阳离子色谱柱-HPLC法测定人血浆中氯氮平及去甲氯氮平浓度

目的:建立同时测定人血浆中氯氮平及去甲氯氮平浓度的阳离子色谱柱-HPLC法。方法:用强阳离子交换色谱柱-HPLC测定人血浆中氯氮平及去甲氯氮平浓度,色谱柱为强阳离子交换色谱柱(4.6 mm×100 mm,5μm),流动相为乙腈-0.01 mol·L-1磷酸铵溶液(pH 5.1)(60∶40),流速为1.2 mL·min-1,紫外检测波长为257 nm,以AF2672为内标,含氯氮平及去甲氯氮平的血样经C8固相小柱萃取后进样。结果:阳离子色谱柱-HPLC测得氯氮平、去甲氯氮平的色谱峰峰形对称、无内外源性物质干扰,检测结果稳定、可靠、重复性好;通过改变流动相pH易于调整各化合物保留时间,同时去除空白血浆干扰;固相萃取前处理简单、省时且萃取回收率较高。氯氮平及去甲氯氮平浓度在50.4～990 ng·mL-1范围内线性关系良好;氯氮平及去甲氯氮平低(50.4 ng·mL-1)、中(396 ng·mL-1)、高(990 ng·mL-1)3个浓度的萃取回收率均大于91%,日内和日间RSD均小于4%(n=5)。结论:阳离子色谱柱-HPLC法测定人血浆中氯氮平及去甲氯氮平浓度,色谱峰峰形对称,保留时间适宜且无杂质干扰,适用于临床氯氮平及去甲氯氮平血药浓度监测和药动学研究。     

固相萃取-HPLC法快速测定苯妥英钠及卡马西平的血药浓度

目的 应用固相萃取(SPE)-反相HPLC法同时测定苯妥英钠(PHT)及卡马西平(CBZ)血药浓度。方法 将1ml血浆样品在GDX-403固相萃取柱上进行固相萃取，洗脱液混匀后直接进样。色谱条件：色谱：Hypersil BDS C18柱，流动相：甲醇：水=48：52，检测波长：235nm，流速：1ml／min，柱温：25℃，内标：非那西丁。结果 PHT在4．10-41．0μg／ml范围内线性良好(γ=0．9999)；CBZ在2．50-24．96μg／ml范围内线性良好（γ=0.9998)，方法平均回收率各为101.77％和101．87％，日内及日间误差分别小于4．13%和49.8％，最低检测浓度为：1．025μg／ml和0．156μg/ml。结论 该方法、准确、快速，可用于PHT及CBZ血浆浓度的同时测定。     

HPLC法测定复方冲洗剂中盐酸小檗碱的含量

目的建立高效液相色谱测定复方冲洗剂中盐酸小檗碱含量的方法。方法Discovery C18色谱柱，流动相：乙腈-0．0．5moL／L磷酸二氢钾(10％氢氧化钾调节pH为5．0)(30：70)，检测波长：350nm。结果线性关系：Y=0．294X-0．864，相关系数r=0．9997，线性范围在20.45～245．4ng，回收率：平均100．1％(n=5)，RSD=0．8％。结论本法操作简便准确，可作为复方冲洗荆中盐酸小檗碱含量的质量控制方法。     

高效液相色谱-质谱联用法测定人血浆中奥昔布宁的浓度

目的 建立一种用高效液相色谱-电喷雾离子化质谱联用技术测定奥昔布宁血药浓度的方法。方法 以0．0lmol／L乙酸铵水溶液-甲醇(15：85)为流动相，盐酸非洛普为内标，血浆样品经用环己烷萃取后上样，经C18柱分离后，以质谱为检测器，采用选择性离子检测(SIM)测定人体血浆中奥昔布宁的浓度。结果 线性范围0．2～50ng／mL(r=0．9948)，平均相对回收率在90％～110％之间，日内和日间精密度的RSD均小于10％，奥昔布宁的定量限为0．2ng／mL，提取回收率大于90％。结论 该方法快速、准确、灵敏，可用于奥昔布宁的药代动力学研究。     

