高效液相色谱-蒸发光散射检测法测定银杏叶片中4种萜类内酯含量

目的 优化银杏叶片银杏萜类内酯A、B、C及白果内酯含量测定方法.方法 采用高效液相色谱-蒸发光散射检测法(HPLC-ELSD),色谱柱:Purispher star-C18(4.6 mm×250 mm,5 μm),以A:B(31:69)[A:四氢呋喃-乙腈-甲醇(5:22:12),B:1%醋酸溶液]为流动相,柱温为室温,流速1 mL·min-1,载气为氮气,压力350 kPa,漂移管温度40 ℃,增阈值6.0.结果 银杏内酯C进样量在0.46～13.80 μg范围内线性关系良好,r=0.999 0;平均回收率为99.14%(n＝5),RSD＝1.39%;白果内酯进样量在1.175～35.250 μg范围内线性关系良好,r=0.999 7,平均回收率98.75%(n＝5),RSD＝2.06%.银杏内酯A进样量在0.875～26.250 μg范围内线性关系良好,r=0.999 0,平均回收率98.71%(n＝5),RSD＝3.11%;银杏内酯B进样量在0.475～14.250 μg范围内线性关系良好,r=0.999 1,平均回收率97.80%(n＝5),RSD＝3.22%.结论 优化后的方法简便,快速,准确,稳定性和重复性较好,可用于银杏叶片萜类内酯的含量测定.     

高效液相色谱法-蒸发激光散射检测器测定银杏酮酯舌下片中萜类内酯的含量

目的 :采用高效液相色谱法 -蒸发激光散射检测器测定银杏酮酯舌下片中总内酯的含量 ,以控制该制剂质量。方法 :应用高效液相色谱 -蒸发激光散射检测法分析 4种萜类内酯的含量。色谱条件为 :Diamonsil TM (钻石 ) C1 8色谱柱(2 5 0 mm× 4 .6 mm,5μm) ;流动相 :甲醇 -水 (36 :6 4 ) ;柱温 :30 o C;流速为 0 .6 ml/ min;检测器条件 :漂移管温度为 4 0 o C,载气压力为 2 .0 bar,灵敏度为 6。结果 :4种萜类内酯之间分离度均达 2 .5以上 ;理论板数以白果内酯峰计算为 830 8,6次平均加样回收率分别为 :白果内酯 10 2 .3% (RSD为 1.4 9% ) ,银杏内酯 C 10 2 .6 % (RSD为 1.31% ) ,银杏内酯 A 10 1.2 % (RSD为 2 .39% ) ,银杏内酯 B98.0 % (RSD为 2 .75 % ) ;白果内酯、银杏内酯 C、银杏内酯 A、银杏内酯 B,分别在 3.4～ 2 0 .4 μg、2 .2 2～ 13.32μg、2 .4 6～ 14 .76μg、2 .0 6～ 12 .36μg范围内 ,进样量对数值与峰面积对数值呈良好的线性关系。结论 :本法测定银杏酮酯舌下片中萜类内酯的含量 ,结果准确 ,重复性好     

HPLC-RI法测定银杏叶提取物中4种内酯的含量

目的:建立高效液相色谱法测定银杏叶提取物中4种内酯(银杏内酯 A、银杏内酯 B、银杏内酯 C、白果内酯)的含量。方法:采用 Symmetryshield 型 C_(18)色谱柱(3.9 mm×150 mm,5 μm),以甲醇-水(30:70)为流动相,流速为1.0 mL·min~~(-1),用外标法测定,示差折光检测器检测。结果:4种内酯浓度在0.02～0.24 mg·mL~(-1)范围内,与色谱峰面积呈良好线性关系,相关系数在0.9984～0.9998之间,检测限在0.025～0.035 μg之间,回收率在99.0%～100.2%之间。结论:本方法简单、准确、可靠、灵敏,适用于实验室和企业常规质量控制。     

高效液相色谱法测定银杏内酯B滴丸的含量

目的建立银杏内酯B滴丸中银杏内酯B的含量测定方法。方法采用HPLC,选择Ec lipseXDB-C18(150mm×4.6mm,5μm)色谱柱,以甲醇-0.1%磷酸溶液(33.3∶66.7)为流动相;流速:1.0mL.m in-1;检测波长:225nm。结果银杏内酯B在0.2650~4.240mg.mL-1范围内线性关系良好(r=1.000);平均加样回收率为99.6%(RSD=1.3%)。结论此方法简便、快速、准确,可作为银杏内酯B滴丸的含量测定方法。     

银杏酮酯分散片萜类内酯的溶出度试验

目的建立银杏酮酯分散片萜类内酯的溶出度测定方法.方法以萜类内酯为指标,采用高效液相色谱法-蒸发光检测法进行检测,采用ZORBAX RX-C18色谱柱(4.6mm×150 mm,5nμm);以水-甲醇-四氢呋喃(752010)为流动相;流速1.0mL·min-1.结果白果内酯、银杏内酯A、银杏内酯B、银杏内酯C的线性范围分别为1.519～7.595、1.087～6.522、1.01～3.03、1.056～3.168μg,平均回收率分别为99.55%(RSD为1.87%);99.79%(RSD为1.94%);99.78%(RSD为2.38%);98.21%(RSD为2.54%).结论该方法重复性较好,可作为银杏酮酯分散片的溶出度测定方法.     

