黄连提取工艺的研究

采用硅胶G薄层色谱法对黄连进行鉴定,证明黄连的提取物中主要成分为小檗碱。运用正交试验设计法筛选出黄连中小檗碱提取的最佳条件,采用高效液相色谱法,以提取液中的小檗碱的含量为指标优选出黄连中小檗碱的提取工艺。本实验表明,回流提取或连续回流提取,水(用黄连粗粉,回流2次,每次1.5小时,加水量为12倍)和乙醇(体积浓度分别为50%、60%、70%)提取黄连中小檗碱的效果基本相同。因此,用水代替乙醇作溶剂,既可保证提取的质量,又能降低成本。本研究结果为黄连中小檗碱的工业化生产提供了理论参考。     

毛细管电泳法测量清热化毒丸中盐酸小檗碱的含量

目的建立毛细管电泳法测量清热化毒丸中盐酸小檗碱含量的方法。方法采用甲醇溶液提取清热化毒丸样品,通过高效毛细管电泳法,测量盐酸小檗碱的含量。熔融石英毛细管柱（47cm×75m,有效长度40cm）;缓冲液体系为：60mmol／L磷酸缓冲溶液．甲醇（65：35）,pH：3．2;分离电压30KV;毛细管柱温25℃;检测波长200nm;进样时间5s。结果盐酸小檗碱在进样浓度为0．05mg／ml～0．10mg／ml内线性关系良好（r=0．9993）,平均加样加收率为103．27％,RSD为0．87％。结论该方法简便、快捷、灵敏度高、重现性好,可用于清热化毒丸中小檗碱含量的测定。     

正交试验优选黄连中小檗碱提取工艺的研究

目的 :筛选黄连中小檗碱的提取工艺。方法 :采用正交试验设计系统考察乙醇浓度、乙醇用量、硫酸加入量、提取时间、提取次数5因素对盐酸小檗碱得率的影响。结果 :黄连中小檗碱适宜的提取工艺 :溶剂为6倍量80 %乙醇 ,乙醇中硫酸加入量为0 25 % ,提取时间为每次1 5h ,提取次数为3次。结论 :以该工艺制备的盐酸小檗碱精制品含量在90 %以上 ,适合工业化生产。     

正交试验优化提取黄连中小檗碱型生物碱

目的:研究黄连中小檗碱型生物碱的最佳提取工艺。方法:采用正交试验设计,以小檗碱、巴马汀、黄连碱、表小檗碱和药根碱的总含量为考察指标,考察乙醇浓度、提取时间、提取次数、乙醇用量4个因素对提取结果的影响。结果:最佳提取工艺为8倍量的50%乙醇,回流提取4次(每次2h)。结论:本工艺重现性好,测定方法可行,为大批量黄连提取生物碱提供了理论依据。     

结肠安肠溶片中小檗碱的提取工艺研究

目的利用高效液相色谱法优化结肠安肠溶片中小檗碱的提取工艺。方法以干膏率和干膏中小檗碱的含量为指标,通过正交试验筛选最佳提取工艺。结果最佳工艺条件为乙醇浓度为85%,每次提取2次,溶剂量6倍,提取时间3 h。结论醇提法可用于黄柏中有效成分的提取。     

黄连中生物碱提取工艺的正交试验研究

建立了HPLC法测定黄连及其提取物中的盐酸小檗碱（1）、提取工艺各影响因素。结果显示，药材用60％乙醇回流提取3次为22．06％、5．28％0和6．90％盐酸巴马亭（2）和盐酸黄连碱（3）。应用正交试验法考察每次1h，提取物得率25．47％；其中1、2、3的含量分别     

不同提取方法对黄连中小檗碱含量的影响

目的:探讨不同提取方法对黄连中小檗碱含量的影响。方法:用药典法、回流法、超声法、回流超声法4种方法提取黄连,以高效液相色谱法分别测定提取物中小檗碱的含量。结果:《中国药典》提供的方法与其它提取方法所得小檗碱含量结果有显著性差异(P<0·01)。结论:回流超声法与其它方法比较,时间短、含量高,较适宜提取黄连中的小檗碱。     

两种方法制备黄连解毒汤中小檗碱含量的比较

目的：探讨高温煎煮对黄连解毒汤中小檗碱含量的影响。方法：建立黄连解毒汤汤剂中小檗碱的高效液相色谱法(HPLC)含量测定法，比较GNG中药抽出机煎煮法与常压直火煎煮法制备的黄连解毒汤汤剂中小檗碱含量的差异。结果：GNG中药抽出机制备的黄连解毒汤汤剂中小檗碱含量明显小于常压直火煎煮法，两者有显著差异。结论：含小檗碱成分的中药不适合采用GNG中药抽出机煎煮制备。     

高效液相色谱法测定香连片中小檗碱的含量

目的 建立高效液相色谱法测定香连片中小檗碱含量的方法。方法 固定相为Hypersil C18柱；流动相为乙腈．水(45：55)，每1000ml加十二烷基磺酸钠1．0g；检测波长为254nm。结果 该方法的线性范围为8～24μg(r=0．9998，H=6)，平均回收率为99．9％，RSD=0．3％。结论 本方法简便，准确，灵敏度高，重复性好，可用于该片荆的成分含量测定。     

非共价涂层毛细管电泳法测定黄连及其炮制品中3种生物碱的含量

目的建立同时测定黄连药材中小檗碱、巴马汀和药根碱含量的毛细管电泳方法。方法采用聚凝胺-聚丙乙烯磺酸钠涂层毛细管柱(33 cm×50μm I.D.,有效长度为24.7 cm),运行缓冲液为500 mmol.L-1Tris-甲醇(体积比为70∶30)缓冲溶液(8 mol.L-1,磷酸调节pH 7.5),分离电压为22 kV,检测波长为230 nm,温度为20℃,压力自动进样(5 000 Pa×6 s),阴极检测。结果在所选电泳条件下,黄连中的生物碱组分在4.5 m in内得到良好分离,盐酸小檗碱、盐酸巴马汀和盐酸药根碱的线性范围分别为199.8~1 998、48.45~484.5和47.3~473 mg.L-1,线性关系良好,加样回收率分别为102.8%、100.2%和97.7%。结论该方法可应用于黄连药材中生物碱的分析测定。     

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.

---

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.