葛根芩连汤不同制法对葛根素含量的影响

目的 :考察用煎药机与传统方法制得葛根芩连汤汤剂中 ,葛根素含量的变化。方法 :采用HypersilC1 8(5 μm,ID 4.6× 2 5 0mm )分析柱 ,流动相为甲醇 水 (3 0∶70 ) ,柱温 2 5℃ ,流速 1ml·min- 1 ,检测波长 2 5 0nm。结果 :采用传统方法煎煮得到的葛根芩连汤中 ,葛根素的含量为 166.97mg;采用煎药机煎制得的葛根芩连汤中 ,葛根素的含量为 112 .70。结论 :采用传统方法煎煮制得的汤剂中 ,葛根素含量较高     

HPLC法同时测定葛根芩连片中葛根素和黄芩苷的含量

目的 建立同时测定葛根芩连片中的葛根素和黄芩苷含量的方法.方法 采用高效液相色谱法,使用Ultimate LP-C18色谱柱;流动相:甲醇-1％冰醋酸溶液;检测波长:0～20min,250nm,20～45min,278nm;流速:1.0mL?min-1;柱温:35℃.结果 葛根素在0.03869～0.7738μg(r=0.99999)范围内,黄芩苷在0.02052～0.4104μg(r=0.99999)范围内,线性关系良好.葛根素平均回收率为98.57％,RSD=1.6％;黄芩苷平均回收率为99.41％,RSD=1.9％.结论 本法专属性强、准确度高,可作为葛根芩连片质量控制的方法.     

UPLC快速测定葛根芩连汤肠外翻囊样品中6个黄酮类成分含量

目的:建立UPLC法同时测定葛根芩连汤肠外翻囊样品中6个黄酮类成分(葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷)的含量。方法:样品用甲醇提取,采用Agilent Proshell 120 EC-C18(4.6 mm×50 mm,2.7μm)色谱柱,以0.2%醋酸水溶液(A)-甲醇(B)为流动相进行梯度洗脱,流速1 mL.min-1,紫外检测波长270 nm,柱温30℃。结果:葛根素、大豆苷、甘草苷、野黄芩苷、黄芩苷、汉黄芩苷质量浓度分别在0.1870～93.50μg.mL-1(r=0.9999),0.02730～13.65μg.mL-1(r=0.9999),0.02550～12.75μg.mL-1(r=0.9999),0.05580～27.90μg.mL-1(r=0.9998),0.2124～106.2μg.mL-1(r=0.9999),0.09140～45.70μg.mL-1(r=0.9999)范围内有良好的线性关系;日间、日内精密度变异均小于2%;平均回收率(n=9)分别为101.4%,97.88%,99.15%,99.72%,98.50%,101.4%;稳定性良好。结论:所建立的分析方法操作简便、快速、灵敏,结果准确,重复性好,所需样品少,并已成功应用于葛根芩连汤大鼠肠吸收动力学的研究。     

HPLC测定葛根芩连汤不同煎液中葛根素的含量

目的 :考察用合煎、分煎和沸水冲泡单味中药精制颗粒等方法制备的葛根芩连汤汤剂中葛根素含量的变化。方法 :采用HypersilC18柱 (5 μ ,id 4. 6mm× 2 5 0mm) ,流动相甲醇 水 (30∶70 ) ,流速 1ml·min 1,柱温2 5℃ ,检测波长 2 5 0nm ,外标法定量。结果 :葛根芩连汤分煎液中葛根素的平均含量为 2 8. 74mg ,合煎液中平均含量为 2 3. 0 1mg,单味中药精制颗粒冲泡液中平均含量为 13. 37mg。结论 :用不同方法制备的汤剂中葛根素的含量有差异     

心安宁胶囊质量标准研究

目的　建立心安宁胶囊质量标准。方法　建立了葛根的显微鉴别方法和葛根、何首乌的薄层鉴别方法 ,并采用HPLC法测定其中葛根素的含量 ;色谱条件为 :C18柱 ;流动相 :甲醇 水 (2 2∶78) ;流量 :1 0ml/min ;检测波长 :2 5 0nm ;柱温 :室温。结果　本方法简便准确 ,重复性好。葛根素在 0 0 0 4～ 0 16 3μg范围内线性关系良好 (r =0 9998) ,平均回收率为 99 4 % (n =6 ) ,RSD为 2 4 %。结论　新建质量标准可有效地控制心安宁胶囊质量。     

