HPLC法测定地西他滨中L-异构体和α-异构体含量

目的 建立高效液相法对地西他滨中L-异构体和α-异构体进行拆分并检测其中异构体含量.方法 色谱柱采用CHIRACPAK-ADH手性柱(250mm ×4.6mm,5μm);流动相:无水乙醇一正己烷(50∶50);流速:0.5mL·min-1;检测波长:244nm;选样量:20μL.结果 本法对L-异构体和α-异构体的分离度为2.42,地西他滨浓度在0.395～4.02μg·mL-1范围内呈良好的线性关系(r =0.9999),L-异构体浓度在0.419 ～4.04μg·mL-1范围内呈良好的线性关系(r=0.9997),α-异构体浓度在0.419 ～4.02μg·mL-1范围内呈良好的线性关系(r =0.9998),检测限分别为5.10ng和5.08ng,平均回收率分别为98.47％和99.22％.结论 该方法操作简便,分离效果好,可用于地西他滨中异构体的含量测定.     

毛细管电泳法同时测定元胡止痛片中欧前胡素和异欧前胡素的含量

目的:建立以毛细管电泳法同时测定元胡止痛片中欧前胡素和异欧前胡素含量的方法。方法:以盐酸小檗碱为内标,采用未涂层弹性融硅石英毛细管柱(55cm×75μmID,有效长度47cm),以20mmol·L-1磷酸二氢钠+100mmol·L-1十二烷基磺酸钠+80%甲酰胺溶液(pH7.12)为运行缓冲液,分离电压为15kV,重力进样5s(高度15cm),检测波长为254nm。结果:欧前胡素、异欧前胡素检测浓度分别在8.0～40.0μg·mL-(1r=0.9987)、4.0～20.0μg·mL-(1r=0.9990)范围内与各自峰面积积分值呈良好线性关系;二者平均回收率分别为101.3%和100.2%,RSD分别为3.24%和1.44%(n=6)。结论:本法简便、快速、准确,可用于元胡止痛片的质量控制。     

高效毛细管电泳法测定育亨宾树皮中育亨宾的含量

目的建立高效毛细管电泳(HPCE)测定育亨宾树皮中育亨宾含量的方法。方法运用高效毛细管电泳方法,熔融石英毛细管(75μmID×50cm),缓冲液为20mmol·L-1磷酸盐缓冲液(PH=3.0),检测波长:270nm,分离电压:15kV,柱温25℃,用0.45μm微孔滤膜过滤后进样,压力进样:50mbar×5s。结果方法最低检测浓度为0.1μg·mL-1,线性范围1～200μg·mL-1,r=0.9991,线性关系良好。精密度RSD为1.3%,回收率为98.4%。结论本法简单、灵敏经济,可作为育亨宾树皮中育亨宾的含量的测定。     

微乳液相色谱法同时测定大黄中5种蒽醌类衍生物的含量

目的:建立同时测定大黄中芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚含量的方法。方法:采用微乳液相色谱法。以微乳为流动相,通过对影响分离度和保留时间的因素进行考察,优化最佳微乳体系组成为2.5%(W/V)十二烷基硫酸钠-0.1%(V/V)正辛烷-8.0%(V/V)正丁醇-0.5%(V/V)三乙胺(磷酸调pH值至3.0);色谱柱为HypersilODS2(250mm×4.6mm,5μm),柱温为28℃,流速为1mL·min-1,检测波长为254nm。结果:芦荟大黄素、大黄酸、大黄素、大黄酚和大黄素甲醚的进样浓度线性范围分别为4.7～47.0μg·mL-1(r=0.9999)、5.1～51.0μg·mL-1(r=0.9996)、5.5～55.0μg·mL-1(r=0.9996)、6.3～63.0μg·mL-1(r=0.9999)和0.4～40.0μg·mL-1(r=0.9987);五者平均加样回收率分别为98.67%、97.53%、101.37%、99.34%和99.52%,RSD(n=6)分别为0.31%、0.38%、0.49%、0.53%和0.87%。结论:本方法简便、准确,可用于大黄中5种蒽醌类衍生物的含量测定。     

顶空气相色谱法限量检测紫杉醇中残留的甲醇、丙酮和正己烷

目的建立紫杉醇中残留的甲醇、丙酮和正己烷的限量检测方法。方法DB-624毛细管柱(30m×0.53mm×3.0μm);氮气为载气,流速2.0mL·min-1;柱温:60℃保持15分钟,以30℃·min-1的速率升温至180℃,保持5分钟;进样口温度:180℃,分流比5:1;检测器:FID,200℃。顶空进样器:顶空瓶温度85℃,平衡时间30分钟;传输管温度110℃;定量管温度105℃;加压时间0.5min;进样时间1min。结果甲醇、丙酮和正己烷的线性范围分别为:12.00～180.1μg·mL-1(r=0.9996)、1.996～299.5μg·mL-1(r=0.9999)和0.2314～17.36μg·mL-1(r=0.9995);最低检测限分别为2.4μg·mL-1、0.57μg·mL-1和0.048μg·mL-1。结论本方法适合于限量检测紫杉醇中残留的甲醇、丙酮和正己烷。     

槐角丸中染料木素的高效毛细管电泳测定

目的:建立适合槐角丸中染料木素测定的毛细管电泳方法。方法:采用高效毛细管电泳法。实验时,先分别用0.1 mol·L-1盐酸溶液、0.1 mol·L-1氢氧化钠溶液及运行缓冲液冲洗未涂层弹性石英毛细管(50μm ×43 cm)10 min,然后以槲皮素为内标,运行电压28 kV,运行缓冲液为20 mmo|·L-1磷酸盐缓冲液(pH 9.2),检测波长254 nm,室温下电动进样5 s。结果:染料木素的线性范围为6.25-31.26μg·mL-1(r=0.9994),其高、中、低3个量的加样回收率分别为100.7％,100.1％和99.2％,RSD分别为1.5％,0.63％和1.7％。结论:本法用于测定槐角丸中染料木素的含量快速、简便、准确可靠,有良好的重复性。     

