RP-HPLC-UV法同时测定兔儿伞中3种黄酮苷类成分的含量

目的建立反相高效液相色谱法同时测定兔儿伞药材中大豆苷、异槲皮苷和槲皮苷共3个黄酮苷类成分的含量。方法采用Hypersil ODS-2色谱柱(250 mm×4.6 mm,5μm),以乙腈-体积分数0.3%磷酸水溶液为流动相,进行梯度洗脱,流速为1.0 m L·min-1,检测波长360 nm,柱温30℃。结果标准曲线线性关系良好(r>0.999 2),精密度,稳定性和重复性实验均符合要求,加样回收率为97.38%~103.30%;对不同产地的兔儿伞药材进行了含量测定,上述3个黄酮苷类成分均能达到基线分离。结论本方法可用于兔儿伞药材的质量控制,同时对于兔儿伞药材的道地性分析也提供了科学依据。     

银杏叶中总黄酮醇苷成分定量分析方法研究

目的:建立银杏叶药材中总黄酮醇苷类成分含量测定方法。方法:分别采用高效液相色谱法(HPLC法)和紫外-可见分光光度法(UV法)测定银杏叶中黄酮类成分的含量,所测得结果进行比较。HPLC法为改良药典方法,采用Phenomenex C_(18)(250mm×4.6mm,5μm)色谱柱,柱温35℃,流动相为0.4%磷酸-甲醇(50:50),流速1.0 mL·min~(-1),检测波长为360nm。结果:HPLC法测定山柰酚、槲皮素和异鼠李素分别在1.20-120 mg·L~(-1),1.50-150 mg·L~(-1),0.68-68mg·L~(-1)浓度范围内,线性关系良好(r=1.000);UV法芦丁在8.84-26.52 mg·L~(-1)浓度范围内,线性关系良好(r=0.9993),加样回收率为102.4%。采用HPLC法测定银杏叶中总黄酮含量(上述3种成分含量之和×2.51)显著低于UV法测定结果,说明银杏叶药材中含有其他酚酸类成分。结论:该方法简便、准确、重现性好。     

胶束电动毛细管色谱法测定山茱萸中莫罗苷和番木鳖苷的含量

目的:建立胶束电动毛细管色谱法测定山茱萸中莫罗苷和番木鳖苷含量。方法:探讨了运行缓冲液中硼酸浓度、胶束(SDS)浓度、有机改性剂(乙腈)比例和 pH 对两组分迁移时间的影响。采用胶束电动毛细管色谱内标法定量。用未涂层石英玻璃毛细管(60 cm×75 m,有效长度45 cm),检测波长240 nm,重力进样(10 cm×5 s),运行电压14 kV,温度20℃,运行缓冲液为0.2 mol·L~(-1)硼酸-0.02 mol·L~(-1)SDS-5%乙腈(0.1 mol·L~(-1)氢氧化钠溶液调节 pH 至10.5)。进样前以0.5 mol·L~(-1)氢氧化钠溶液和运行缓冲液各冲洗3 min。结果:莫罗苷和番木鳖苷分别在16.9～339μg-mL~(-1)(r=0.9994)和20.3～406μg·mL~(-1)(r=0.9998)的范围内呈良好线性相关;莫罗苷和番木鳖苷的平均回收率分别为98.2%(RSD=2.4%)和96.7%(RSD=3.3%)。结论:该方法简便快速、准确,重复性好,为山茱萸药材的质量控制提供了一种新的方法。     

反相高效液相色谱法同时测定金莲花中4种黄酮类化合物的含量

目的:建立用于金莲花药材质量评价的实用有效的含量测定方法。方法:分析柱为 Thermo Hypersil BDS C_(18)柱(4.6 mm×250 mm,5μm),检测波长为340 nm,流速为0.6 mL·min~(-1),以乙腈-水-冰醋酸(14:86:0.05)为流动相等度洗脱反相高效液相色谱法同时测定金莲花药材中荭草苷-2″-O-β-L-半乳糖、荭草苷-2″-O-β-L-阿拉伯糖、荭草苷和牡荆苷4种黄酮类成分的含量。并采用所建立的方法测定了不同品种、部位和来源的金莲花药材中的4种黄酮类成分含量。结果:线性范围分别为47.7～318μg·mL~(-1)(荭草苷-2″-O-β-L-半乳糖)、3.82～255μg·mL~(-1)(荭草苷-2″-O-β-L-阿拉伯糖)、39.0～260μg·mL~(-1)(荭草苷)、7.20～72.0μg·mL~(-1)(牡荆苷);平均加样回收率(n=9)分别为99.8%,102%,100%,101%。不同金莲花药材中4种黄酮类成分含量差异较大。结论:本方法准确可靠,可用于金莲花药材的质量评价和控制。     

自身酶解法测定黄芩药材中黄芩素和汉黄芩素的含量

目的 探讨一种黄芩药材中黄酮苷元类成分黄芩素和汉黄芩素定量测定方法.方法 通过黄芩自身所含的酶在适宜温度条件下将黄芩药材中的黄芩苷、汉黄芩苷等黄酮苷类成分酶解成黄芩素和汉黄芩素等黄酮苷元,再利用高效液相色谱法进行含量测定.结果 黄芩素酶解完全,色谱峰分离良好,且同一流动相下可同时测定黄芩素和汉黄芩素的含量.黄芩素的平均回收率为96.45%,RSD为1.34%(n=9);汉黄芩素的平均回收率为97.86%,RSD为1.89%(n=9).结论 所用测定方法简便、准确,为黄芩素和汉黄芩素的充分提取、分离提供了真实的基础含量数据.     

