HPLC法测定紫花前胡中紫花前胡苷的含量

目的:建立HPLC法测定中药紫花前胡中紫花前胡苷含量的方法,为其质量控制提供依据.方法:采用反相高效液相色谱法,Agilent Zorbax SB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为甲醇-水(40:60),流速为1mL·min-1,检测波长334 nm,柱温40℃.结果:紫花前胡苷在此色谱条件下获得良好分离,线性范围为25.0～300.0 ng(r2=0.9999),平均回收率为98.6%,RSD1.8%.结论:首次建立了两相系统HPLC法测定紫花前胡药材中紫花前胡苷含量的方法,该方法准确,可靠,对紫花前胡药材及其制剂的质量控制有较重要意义,已被2010版<中国药典>采纳.     

HPLC法测定白花前胡中白花前胡丙素和紫花前胡中紫花前胡苷的含量

目的 :建立高效液相色谱法测定白花前胡根中白花前胡丙素 (dl-praeruptorinA ,Pd -Ia)和紫花前胡根中紫花前胡苷 (nodakenin ,NDK)含量的方法。方法 :色谱柱Shim -PackCLC -ODS ,Pd -Ia的流动相为甲醇 -水 (79∶2 1)和NDK的流动相为乙腈 -甲醇 -水 (2 0∶5∶75 ) ,流速为 1 0mL·min- 1 ,紫外检测波长分别为 32 3和 334nm。结果 :Pd -Ia及NDK的线性范围分别为 0 0 8～ 0 8μg和 0 0 1～ 0 10 μg ,相关系数均为 0 9999,平均回收率分别为 98 5 3%～ 99 2 7%和 97 6 0 %～99 87% ,RSD小于 2 %。结论 :方法准确 ,简便 ,快速 ,灵敏度高 ,对常用中药前胡的质量控制有较重要意义。     

HPLC法测定白有胡中白花前胡丙素和紫花前胡中紫花前胡苷的含量

目的：建立高效液相色谱法测定白花前胡根中花前胡丙素（dl-praeruptorin A,Pd-Ia)和紫花前胡根中紫花前胡苷（modakenin,NDK)含量的方法。方法：色谱法Shim-Pack CLC-ODS,Pd-Ia的流动相为甲醇－水（79：21）和NDK的流动相为乙腈－甲醇－水（20：5：75）,流速为1．0mL.min^-1,紫外检测波长分别为323和334nm。结果：Pd-Ia及NDK的线性范围分别为0．08－0．8μg和0．01－0．10μg,相关系数均为0．9999,平均回收率分别为98．53％－99．27％和97．60％、99．87％,RSD小于2％。结论：方法准确,简便,快速,灵敏度高,对常用中药前胡的质量控制有较重要意义。     

紫花前胡化学成分的研究

目的研究中药紫花前胡［Peucedanum decursivum(Miq.)Maxim］根的化学成分。方法用硅胶柱色谱和制备型高效液相色谱进行分离纯化,用光谱分析和化学方法鉴定其结构。结果从紫花前胡根中分得7个线型吡喃香豆素类化合物,其结构分别鉴定为3′(S)-hydroxy-4′(R)-angeloyloxy-3′,4′-dihydroxanthyletin(1);3′(S)-acetoxy-4′(R)-hydroxy-3′,4′-dihydroxanthyletin(2);3′(S)-acetoxy-4′(R)-angeloyloxy-3′,4′-dihydroxanthyletin(3);Pd-C-IV(4);Pd-C-II(5);(+)-3′S-Decursinol(6);(+)-trans-Decursidinol(7)。结论化合物1和2为新化合物,分别命名为紫花前胡素D和紫花前胡素F;6和7为首次从该植物中分得。     

HPLC法筛查百花定喘制剂中前胡投料

目的： 通过测定白花前胡甲素、白花前胡乙素和紫花前胡苷的含量,建立筛查百花定喘制剂中投料为前胡还是紫花前胡的HPLC方法。方法：采用Symmentry-C18（4.6 mm×250 mm,5 μm）色谱柱,流动相：甲醇（A）-0.1%磷酸溶液（B）,梯度洗脱,检测波长为321 nm,流速为1.0 mL·min^-1。结果：5批药材的标示名称与试验判断结果不一致;47批百花定喘制剂的筛查结果表明,未见紫花前胡投料,前胡存在投料不足的情况。结论：该方法专属性良好,可作为定性筛查白花定喘制剂中投料前胡的方法。     

