高效毛细管电泳法分析血样尿样中巴比妥类药物

建立了血、尿样品中巴比妥类药物的高效毛细管电泳分离测定方法，测定条件为：运行缓冲液配比：１００ｍｍｏｌ／Ｌ十二烷基硫酸钠－１００ｍｍｏｌ／Ｌ磷酸二氢钠－甲醇－水＝７０∶１５∶５∶１０（ｐＨ值调至７．５５±０．０５），操作电压１７ｋＶ，检测波长２００和２８５ｎｍ，２０ｍｉｎ内７种巴比妥类药物全部得到分离。考察并选择了体液样品的预处理方法，测定了血浆和尿液中６种巴比妥类药物浓度（苯巴比妥、甲基苯巴比妥、异戊巴比妥、硫喷妥、戊巴比妥和速可眠）。测定血药浓度的线性范围为５．０～３５μｇ／ｍｌ，尿样药物浓度线性范围为１．０～８．０μｇ／ｍｌ，最低检测浓度为１．０μｇ／ｍｌ，方法重现性为ＲＳＤ小于１３％。     

替硝唑片剂的HPLC和UV测定

采用ＨＰＬＣ和ＵＶ法测定替硝唑片剂的含量，其线性范围和平均回收率分别为：ＨＰＬＣ法６０～１００．μｇ／ｍｌ，９９．５６％（ＲＳＤ０．８８％）；ＵＶ法５～２５μｇ／ｍｌ，９９．７０％（ＲＳＤ０．５４％）。方法准确可靠。     

反相高效液相色谱法测定格列甲嗪血浆浓度

应用反相高效液相色谱法测定格列甲嗪血浆浓度。色谱柱为ＯＤＳＣ１８柱，流动相为乙腈─甲醇─水（４０：２０：４０Ｖ／Ｖ），内含０．１％冰醋酸和０．０８％三乙胺。流速０．５ｍｌ／ｍｉｎ，柱温５０℃，检测波长２７５ｎｍ。线性范围在０．０２～１．００μｇ／ｍｌ（ｒ＝０．９９９９）。检测下限为０．０１μｇ／ｍｌ。平均回收率为９９．０±２．８％，ＲＳＤ＝２．８２％。日内及日间测定相对标准偏差均小于５％。     

高效液相色谱法测定人血清中5-氟尿嘧啶的浓度

目的：测定 ５－氟尿嘧啶在人血清中的浓度。方法：三氯乙酸沉淀血清中蛋白质,高效液相色谱法测定含量。色谱柱为 ＳｈｉｍｐａｃｋＣＬＣ Ｃ１８柱,加 ＹＷＧ预柱;流动相为 ０．２５％磷酸二氢钾－乙腈（９８：２）,ｐＨ７．０;流速为 １ｍｌ／ｍｉｎ;紫外检测波长 ２６５ｎｍ。分别在低浓度范围（０．１９５～６．２５０μｇ／ｍｌ）和高浓度范围（６．２５０～２００．００μｇ／ｍｌ）制作标准曲线以用于定量。结果：５－氟尿嘧啶在 ０．１９５～２００μｇ／ｍｌ浓度范围呈良好的线性关系,０．８、４、２０和１００μｇ／ｍｌ４个浓度点的日内平均回收率和ＲＳＤ分别为１０４．８８％、１．９５％,１０４．５８％、１．３８％,１０１．４０％、０．３９％,９９．１４％、０．３７％;日间平均回收率和ＲＳＤ分别为１０５．１２％、２．０２％,１０６．３０％、０．７８％,１００．６０％、０．６５％,９９．３８％、０ ９２％。结论：本文建立的方法快速、准确,适合于５－氟尿嘧啶的药代动力学研究和常规血药浓度监测。     

高效液相色谱法同时测定人血清中茶碱与苯巴比妥、苯妥英、卡马西平的含量

本文报道茶碱与苯巴比妥（ＰＢ）、苯妥英（ＤＰＨ）、卡马西平（ＣＢＺ）两类（４种）不同药物血浓度的ＨＰＬＣ同时测定法．采用国产色谱柱ＹＷＧＣ１８（４．６×２５０ｍｍ），检测波长为２５４ｎｍ，流动相为甲醇－水（５０：５０，ｖ／ｖ），流速：１ｍｌ／ｍｉｎ，以４－氨基安替匹林作内标，各药物的平均回收率分别为茶碱９９．０２％，ＰＢ１００．７３％，ＤＰＨ１０１．０％，ＣＢＺ１０１．７７％。对各药血清标准液的峰高比测量的变异系数（ｎ＝１０），分别为茶碱（１０μｇ／ｍｌ）６．３％，ＰＢ（２０μｇ／ｍｌ）６．１％，ＤＰＨ（２０μｇ／ｍｌ）４．０％，ＣＢＺ（１０μｇ／ｍｌ）５．２％。本法具有快速、准确、适用性广的特点，用于治疗药物的监测，效果满意，大大提高了实验室的工作效率。     