RP-HPLC法同时测定金莲花中3种碳苷黄酮的含量

目的建立金莲花药材中3种碳苷黄酮含量同时测定的方法,以比较不同产地金莲花药材及不同部位中3种成分的含量。方法采用RP-HPLC法。色谱柱:Shim-pack C18柱(250 mm×4.6 mm,5μm),流动相:乙腈体积分数为0.35%的磷酸溶液(体积比为16∶84),流速:1.0 mL·min-1,检测波长:345 nm,柱温:25℃。结果荭草苷、牡荆苷、荭草素-2″-O-β-L-半乳糖苷能够达到基线分离;荭草苷在9.94～99.40 mg·L-1、牡荆苷在2.49～24.90 mg·L-1、荭草素2″OβL半乳糖苷在10.64～106.40 mg·L-1内线性关系良好。结论本方法为金莲花药材的综合利用和资源开发提供了参考,有利于控制金莲花药材的质量。     

金莲花70%乙醇提取液中黄酮碳苷类成分的研究(Ⅰ)

目的:对金莲花70％乙醇提取物中的黄酮碳苷类成分进行分离和鉴定,为制定金莲花的品质评价标准提供科学数据。方法:用聚酰胺、硅胶、SephadexLH-20色谱柱连续分离和纯化,利用波谱技术对化合物进行结构鉴定。结果:分离得到3个黄酮碳普类化合物,经鉴定分别为荭草素-2″O-β-L-半乳糖苷(2″O-β-L- Galactopyranosyl orientin)、荭草苷(Orientin)和牡荆苷(Vitexin)。结论:首次证明了金莲花中含有荭草素-2″O-β-L -半乳糖苷,并采用聚酰胺-硅胶-SephadexLH-20连续色谱法对该化合物进行了富集。本研究所采用的分离、纯化黄酮碳苷类成分的方法简便、经济、分离效果好。从金莲花中提取黄酮碳苷类成分用70％乙醇为溶剂效果较佳。佥莲花中的抗菌、抗病毒成分主要是黄酮碳苷类化合物,所以,本研究对科学阐释金莲花药效物质基础和有效组分具有重要意义。     

RP—HPLC法同时测定半夏中5种核苷含量的研究

目的:采用反相高效液相色谱法检测半夏药材中的核苷类成分,同时对检测确定的5种核苷进行含量测定。方法:核苷检测采用3种色谱条件,通过与对照品的对照,确定在半夏中5种核苷分别为尿嘧啶、黄嘌呤、尿苷、肌苷、鸟苷。含量测定采用 Ace C_(18)柱(150mm×4.6mm,5μm),以水为流动相,流速:0.8mL·min~(-1),测定波长254nm。结果:尿嘧啶、黄嘌呤为首次在半夏中检测到。尿嘧啶、黄嘌呤、尿苷、肌苷、鸟苷的线性范围依次为0.82～8.16,1.52～15.24,3.30～33.04,0.50～5.02,2.98～29.84μg·mL(-1)。r=0.9995～0.9999;回收率:96.4%～102.3%。结论:以 RP-HPLC 法同时测定半夏药材中的5种核苷快速、简便,结果可靠,为测定半夏中核苷类成分提供了有效的方法。     

毛细管区带电泳分离／安培检测儿茶酚胺药物

目的:建立毛细管区带电泳(CZE)分离/安培检测左旋多巴(DP)、多巴胺(DA)、肾上腺素(EP)、去甲肾上腺素(NE)、5-羟色胺(5-HT)5种儿茶酚胺类神经递质(CAs)药物的方法。方法:采用石英毛细管柱(60 cm×50μm),含8 mmol·L~(-1)对-(季铵盐)杯[4]芳烃(QAC4A)的125 mmol·L~(-1)磷酸氢二钠溶液(Na_2HPO_4,pH 7.0)为缓冲液,分离电压6 kV,分离 CAs 并利用 Cu 微粒修饰碳纤维电极检测。结果:优化条件下5种 CAs 被基线分离,回收率为92%～105%,5种 CAs 在0.2～1000μmol·L~(-1)浓度内呈良好线性关系,最低检测限为0.2 μmol·L~(-1)。应用该方法成功地测定了5种 CAs 注射液、正常人和嗜铬细胞瘤患者尿样。结论:本法用于分离并检测 CAs 药物,具有准确、灵敏、简便的特点,适合该类药品分析。     

高效液相色谱法对淫羊藿及人参复方产品的定性定量分析

目的　建立一种快速有效的分析方法 ,达到鉴定淫羊藿及人参复方产品中黄酮苷类及人参皂苷类成分 ,并测定各成分含量的目的 ,最终控制复方中淫羊藿及人参的比例。方法　通过反相液相色谱 ,对包括药典所收载的淫羊藿药材及其产品进行分析。淫羊藿中的主要黄酮苷类成分 ,以淫羊藿定A、淫羊藿定B、淫羊藿定C、淫羊藿苷为对照品 ;对主要的人参皂苷类成分 ,以人参皂苷Rg1 、Re、Rf、Rb1 、Rb2 、Rd为对照品。结果　在同一色谱条件下 ,对主要黄酮苷类及人参皂苷类成分均可同时测定且分离良好。结论　该方法适用于淫羊藿、人参及其复方产品中淫羊藿黄酮苷类、人参皂苷类成分的定性、定量分析     

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.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

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.     

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.