RP-HPLC法测定人体血浆中吉西他滨的浓度

目的:建立一种测定人体血浆中吉西他滨血药浓度的高效液相色谱方法。方法:取血浆样品1 mL,加内标100μL(含氟脲苷0.8μg·mL-1),混匀,加甲醇-乙腈(1:9)3 mL混匀,放置5 min,离心(3500 r·min-1,10 min),取上清液于60℃水浴放置,氮气吹干,残渣用0.5 mL流动相溶解,离心(15000 r·min～1,10 min),取上清液,进样50μL。色谱柱:Lichrospher 5-C18(4.6 mm×250 mm,5 μm),流动相:40 mmol·L-1醋酸铵缓冲液(用醋酸调节pH=5.5)-乙腈(97.5:2.5),流速:0.8 mL·min～1,检测波长:268 nm,柱温:25℃。结果:本方法线性范围0.20—10.0μg·mL～1,r=0.9999,方法检测限为0.10μg·mL-1(S/N>4),定量限为(0.21±0.02)μg·mL-1(S/N>10);方法回收率为100.1％-106.6％(n=21),日内RSD为2.3％-4.0％(n=21),日间RSD为3.2％-5.2％(n=21)。结论:本方法灵敏度高,操作简便、准确,可用于人体血浆中吉西他滨浓度的测定及药动学研究。     

高效液相色谱法测定他达拉非片的有关物质

目的 采用高效液相色谱法测定他达拉非片的有关物质.方法 采用C8柱,以0.01％三氟乙酸溶液为流动相A,乙腈为流动相B,流速为1.0mL·min-1,梯度洗脱;检测波长285nm.结果 在该色谱条件下,杂质峰与主峰均能有效分离,杂质G与他达拉非的线性范围分别为0.016～1.614μg·mL-1(r=0.9994)及0.008～0.815μg·mL-1(r=0.9995),平均回收率分别为101.23％及102.65％.结论 本法简便、准确,专属性强,有助于更好地控制本品的质量.     

超高效液相色谱法测定富马酸伏诺拉生原料有关物质

目的：建立超高效液相色谱法测定富马酸伏诺拉生有关物质的方法。方法：采用ZORBAX SB-C18色谱柱(4.6 mm×150 mm,1.8 μm),流动相A为0.025 mol·L-1磷酸盐缓冲溶液(pH 6.0)- 甲醇-乙腈(14∶5∶1),流动相B为0.025 mol·L-1磷酸盐缓冲溶液(pH 6.0)-乙腈(3∶7), 采用梯度洗脱,流速1.0 mL·min-1,柱温30 ℃,检测波长230 nm,进样量20 μL。结果：杂质A、 杂质C、杂质D、杂质F、杂质G、杂质H、杂质J、杂质K、杂质L、杂质M、杂质N和伏诺拉生分别在0.04177~2.088 μg·mL-1(r=1.0000)、0.07479~3.740 μg·mL-1(r=1.0000)、0.07335~3.668 μg·mL-1(r=1.0000)、0.1000~2.004 μg·mL-1(r=1.0000)、0.1325~6.623 μg·mL-1(r=1.0000)、 0.09300~1.860 μg·mL-1(r=1.0000)、0.1380~2.7530 μg·mL-1(r=1.0000)、0.1350~2.698 μg·mL-1 (r=1.0000)、0.1130~2.257 μg·mL-1(r=1.0000)、0.05060~2.530 μg·mL-1(r=0.9999)、 0.1420~2.846 μg·mL-1(r=1.0000)、0.1190~2.382 μg·mL-1(r=0.9998)浓度范围内与峰面积呈良好线性关系。上述各杂质的平均回收率(n=9)分别为97.24%、102.12%、99.46%、98.29%、101.51%、 100.84%、99.18%、101.20%、98.49%、103.63%、96.49%。采用该检测方法对5批样品进行有关物质检测,其中杂质C、杂质M、杂质N检出量均≤0.05%,杂质H、其他最大单个杂质检出量均≤0.10%,总杂质检出量均≤0.31%。结论：经方法学验证,本法专属性强、灵敏度高、准确度高,可用于富马酸伏诺拉生原料的有关物质测定,为其质量控制提供方法保证。     

毛细管气相色谱法测定盐酸利多卡因注射液的有关物质

摘要：目的:建立毛细管气相色谱法测定盐酸利多卡因注射液有关物质的方法。方法:采用HP-1毛细管色谱柱（30 m×0.32 mm, 1.00μm）以气相色谱法测定盐酸利多卡因注射液的有关物质。起始温度120℃,以10℃·min-1升温速率升温至210℃,维持15 min,检测器为氢火焰离子化检测器。结果:GC系统中所有杂质均可完全分离。杂质2,6-二甲基苯胺、杂质B和杂质H的浓度分别在0.087 28～4.364μg·ml-1（r=0.999 9）、0.548～13.7μg·ml-1（r=0.999 8）、0.434～10.85μg·ml-1（r=0.999 5）范围内与峰面积成良好的线性关系。平均回收率分别为99.4%,99.1%,99.4%,RSD分别为1.4%,3.8%,4.4%（n=9）。3批样品中,2,6-二甲基苯均有检出,含量均为0.01%;杂质B、杂质H均未检出;其他单个杂质含量分别为0.3%,0.3%,0.2%。杂质总和分别为0.3%、0.3%、0.2%。结论:本方法准确可靠,可用于盐酸利多卡因注射液有关物质的测定。     

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.