零交导数分光光度法测定输液中茶碱和头孢哌酮钠含量

本文用零交导数分光光度法测定了输液中茶碱和头孢哌酮钠的含量，样品不经分离即可直接同时测定，茶碱在２～１０μｇ／ｍｌ与其在２２６．１ｎｍ（头孢哌酮钠的一阶导数零交点）处相应的振幅值呈良好的线性关系（γ＝０．９９９９，ｎ＝５）；而头孢哌酮钠在４～２４μｇ／ｍｌ范围与其在２４０．２ｎｍ（茶碱的一阶导数零交点）处相应的振幅值呈良好的线性关系（γ＝０．９９９９，ｎ＝６）。平均回收率为茶碱９９．９％＋０．５％；头孢哌酮钠９９．１％±０．７％。相对标准偏差分别为０．５％和０．７％。     

高效液相色谱法测定替硝唑片的含量

应用高效液相色谱法外标法测定替硝唑片剂的含量。色谱条件为色谱柱：Ｓｋｉｍ－ｐａｃｋＣＬＣ－ＯＤＳ１５０×６．０；检测波长：３１０ｎｍ；流动相：水－甲醇－冰醋酸（８０∶２０∶０．１）；流速：１ｍｌ／ｍｉｎ；灵敏度：０．０１ＡＵＦＳ；进样量：２０μｌ。其浓度在６０～１００μｇ／ｍｌ内与峰面积呈良好的线性关系，相关系数为０．９９９５。其精密度与稳定性的相对标准偏差分别（ＲＳＤ）为０．５％和０．８５％。平均回收率为９９．５６％，ＲＳＤ为０．８８％，结果准确可靠     

高效液相色谱法测定生物样品中两性霉素B

两性霉素Ｂ（ＡＭＢ）常用于内脏或全身真菌感染的治疗。由于ＡＭＢ对肝、肾等有较大毒性，应监测血浆中ＡＭＢ的浓度，以便调整其用量。我们采用高效液相色谱法（ＨＰＬＣ），在μ－ＢｏｎｄａｐａｋＣ１８柱（３．９ｍｍ×３００ｍｍ，１０μｍ）上，以０．０５ｍｏｌ／ＬＥＤＴＡ－２Ｎａ溶液－乙腈（１∶１）为流动相；流速为１．４ｍｌ／ｍｉｎ；检测波长为４０５ｎｍ，测定了血浆、脑脊液中ＡＭＢ浓度。ＡＭＢ血浆中最小检出量为０．０２μｇ／ｍｌ。血浆中ＡＭＢ提取率＞８７％。日内精密度在５．０１％～６．２８％之间，日间精密度血样＜７．８６％，ＣＳＦ＜５．９８％。血浆中ＡＭＢ浓度在０．０５～２．０μｇ／ｍｌ范围内有良好线性关系。方法回收率为９９．０４％±３．９０％。该法用于１例曲霉菌全身感染者ＡＭＢ血药浓度测定，为制定给药方案提供了依据。     

高效液相色谱法测定人血清中氯林可霉素

本文研究建立了人血清中氯林可霉素的高效液相色谱测定法（ＨＰＬＣ），该方法在超低紫外（１９８ｎｍ）下测定，具有取样量小（２００μｇ），线性范围宽（０．５％～５０μｇ／ｍｌ）；灵敏度高，以３倍基线噪音计算，最低检测限可达０．０５μｇ／ｍｌ；重现性好，高中低３个质控样品批间ＲＳＤ％分别为１．９７、０．４０和２．１５，批内ＲＳＤ％分别为０．８０、０．４６和２．３３，平均回收率分别为１０１．８７，１００．３４％和９３．８１％；方法操作简便等特点，为测定血清中药物浓度，确定给药方案和治疗剂量，以及药物动力学分析和药效学评价提供了方法学基础     

反相高效液相色谱法同时测定苯巴比妥和卡马西平血药浓度

目的：建立同时测定微量血浆中苯巴比妥和卡马西平浓度的方法。方法：采用ＲＰ－ＨＰＬＣ法,以艾司唑仑为内标,同时测定微量血浆中苯芭巴比妥和卡马西平浓度。色谱柱ｓｈｉｍａｄｚｕ ｓｈｉｒｍｐａｃｋ ＣＬＣ－Ｃ１８不锈钢柱,流动相为醇－水（６０：４０）,流速０．８ｍｌ．ｍｉｎ＾－１,检测波长２５４ｎｍ。结果：苯巴比妥在４～６０μｇ．ｍｌ＾－１浓度范围内线性良好（ｒ＝０．９９９８）,卡马西平在２～１６μｇ．ｍｌ＾－１浓度范围内线性良好（ｒ＝０．９９９５）,最低检测限分别为１１．５７ｎｇ．ｍｌ＾－１和４．９２ｎｍ．ｍｌ＾－１,两者高、中、低３种浓度的平均回收率分别为９９．９２％,１０１．３０％,９７．９２％和９９．４１％,１０１．５２％,９８．２２％（ｎ＝９）,日内ＲＳＤ分别为３．１％,２．６％,３．８％和１．９％,１．６     

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.

---

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.