紫花前胡苷抗宫颈癌细胞作用研究

目的 探究紫花前胡苷抗宫颈癌HeLa细胞的作用效果及相关信号通路.方法 HeLa细胞在0、0.1、1和10 mmol/L浓度的紫花前胡苷作用24 h后,通过CCK-8法检测细胞增殖活性,流式细胞术检测细胞凋亡水平;通过蛋白免疫印迹法检测细胞内凋亡相关cleaved-caspase 3(cle-caspase 3)、Cytochrome C(Cyto C)、Bax、Bcl2蛋白水平变化,以及自噬相关p-mTOR、mTOR、P62、Beclin1、LC3蛋白水平变化;结合Ad-GFP-LC3B腺病毒转染检测细胞内LC3B聚集状态.结果 对照组(0 mmol/L)、0.1 mmol/L组、1 mmol/L组、10 mmol/L组的HeLa细胞凋亡率依次升高,分别为(14.93±2.95)％、(15.43±2.13)％、(21.63±3.19)％、(31.42±5.01)％,且在10 mmol/L浓度处理下升高最为明显(P<0.05).10 mmol/L的紫花前胡苷作用24 h后,HeLa细胞内的促凋亡cle-caspase 3、Cyto C、Bax蛋白水平升高,抗凋亡蛋白Bcl2表达下降,自噬相关蛋白p-mTOR/mTOR水平下降,Beclin1以及LC3-Ⅱ累积增多,自噬流相关蛋白P62表达下降,LC3B在紫花前胡苷处理组中呈颗粒性聚集,均差异有统计学意义(P<0.05).结论 紫花前胡苷具有较好的抗宫颈癌细胞作用,其机制可能与自噬相关细胞凋亡有关.     

紫花杜鹃药材质量标准研究

目的 研究制定紫花杜鹃药材的质量标准.方法 采用薄层色谱法对紫花杜鹃药材中的槲皮苷、金丝桃苷进行了鉴别;采用高效液相色谱(HPLC)法对槲皮苷、金丝桃苷进行了含量测定,Diamonsil C18(200 mm×4.6 mm,5 μm,NO.8022889)色谱柱,乙腈-0.1%磷酸(18∶82)为流动相,流速为1.0 ...     

HPLC法测定6种当归植物中紫花前胡素的含量

目的建立了6种当归植物中紫花前胡素的含量测定方法。明确当归属6种植物中紫花前胡素含量，并对其进行比较。方法HPLC法，色谱柱：TIANHE C18（250mm×4．6mm．5μm），流动相H3PO4-NaH2P04缓冲液（稀释10倍）：乙腈（1：1），流速：1mL·min^-1．检测波长：270nm。结果紫花前胡-t＇A0．343175μg·mL^-1之间线性关系良好（r=0．9999），平均回收率：99．3％（n=5），RSD：1．5％。结论6种植物中，每种植物的紫花前胡素含量有差异，且同一种植物里部位不同，前胡素含量也不同。建立的HPLC法简便易行，准确效率高。在实验研究中适用于当归类植物的紫花前胡素的定量测定。     

高效液相色谱-串联质谱法测定羌活中异欧前胡素和紫花前胡苷北大核心CSCD

目的:建立高效液相色谱-串联质谱法测定羌活中异欧前胡素和紫花前胡苷含量。方法:羌活样品超声萃取甲醇稀释后分析。色谱分离采用C18反相色谱柱(150 mm×2.1 mm,3.5μm),流动相为0.1%乙酸水溶液和乙腈,梯度洗脱。串联质谱在多反应监测模式下检测目标分析物,以保留时间和特征离子对(母离子和2个碎片离子)信息比较进行定性和定量分析。结果:异欧前胡素和紫花前胡苷的检出限分别为0.03 ng·mL-1和0.015 ng·mL-1,定量限分别为0.10 ng·mL-1和0.05ng·mL-1,异欧前胡素和紫花前胡苷平均回收率分别为93.6%和94.5%,RSD分别为2.4%和2.6%。结论:该方法经方法学验证,可用于羌活中异欧前胡素和紫花前胡苷的分析。     

紫花前胡苷在大鼠体内的药代动力学研究

目的研究中药狭叶羌活和宽叶羌活主要化学成分之一的紫花前胡苷(ND)在大鼠体内的药代动力学。方法♂SD大鼠(200～220)g随机分组,每组5只,单次尾静脉注射,给药量为80mg.kg-1,从眼眶静脉丛分时取血、处理。采用反相高效液相色谱法、恒速洗脱,应用外标法测定ND在SD系大鼠血浆中的浓度,应用3P87软件计算主要药代动力学参数。结果在所建立的方法学下,ND的保留时间为6.7min;标准曲线为Y=2.0×10-4X+0.4(r=0.9999),在0.2～80mg.L-1的血浆浓度范围内呈良好的线性关系;血浆中最低检测浓度为0.01mg.L-1(S/N=3),最低定量浓度为0.1mg.L-1(S/N=10);在1.0、10.0和40.0mg.L-1低、中、高3个浓度下的提取回收率为76.23～83.72;日内和日间RSD均小于5.60%(n=3)。在室温和冷冻—解冻试验中,ND具有良好的稳定性。按80mg.kg-1单剂量单次静脉给药后,ND在大鼠体内的药代动力学过程符合开放二房室模型,主要药代动力学参数t1/2α、t1/2β、AUC、Vc、Vp、Vss和CL分别为7.02min、219.27min、1384.34mg.min.L-1、0.92L.kg-1、6.04L.kg-1、6.96L.kg-1和0.06L.kg-1.min-1。结论所建立的方法快速、准确、简便,能够满足ND药代动力学研究要求。按80mg.kg-1单剂量单次静脉给药后,ND在大鼠体内分布广泛,消除速度中等。     

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.