高效液相色谱法测定银杏叶缓释胶囊中银杏内酯的含量

目的:建立高效液相色谱法,测定银杏叶缓释胶囊中银杏内酯的含量.方法:色谱柱:C18(Sperisorb 4.6 mm×250 mm,5 μm),流动相:甲醇-水(30∶70),流速:1.0 mL*min-1,柱温:25℃,示差折光检测器.结果:白果内酯、银杏内酯A、银杏内酯B、银杏内酯C进样量与色谱峰面积在10～80 mg*L-1范围内呈良好的线性关系, R分别为0.9996,0.9995,0.9999,0.9997,回收率分别为100.0%,99.1%,99.2%,99.9%,精密度测定RSD小于6.6%.结论:该方法灵敏、准确、经济,可用于测定银杏叶缓释胶囊中银杏内酯的含量.     

HPLC-CAD法测定银杏叶片中萜类内酯的含量

目的建立银杏叶片中萜类内酯的含量测定方法。方法采用HPLC-CAD法测定白果内酯、银杏内酯A、银杏内酯B和银杏内酯C的含量。使用Agilent TC C_(18)色谱柱(250 mm×4.6 mm,5μm),以正丙醇-四氢呋喃-水(1∶15∶84)为流动相,流速1.0 mL·min~(-1),柱温40℃,采用电喷雾检测器进行检测。结果各萜类内酯在线性范围内与峰面积线性关系良好,相关系数为0.9997～1.000,平均回收率(n=6)在97.3%～99.1%,RSD在1.6%～2.5%。结论该方法灵敏度高,精密度、重复性、准确度均良好,可作为银杏叶片中萜类内酯的含量测定方法。     

银杏叶片和银杏酮酯片的质量比较和用药成本分析

目的 比较市售3个厂家银杏叶片和银杏酮酯片、2个厂家银杏叶提取物和银杏酮酯中总黄酮、总黄酮醇苷和萜类内酯的含量,为评价2种制剂的质量提供依据;并分析2种银杏叶制剂的药物经济学。方法 通过交叉试验,采用UV检测2种制剂和原料药中总黄酮的含量,采用HPLC检测2种制剂和原料药中总黄酮醇苷和萜类内酯的含量。结果 2种制剂和原料药分别在总黄酮、总黄酮醇苷和萜类内酯的含量上均不存在显著性差异,但就患者日用金额考虑,银杏叶片更适合患者使用。结论 银杏叶片较银杏酮酯片更适合用于临床上慢性疾病的治疗。     

不同类型银杏叶中银杏内酯的含量测定

目的 :建立HPLC法测定各种类型银杏叶中银杏内酯的含量。方法 :色谱柱为SUNIERKromasilC1 8(5 μm ,4 6mm×2 5 0mm) ,以 33%甲醇为流动相 ,流速 :1mL·min- 1 ,示差检测器 (RI) ,以银杏内酯A、B、C及白果内酯 4种对照品为对照。结果 :测定了各种类型的银杏叶内酯的含量。结论 :找出了银杏叶的最佳采集时间。     

蒸发光散射检测法测定银杏叶片中4种萜类内酯含量

目的优化银杏叶片银杏萜类内酯A、B、C及白果内酯含量测定方法。方法采用高效液相色谱 蒸发光散射检测法（HPLC ELSD）,色谱柱：Purispher star C18（4.6 mm×250 mm,5 μm）,以A：B（31：69）[A：四氢呋喃 乙腈 甲醇（5：22：12）,B：1%醋酸溶液]为流动相,柱温为室温,流速1 mL•min 1,载气为氮气,压力350 kPa,漂移管温度40 ℃,增阈值6.0。结果银杏内酯C进样量在0.46～13.80 μg范围内线性关系良好,r=0.999 0;平均回收率为99.14%（n＝5）,RSD＝1.39%;白果内酯进样量在1.175～35.250 μg范围内线性关系良好,r=0.999 7,平均回收率98.75%（n＝5）,RSD＝2.06%。银杏内酯A进样量在0.875～26.250 μg范围内线性关系良好,r=0.999 0,平均回收率98.71%（n＝5）,RSD＝3.11%;银杏内酯B进样量在0.475～14.250 μg范围内线性关系良好,r=0.999 1,平均回收率97.80%（n＝5）,RSD＝3.22%。结论优化后的方法简便,快速,准确,稳定性和重复性较好,可用于银杏叶片萜类内酯的含量测定。     

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.