HPLC法测定绞股蓝中总槲皮素、山柰素和异鼠李素的含量

目的：建立测定绞股蓝中槲皮素、山柰素和异鼠李素含量的HPLC方法。方法：色谱柱为Hypersil ODS2（250mm×4．6mm,5μm）;流动相为甲醇-0．4％磷酸水溶液（50：50,V／V）;流速0．78ml·min^-1;检测波长为360nm;柱温为25℃。结果：槲皮素、山柰素和异鼠李素均可达基线分离,其线性范围分别为20．02～100．10μg·ml^-1（r=0．9999）,12．16～60．80μg·ml^-1（r=0．9999）,1．45～7．26μg·ml^-1（r=0．9998）。平均加样回收率分别为97．32％（RSD=1．09％）,97．75％（RSD=1．06％）,97．29％（RSD=1．29％）。结论：该方法简单、快速、准确,可用于绞股蓝药材的质量控制。     

西河柳中槲皮素与异鼠李素在植物体内分布及随季节变化

目的:建立西河柳中槲皮素与异鼠李素的HPLC测定方法;研究槲皮素与异鼠李素在西河柳不同部位的分布及随季节变化的规律。方法:以NUCLEODUR100-5C_(18)柱(150 mm×4.6 mm,5μm),甲醇-0.4%磷酸溶液(50:50)为流动相,流速为0.8ml·min~(-1),柱温为30℃,检测波长为360 nm。结果:槲皮素与异鼠李素线性范围分别为:0.17～1.01μg(r=0.999 7)和0.19～0.95μg(r=0.999 6),回收率分别为:102.3%,100.7%RSD4.0,3.1%;西河柳中槲皮素与异鼠李素含量随季节的变化而变化,且不同部位有不同的含量。结论:该方法可用于控制西河柳药材质量;西河柳中槲皮素与异鼠李素含量按春、夏、秋次序依次降低,在嫩技叶中含量最高。     

高效液相色谱分离柱后化学发光法同时测定菟丝子中黄酮类成分

目的:建立高效液相色谱分离柱后化学发光法测定菟丝子中芦丁、山柰素、槲皮素和异鼠李素含量。方法:基于在氢氧化钠碱性介质中铁氰化钾可以直接氧化芦丁、山柰素、槲皮素和异鼠李素产生化学发光。色谱柱:Hypersil ODS(5μm,4.6 mm×150 mm),流动相:乙醇-乙腈-水-磷酸(21:22:55:2),流速:1mL·min~(-1),柱温:25℃。结果:芦丁、山柰素、槲皮素和异鼠李素浓度分别在0.2～5μg·mL~(-1)(r=0.9990),0.1～15μg·mL~(-1)(r=0.9991),0.1～100μg·mL~(-1)(r=0.9994),0.3～200μg·mL~(-1)(r=0.9998)范围内,与峰面积有良好的线性关系;检测限(S/N=3)分别为0.02,0.08,0.08,0.03μg·mL~(-1)。结论:水法简便、快速、有效,灵敏度高,首次用于菟丝子中黄酮类成分的测定,取得了很好的结果。     

HPLC法同时测定南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量

目的 建立南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量测定方法。方法 采用Diamond—C18色谱柱;甲醇-0．4％磷酸溶液（50：50）为流动相;流速1．0mL·min;检测波长360nm。结果 线性范围：槲皮素0．1184～0．8288μg,r=0．9995,山奈酚0．0258～0．1806／μg,r=0．9994,异鼠李素0．0444～0．3108μg,r=0．9994;平均回收率：槲皮素99．51％,RSD=2．10％（n=6）,山奈酚101．03％,RSD=2．08%（n=6）,异鼠李素99．80％,RSD=2．09％（n=6）。结论 该法操作简便、准确,重复性好,可用于南葶苈子饮片中槲皮素、山奈酚及异鼠李素的含量测定。     

HPLC法测定垂盆草配方颗粒中黄酮类成分的含量

目的:建立高效液相色谱法测定垂盆草配方颗粒中槲皮素、山奈素及异鼠李素含量的方法.方法:流动相为甲醇:0.4％磷酸溶液(45∶ 55),色谱柱为Diamonsil C18(200 mm× 4.6 mm,5μm),检测波长为360 nm,柱温为30℃,流速为1.0ml·min-1.结果:三种黄酮类成分分离度好,不受杂质干扰,线性范围:槲皮素在2.982 ～29.820 μg· ml-1,r =0.9997,山奈素在0.966～9.660μg·ml-1,r =0.9998,异鼠李素在0.982～9.820 μg·ml-1,r =0.9994;平均回收率:槲皮素99.26％,RSD=0.66％(n=6),山奈素99.21％,RSD=1.24％ (n =6),异鼠李素99.44％,RSD =0.40％(n=6).结论:该测定方法操作简便,快捷,可靠性高,重复性好,可作为垂盆草配方颗粒的质量控制方法之一.     

