药代动力学研究。     

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.     

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.