葛根芩连片的质量控制研究

目的以有效部位为指标,研究葛根芩连片质量控制新方法。方法采用紫外分光光度法(UV)测定总黄酮含量;采用高效液相色谱法(HPLC)测定葛根素含量,菲罗门C18色谱柱(4.6mm×250mm,5μm),流动相为甲醇-水(28:72),流速为1.0ml/min,柱温30°C,检测波长为250nm。结果总黄酮含量为12.26%～12.73%(RSD为1.21%);葛根素含量为2.24%～2.43%(RSD为2.93%)。结论葛根总黄酮含量间变异比葛根素小,更适合作为质量控制指标。     

RP-HPLC法测定葛根芩连片中葛根素的含量

目的:建立以RP-HPLC法测定葛根芩连片中葛根素含量的方法。方法:色谱柱为Zorbax SB-C18(250mm×4.6mm,5μm),流动相为甲醇-水-冰醋酸(23∶77∶1),检测波长为250nm。结果:葛根素在0.003916~0.3916mg·mL-1浓度范围内与峰面积积分值呈良好的线性关系(r=0.9999);平均回收率为100.40%,RSD=0.42%(n=9)。结论:本方法简便、快速、准确,可用于该制剂的质量控制。     

高效液相色谱法测定不同产地葛根中葛根素的含量

目的建立测定葛根中葛根素含量的RP-HPLC方法,研究不同地区葛根中葛根素的含量差异。方法采用反相高效液相色谱法,Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为甲醇-水(25∶75),流速为1.0 mL.min-1,检测波长为250 nm,柱温为30℃。结果葛根素在0.071~1.42μg与峰面积呈良好的线性关系(r=0.999 9),平均回收率为98.4%,RSD为1.5%(n=6)。结论本方法简便、准确,重现性好,适用于葛根中葛根素含量的测定。不同产地葛根中葛根素含量差异较大。     

玉泉胶囊质量标准研究

目的:建立玉泉胶囊的质量标准。方法:采用薄层色谱(TLC)法定性鉴别方中葛根、人参、甘草、黄芪、五味子;采用高效液相色谱(HPLC)法测定葛根素含量。结果:TLC法分别检测出玉泉胶囊中葛根、人参、甘草、黄芪、五味子;葛根素进样量在0.1275～1.02μg范围内与峰面积积分值呈良好的线性关系(r=0.9999),平均回收率为100.29%,RSD=1.68%。结论:所建标准能有效控制玉泉胶囊的质量。     

葛根芩连汤氯仿部位含量测定及对羧酸酯酶的抑制作用研究

目的:建立葛根芩连汤氯仿部位的多成分含量测定方法,并考察其抑制羧酸酯酶(CES2)的作用。方法:采用溶剂萃取法提取葛根芩连汤氯仿部位,以葛根素、小檗碱、黄芩苷、甘草苷4种指标性成分,采用超高效液相色谱(UPLC)对其进行含量测定;采用荧光素二乙酸酯(FD)为CES2的特异性荧光探针底物,在人肝微粒体(HLM)孵育体系中考察葛根芩连汤氯仿部位对CES2的抑制作用。结果:葛根芩连汤氯仿部位中指标性成分含量测定结果为:葛根素347 mg.g~(-1)、小檗碱1.65 mg.g~(-1)、黄芩苷14.9 mg.g~(-1)、甘草苷1.04 mg.g~(-1),该部位抑制CES2活性的IC_(50)为0.5 mg.mL~(-1)。结论:葛根芩连氯仿部位及其单体均对CES2具有较强的抑制作用,揭示葛根芩连汤有效部位可通过抑制羧酸酯酶以缓解伊立替康所致迟发型腹泻,为中药缓解其毒副作用的机制提供科学依据。     

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.

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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.     

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.