高效液相色谱法同时测定田基黄药材中槲皮素、芦丁和异鼠李素的含量

目的建立高效液相色谱法同时测定田基黄药材中槲皮素、芦丁和异鼠李素的含量。方法采用HPLC法,色谱柱为Agilct Hypcrsil C18（4．6mm×250mm,5μm）,甲醇-0．2％磷酸（54：46）为流动相,流速为1mL·min^-1,检测波长264mm,柱温为室温。结果槲皮素、芦丁和异鼠李素分别在0．265-4．240、0．201～3．22、0.0755～1．208μg线性关系良好,平均回收率分别为99．7％0、99．7％、100．80,RSD分别为0．76%、1.28％、2．69％。结论所建立的方法简单易行,准确可靠,可用于田基黄药材中槲皮素、芦丁和异鼠李素的含量测定。     

三波长HPLC同时测定沙棘颗粒中的槲皮素、山萘酚与异鼠李素

目的 用HPLC法在3个波长段同时测定沙棘颗粒中槲皮素、山萘酚与异鼠李素的含量.方法 经室温两步提取处理沙棘颗粒中的黄酮类化合物,测定其中的槲皮素、山萘酚与异鼠李素.在反相液相系统中,采用Symmetry C18柱(250 mm×4.6mm,5.0 μm),流动相为乙腈-0.4％磷酸溶液(40∶60),室温,流速1.0 mL· min-1.槲皮素、山萘酚、异鼠李素的检测波长分别为266、360、370 nm.结果 槲皮素、山萘酚与异鼠李素均在0.2～1.6 μg与峰面积线性关系良好,平均回收率分别为99.7％、98.3％、101.0％,r分别为0.9993、0.9996、0.9991.结论 所用方法灵敏度高、专属性强,适用于测定复方沙棘颗粒中的槲皮素、山萘酚与异鼠李素的含量.     

超高效液相色谱法测定垂盆草颗粒中3种黄酮成分的含量

目的 建立超高效液相色谱法(UPLC)同时测定垂盆草颗粒中槲皮素、山柰素与异鼠李素的含量.方法 色谱柱Thermo Scientific AcclaimTM RSLC 120 C18(2.1 mm×100 mm,2.2 μm),流动相为甲醇-0.4％磷酸(50∶50),柱温为30℃,流速为0.3 mL·min-1,检测波长为360 nm,进样量1μL.结果 槲皮素、山柰素与异鼠李素的线性范围分别为:50～1 000 μg,20～400 μg,20 ～400 μg,且与峰面积呈良好线性关系(r=0.999 9,0.999 9,0.999 6),精密度、稳定性、重复性试验的RSD均＜2％,平均加样回收率(n=9)分别为99.35％,99.38％,98.91％,RSD分别为1.04％、0.98％、1.01％.结论 该法高效、准确、灵敏,可作为测定垂盆草颗粒中槲皮素、山柰素与异鼠李素含量的有效方法.     

银杏酮酯-羟丙基-β-环糊精包合物的质量标准研究

目的建立银杏酮酯-羟丙基-β-环糊精包合物的质量标准。方法采用高效液相色谱法测定制剂中的总黄酮苷含量,色谱柱:Diamonsil C18柱(250 mm×4.6 mm,5μm),流动相:甲醇-0.4%磷酸(45∶55),检测波长368nm,流速:1 mL.min-1;采用红外光谱分析法鉴别包合物。结果槲皮素在2~132μg.mL-1与峰面积具有良好的线性关系,r=0.999 5;山奈素在0.9~60μg.mL-1与峰面积具有良好的线性关系,r=0.999 8;异鼠李素在0.1~9μg.mL-1与峰面积具有良好线性,r=0.999 8。槲皮素、山奈素、异鼠李素的平均回收率分别为101.6%、100.4%、99.3%,RSD分别为1.3%,1.4%,0.63%。红外光谱法能有效鉴别包合物。结论本法可作为该制剂的质量控制参考标准。     

HPLC法测定五维赖氨酸颗粒中四种维生素的含量

目的:建立离子对RP-HPLC法测定五维赖氨酸颗粒中烟酰胺、维生素B6、维生素B2、维生素B1的含量。方法:用Zorbax Eclipe XDB-C18柱(250 mm×4．6 mm,5μm),流动相为0．005 mol·L-1庚烷磺酸钠溶液(含0．5％冰醋酸和0．05％三乙胺)-甲醇(72:28)为流动相,流速为1．0 ml·min-1,检测波长为280 nm。结果:烟酰胺、维生素B6、维生素B2及维生素B1的线性范围分别为0．32-15．77μg(r=0．999 7),0．05～2．61μg(r=0．999 9)、0．05-2．65μg(r=0．999 9)及0．45-22．37μg(r= 0．999 9),平均回收率分别为98．3％(RSD 1．3％,n=9)、101．1％(RSD 1．9％,n=9)、99．3％(RSD 1．0％,n=9)、98．9％(RSD 1．1％,n=9)。结论:本法简便,快速,可用于五维赖氨酸颗粒产品的质量控制。     

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.     

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.