RP-HPLC法同时测定不同产地栀子中栀子苷、西红花苷-Ⅰ和西红花苷-Ⅱ的含量

目的:建立同时测定栀子中栀子苷、西红花苷-Ⅰ和西红花苷-Ⅱ含量的方法,并对14批不同产地栀子药材中3种成分的含量进行测定。方法:采用反相高效液相色谱法。色谱柱为Thermo ODSC1(8250mm×4.6mm,5μm),流动相为甲醇-0.1%磷酸水溶液(梯度洗脱),流速为1.0mL·min-1,检测波长分别为栀子苷238nm、西红花苷-Ⅰ和西红花苷-Ⅱ440nm。结果:栀子苷、西红花苷-Ⅰ、西红花苷-Ⅱ的进样量分别在0.448～5.600、0.142～1.775、0.026～0.325μg范围内与各自峰面积积分值呈良好线性关系,r分别为0.9992、0.9998、0.9999;三者平均回收率分别为99.69%(RSD=2.44%,n=6)、98.26%(RSD=2.62%,n=6)、102.94%(RSD=3.13%,n=6)。结论:本方法简便、准确、重复性好,可同时测定栀子中栀子苷、西红花苷-Ⅰ和西红花苷-Ⅱ的含量。     

反相离子对高效液相色谱法测定盐酸黄酮哌酯片的含量

目的:建立一种采用反相离子对高效液相色谱法测定盐酸黄酮哌酯片含量的方法。方法:用Shim-Pack vp-ODS柱(4.6mm×150mm,5μm),以0.05mol·L~(-1)磷酸二氢钾(含1％三乙胺,用磷酸调pH3.8)-0.0025mol·L~(-1)庚烷磷酸钠甲醇溶液(1:1.2)为流动相,流速1mL·min~(-1),UV检测波长241nm。柱温:室温,进样量20μL。结果:盐酸黄酮哌酯和前体杂质3-甲基黄酮-8-羧酸的保留时间分别为7.0min和5.0min,分离度为6.0,最低检测浓度分别为1.20μg·mL~(-1)和0.55μg·mL~(-1)。盐酸黄酮哌酯在0.119～0.834mg·mL~(-1)范围内呈良好的线性关系,r=0.9999。平均回收率(n=5)分别为99.04％(RSD=0.66％),99.36％(RSD=0.87％),99.64％(RSD=0.62％)。结论:本方法简便、快速、专属,结果准确可靠,能有效控制产品质量。     

高效液相色谱法测定桂枝中3种有效成分的含量

目的:建立HPLC法测定桂枝中桂皮醛、栓皮酸和邻甲氧基栓皮醛的含量。方法:采用ODS色谱柱(150mm×4.6mm,5μm),以甲醇-水-冰乙酸(45:55:0.05)为流动相,流速为0.8 mL·min~(-1),紫外检测波长为280 nm。结果:栓皮醛、桂皮酸和邻甲氧基桂皮醛的线性范围(n=5)分别为0.125～2μg(r=0.999 8),0.003 2～2 μg(r=0.999 9),0.012 5～0.2μg(r=0.999 7),平均回收率(n=9)分别为99.3％,102.8％,98.9％。提取方法为甲醇冷浸12h。结论:本方法简便、准确,可为评价不同产地的桂枝质量提供依据。     

RP-HPLC法研究西红花酸在大鼠体内的药代动力学和组织分布特性

目的:建立大鼠血浆中西红花酸的RP-HPLC分析方法,并对其大鼠体内过程特性进行分析研究。方法:生物样品经甲醇沉淀蛋白质并提取药物,采用RP-HPLC法,色谱柱:Kromasil C18反相柱(150 mm×4.6mm,5μm);流动相为甲醇-水-冰醋酸(74:24:2);流速1.0 mL·min-1;柱温30℃;检测波长423nm。结果:方法回收率为(93.08±2.4)％-(106.5±7.1)％。测定血药浓度线性范围为0.4903-15.36μg·mL-1(r=0.9995),日内RSD<5％,日间RSD<8％,最低检测限13.4ng·mL-1(S/N=3)。SD大鼠一次灌胃给药西红花酸后血药浓度-时间曲线呈二室模型,半衰期t1/2(66.3±2.2)min。在主要组织脏器分布特点是c肝>C肺>C子宫、卵巢>C肾>C脂肪>C睾丸。结论:本法准确、灵敏度较高,可用于西红花酸体内过程的研究。西红花酸主要由肾脏原型排泄。     

高效液相色谱-蒸发光散射检测法测定中药及制剂中齐墩果酸和熊果酸的含量

目的:建立高效液相色谱-蒸发光散射检测法测定中药饮片及中成药制剂中齐墩果酸和熊果酸的含量。方法:采用Kromasil C18(4.6 mm×250 mm,5μm)色谱柱,柱温40℃;甲醇-水-冰醋酸(260:40:0.15)为流动相,流速0.6 mL·min-1;蒸发光散射检测器检测,漂移管温度85℃,气体流速2.60 L·min-1。结果:齐墩果酸在0.562-2.81μg范围内(r=0.9998),熊果酸在0.380-1.90μg范围内(r=0.9996)线性关系良好。药材(木瓜)中齐墩果酸和熊果酸的回收率分别为98.2％和99.8％,RSD(n=5)分别为3.2％和3.4％。中成药制剂(知柏地黄丸)中齐墩果酸和熊果酸的回收率分别为100.0％和100.1％,RSD(n=5)分别为2.1％和2.9％。结论:方法简便、准确,重现性好。     

