高效毛细管电泳法测定白术中白术内酯Ⅰ的含量

摘 要 目的： 建立高效毛细管电泳( HPCE) 测定白术中白术内酯Ⅰ（Atractylenolide-1）含量的方法。方法: 运用高效毛细管电泳方法,熔融石英毛细管(75μmID ×50 cm) ,缓冲液为50 mmol·L^-1硼砂缓冲液,检测波长:210 nm ,分离电压:20 kV,柱温25℃,用0.45 μm微孔滤膜过滤后进样,压力进样:50 mbar×5 sec。结果: 方法最低检测浓度为0.5 μg·mL^-1,线性范围2～100 μg·mL^-1,r=0.996,线性关系良好。日内RSD为1.3% ,日间RSD为2.5%,平均回收率分别为97.1%。结论:本法简单、灵敏经济,可作为白术中白术内酯Ⅰ的含量的测定。     

高效毛细管电泳法测定地榆中没食子酸的含量

摘 要 目的： 建立高效毛细管电泳测定地榆中没食子酸含量的方法。方法: 优化出毛细管电泳的最佳测定条件及参数：PDA检测器,检测波长为220 nm;电泳缓冲液体系为20 mmol·L^-1的硼砂溶液（pH 9.0）;运行电压为15 kV;选取苯甲酸钠为内标物定量。结果: 没食子酸在0.200～6.000 μg·mL^-1范围内线性良好(r=0.999 6) ,LOD为0.050 μg·mL^-1,精密度RSD≤1.72 %,地榆的平均加样回收率为98.78%（RSD=2.57%,n=3）;测得地榆中没食子酸含量为0.31%。结论: 该方法准确,可靠,重复性好,可用于地榆中没食子酸含量的测定。     

高效毛细管电泳法同时测定维拉帕米和去甲维拉帕米对映体的血浆浓度

用高效毛细管电泳中最常用的毛细管区带电泳模式,以三甲基-β-环糊精(TM-β-CD)为手性选择剂,同时测定维拉帕米和去甲维拉帕米对映体的血浆浓度。手性测定条件:手性拆分缓冲液为含60mmol·L^-1TM-β-CD(pH2.5),浓度60mmol·L^-1的磷酸盐缓冲液;毛细管柱为30cm×75μm(ID)的聚丙烯酰胺涂渍柱;选择电迁移方式进样,电压12kV,时间7s;运行电压14kV;紫外检测波长200nm;设柱温为20℃;12min内完成检测。各对映体检测的线性范围为2.50ng·ml^-1～300.0ng·ml^-1,准确度和精密度达到生物样品分析的要求。用本法测定健康志愿者口服和静滴VPM后各对映体的血浆浓度,结果良好。     

毛细管电泳法测定复方磷酸可待因口服制剂的含量

摘 要 目的： 建立毛细管电泳法（CE）法同时测定复方磷酸可待因口服制剂中磷酸可待因、盐酸麻黄碱、马来酸溴苯那敏、马来酸氯苯那敏、愈创甘油醚等有效成分的含量。 方法: 色谱柱为贝克曼eCAP^TM未涂层石英毛细管柱（50 cm×75 μm）;分离电压为10 kV;缓冲溶液为20 mmol·L^-1磷酸盐缓冲液（pH7.5）;检测波长为214 nm。 结果: 各测定组分在考察浓度范围内线性关系良好（r≥0.995）,回收率均≥96%。 结论: 该方法可用于不同复方磷酸可待因口服制剂中有效成分的含量测定,简单有效,具有通用性,便于快速有效的市场监督。     

氧氟沙星对映体的毛细管电泳手性分离

用高效毛细管电泳和二极管阵列检测器,以环糊精为拆分试剂对氧氟沙星进行手性分离(最大分离度可达2.18)。发现衍生化环糊精有较好的分离效果,用含DM-β-CD40mmol·L^-1,KH₂PO₄70mmol·L^-1,pH2.5的缓冲液,在25℃电泳,20kV分离电压下得到优化的分离结果。     