盐酸二甲双胍缓释片的HPLC测定

建立了RP—HPLC法测定盐酸二甲双肌缓释片的含量及有关物质。采用ODS C18色谱柱，流动相为甲醇—5mmol/L磷酸二氢铵水溶液(用磷酸调pH至3．5，含0．1％SDS)(65：35)，检测波长为232nm。线性范围0．005～1．0mg／m1，r＝0．9999；检测限0．1ng。平均回收率为99．7％，RSD为0．5％。     

GC—MS定性定量分析生物检材中的安眠酮

目的：建立定性定量检测人体尿液、血液及唾液中安眠酮的分析方法。方法：尿液、血液、唾液等生物样本中的安眠酮经固相萃取法进行提取后,用气相色谱-质谱仪进行定性定量分析,从线性、抗干扰性、精密度、回收率及稳定性等方面对方法进行验证。结果：安眠酮浓度在0—400．0ng·mL^-1的范围内线性良好（r≥0．999）,方法的最低检出量为8．0ng·mL^-1;最低定量限为100．0ng·mL^-1;方法的日内、日间精密度CV≤5％（n=6）;各基质中前处理回收率在±10．0％（n=3）范围内。结论：本方法检测准确、灵敏、特异性好,可满足尿液、血液、唾液等生物检材中安眠酮的定性定量分析。     

高效液相色谱荧光检测法测定人血浆中佐匹克隆浓度

目的：建立高效液相色谱荧光法测定人血浆中佐匹克隆浓度的方法。方法：碱化血浆后经无水乙醚提取，挥干乙醚后，残渣经溶有参照标准品氢氯噻嗪的流动相溶解后进行测定。荧光检测的激发波长为315nm,发射波长为380nm，结果：本法测定佐匹克隆的线性范围为1－50mg.L^-1(r=0.999)，最低检测限0．2ng，最低检测浓度为1mg.L^-1，低浓度（1mg.L^-1)的日内变异系数为9．6％，日间变异系数为12．4％，低浓度萃取回收率和方法回收率分别是54．7％和87．0％，中高浓度的萃取和方法回收率均＞90％，结论：用外标法测定人血浆中佐匹克隆浓度，方法精密，准确、稳定，适用于佐匹克隆的临床检测和药代动力学研究。     

毛细管色谱法测定洛美沙星中的有机溶剂残留量

目的：建立毛细管气相色谱法测定洛美沙星中的有机溶剂残留量。方法：用INNOWAX毛细管气相色谱柱，FID检测器，以2-戊酮为内标进行测定。结果：乙酸乙酯、四氢呋喃、乙醇、乙腈的线性范围分别为0～80μg／ml(r=0．9997)、0～11．52μg／ml(r=0．9996)、0～80μg／ml(r=0．9997)，0～6．56μg／ml(r=0．9996)；平均回收率分别为100．5％、100．1％、101．2％、100．1％；RSD分别为1．30％、0．9％、1．18％和1．23％(n=9)。结论：本方法简单、准确、灵敏度高、重现性好，适用于洛美沙星中有机溶剂残留量的测定。     

RP-HPLC法测定硝酸咪康唑乳膏含量内外标比较

本文采用反相高效液相色谱内外标两种方法测定硝酸咪康唑乳膏的含量。固定相：ODS-C18；流动相：0．5％醋酸铵溶液-乙腈-甲醇（15；42．5；42．5）；流速：1．0ml·min^-1；紫外检测波长：230nm；内标法回收率100．44％，外标法回收率102．27％；两种方法的日内、日间相对标准偏差均小于2％。硝酸咪康唑的最低检测限为3ng，定量限为6ng；线性范围：0．3848-1．924mg·mL^-1，内标法r=0．9999；外标法r=0．9995。     

高效液相色谱法测定养胃舒颗粒中橙皮苷的含量

目的建立一种高效液相色谱法测定养胃舒颗粒中橙皮苷的含量。方法以ODS—C18为固定相,乙腈-0．5％磷酸（18：82）为流动相,检测波长为284nm。结果橙皮苷线性范围为98．75ng～3160ng,r=0．9999,加样回收率为98．02％,RSD为0．52％。结论该方法简便快速、准确,适合养胃舒颗粒的质量控制。     

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.     

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.     

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.