HPLC法测定麻仁丸中大黄素和大黄酚的含量

目的:建立麻仁丸中大黄素和大黄酚含量测定的方法。方法:色谱柱为Thermo ODS C18柱(250 mm×4．6 mm,5μm),流动相为甲醇-0．1％磷酸溶液(85:15),流速为1．0 ml·min-1,检测波长为254 nm。结果:大黄素在0．01-0．29μg(r =0.999 5);大黄酚在0．03-0．52μg(r=0．999 4)范围内线性关系良好。大黄素及大黄酚的平均回收率分别为100.4％(RSD =0．9,n=5),100．1％(RSD=0．3％,n=5)。结论:方法准确可靠。     

RP-HPLC法测定桂枝中4种活性成分的含量

目的:建立了同时测定桂枝中桂皮醛、桂皮酸、桂皮醇及香豆素含量的方法,考察了4种活性成分的含量与产地及采收季节的关系。方法:采用反相高效液相色谱法,以Luna C18(4.6mm×250mm,5 μm)为色谱柱,乙腈-0.1％磷酸(28:72)为流动相,检测波长285nm,流速1mL·min-1,柱温30℃。结果:桂皮醛、桂皮酸、桂皮醇及香豆素的线性范围为0.208-3.33μg(r=0.9997),0.100-1.60μg(r=0.9999),0.204-3.26μg(r=0.9999),0.0674-1.08μg(r=0.9999);回收率分别为98.3％(RSD=1.0％),98.0％(RSD=1.4％),99.1％(RSD=1.2％),98.3％(RSD=1.8％)。结论:本法简便、准确、重现性好,适用于桂枝中4种成分的同时测定;生长环境和采收季节对桂枝中4种活性成分的含量有显著影响。     

人血浆中富马酸奎硫平浓度的测定

目的:建立测定人血浆中富马酸奎硫平(QTP)浓度的HPLC法。方法:以Diamonsil~(TM)C_(18)柱(250mm×4.6mm,5μm)为色谱柱,流动相:甲醇-超纯水(85:15,V/V),流速为0.8mL·min~(-1),检测波长254nm,以乙酸乙酯为提取剂。结果:QTP 80.00,16.67,1.67μg·mL~(-1)高、中、低3个浓度的平均回收率分别为98.20％,101.44%,98.21％。日内、日间RSD均<5%(n=5)。分析方法的最小检测限为0.08μg·mL~(-1)(r_(SN)=2)。线性范围为0.17～100.00μg·mL~(-1),线性回归方程为Y=8.78×10~(-2)X-3.37×10~(-2),r=0.9999(n=11)。结论:该方法灵敏、准确、简单、快速,可用于临床血药浓度监测和药动学研究。     

诺司咪唑含量测定方法的探讨

目的:建立了测定诺司咪唑含量的高效液相色谱法和胶束电动毛细管色谱法。方法:高效液相色谱法的实验条件:选用Dikma Diamonsil~(TM)(钻石)C_(18)色谱柱(4.6 mm×250 mm,5μm),乙腈-0.05mol·L~(-1)磷酸二氢钠水溶液(42:58,含0.2％三乙胺,pH 3.0),流速:1mL·min~(-1),检测波长:278 nm;进样量:20μL;柱温:30℃;胶束电动毛细管色谱法的实验条件:采用石英毛细管柱(未涂层)68.5 cm×75μm,有效柱长60 cm,优化选择40 mmol·L~(-1)磷酸二氢钠+25mmol·L~(-1)SDS(pH 6.0)为电泳介质,操作电压25 kV,柱温:30℃,检测波长:278 nm。结果:本法简便、灵敏、准确。两法线性范围分别为:0.8～30.0μg·mL~(-1)和27～269 μg·mL~(-1),回归方程分别为A=-602.7+5 334.0C(r=0.999 9)和A=0.007 9+0.174C(r=0.999 7),高效液相色谱法的平均回收率为99.8％(n=5)。结论:本文建立的方法可用于诺司咪唑的含量测定。     

中药材木通质量评价方法的研究

目的:建立TLC和HPLC法,同时对木通中齐墩果酸和常春藤皂苷元进行定性、定量分析。方法:薄层鉴别采用硅胶G薄层板,展开剂:正己烷-醋酸乙酯-冰醋酸(6:4:0.25),显色剂:10％硫酸乙醇溶液;含量测定采用Kromasil C18色谱柱(4.6mm×150 mm,5 μm),流动相:甲醇-水-冰醋酸-三乙胺(87:13:0.04:0.02),检测波长:210 nm,流速:0.8 mL·min-1,柱温:室温。结果:TLC色谱系统中,齐墩果酸与常春藤皂苷元分离效果良好;HPLC中齐墩果酸、常春藤皂苷元线性范围分别为2.03-50.8μg(r=0.9996)和2.08-52.1μg(r=0.9997),平均回收率(n=5)分别为99.1％(RSD=1.8％)和97.4％(RSD=2.0％)。结论:所建立的TLC和HPLC法重现性好,专属性强,为木通药材质量控制提供科学依据。     

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.     

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.