毛细管区带电泳法测定重组人干扰素α-1b滴眼液中间甲酚含量

摘 要 目的：运用毛细管区带电泳法（CZE）检测重组人干扰素α 1b（rhIFNα 1b）滴眼液中间甲酚含量。方法: 优化的CZE方法参数为采用无涂层的熔融石英毛细管（60 cm×50 μm）,电泳缓冲液为50 mmol·L-1磷酸盐缓冲液（pH 8.0）,压力进样（1.0 psi）,分离电压25 kV,检测波长195 nm,毛细管温度25℃。结果: 间甲酚在0.05～0.8 mg·ml-1浓度范围内与峰面积呈良好线性关系（r=0.997 3）,最低检出限0.002 mg·ml-1。3批rhIFNα1b 滴眼液中间甲酚的相对含量分别为1.94,1.89和1.97 mg·ml-1,平均回收率分别为94.3%（RSD=1.57%,n=9）。结果与HPLC方法结果基本一致。结论: 新建毛细管区带电泳法简便,分析速度快,试剂消耗量小,结果准确,可以用于重组人干扰素α1b滴眼液中间甲酚含量的检测。     

HPLC法测定硫酸阿托品滴眼液中硫酸阿托品的含量

摘 要 目的： 建立测定0.05%硫酸阿托品滴眼液中硫酸阿托品含量的方法。方法: 色谱柱为Agilent C₁₈柱(250 mm×4.6 mm,5 μm),柱温为30 ℃,流动相A为磷酸盐缓冲液(0.05mol·L^-1磷酸二氢钠溶液,用稀磷酸或氢氧化钠调节pH至（3.0±0.1),流动相B为乙腈,A和B以一定的比例梯度洗脱,检测波长为210 nm,流速为1.0 ml·min^-1,进样量为10 μl。结果: 硫酸阿托品在21.500～430.000 μg·mL^-1范围内线性良好(r=0.999 8),定量限为38.74 ng,平均回收率为98.95 %,RSD为0.67%（n=9）。结论: 建立的方法简便、灵敏、专属性好、结果准确,适用于硫酸阿托品滴眼液中硫酸阿托品含量的测定。     

柱前衍生化RP-HPLC拆分酮洛芬对映异构体的研究

摘 要 目的：建立酮洛芬对映异构体的拆分方法。方法: 采用柱前衍生化RP HPLC法,以L-丙氨酸-β-萘胺（L-Ala-β-NA）为手性衍生化试剂,色谱柱：Hypersil ODS-2柱（250 mm×4.6 mm,5 μm）,流动相：乙腈-0.025 mol·L^-1磷酸二氢钾溶液（40∶60,v/v）,流速：1.0 ml·min^-1,检测波长：245 nm,柱温：25℃,进样量：10μl。结果: 酮洛芬对映体衍生物获得较好的基线分离,S-(+) 对映体和R-(-) 对映体衍生物的色谱峰保留时间分别在24.2 min和26.0 min处。右旋酮洛芬在0.025~0.125 mg质量范围内线性关系良好（r=0.998 1）,平均回收率为90.93%（RSD=4.10%,n=9）。结论:该方法简便、准确、快速,为酮洛芬的光学异构体的测定提供了参考依据。     

苯磺酸左旋氨氯地平的毛细管电泳手性分离与纯度检查

目的：建立手性分离方法并检查苯磺酸左旋氨氯地平的光学纯度。方法：通过手性拆分剂的种类及浓度、缓冲液的pH及浓度、温度及电压的优化，选择最佳的手性分离条件。结果：最佳分离条件缓冲液为50mmol·L^-1，Tris（用磷酸调节pH2.5，含10mmol·L^-1羧甲基-β-环糊精）；检测波长为214nm;电压为25kV；温度为22℃，基线分离氨氯地平2个对映体，对建立的方法进行方法学验证，并检查实际样品的光学纯度。结论：建立的毛细管电泳法实用性强，费用低，因此采用了高效毛细管电泳法测定光学纯度，为该药的质量控制和稳定性研究提供了可靠的分析方法。     

毛细管电泳手性拆分西他沙星异构体

目的:建立西他沙星及其异构体高效毛细管电泳拆分的新方法。方法:以β-环糊精及几种环糊精衍生物作为手性添加剂,采用毛细管电泳分离新型抗菌素西他沙星与其3个异构体。对运行缓冲液的浓度、pH,环糊精的种类、二元环糊精体系中环糊精的比例,分离电压,柱温,有机改性剂等因素对分离的影响进行考察,并对拆分机制进行初步探讨。结果:运行缓冲液为含2%硫酸化-β-环糊精和1%β-环糊精的25 mmol.L-1的磷酸二氢钾-磷酸缓冲溶液(pH 2.5),操作电压为-25kV,柱温为25℃,检测波长为295 nm,西他沙星与其3个异构体达到良好分离。结论:所建立的毛细管电泳方法简便,适用于西他沙星异构体的检查。     

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.