HPLC法测定杞菊地黄丸中莫诺苷、马钱苷和丹皮酚的含量

目的:建立一种快速、准确和实用的HPLC方法,用于同时测定杞菊地黄丸中莫诺苷、马钱苷和丹皮酚的含量。方法:采用Phenomenex Gemini C18(4.6 mm×250 mm,5μm)色谱柱,以乙腈-0.3%磷酸水溶液为流动相梯度洗脱,流速1.0 m L·min-1,柱温40℃,检测波长为240 nm(莫诺苷、马钱苷)和274 nm(丹皮酚)。结果:莫诺苷、马钱苷、丹皮酚质量浓度分别在2.242~44.84μg·m L-1(r=0.999 9)、2.130~42.60μg·m L-1(r=1.000)、5.068~101.4μg·m L-1(r=1.000)范围内,与峰面积线性关系良好;平均回收率分别为96.2%(RSD=0.44%)、96.0%(RSD=0.21%)、103.0%(RSD=0.65%)。结论:本方法可作为杞菊地黄丸中莫诺苷、马钱苷和丹皮酚含量同时测定的方法,适用于杞菊地黄丸的质量控制。     

麝香草酚电化学检测方法的研究

目的建立麝香草酚的电化学检测方法,并用于麝香草酚的含量测定。方法循环伏安法（CV）、差分脉冲伏安法（DPV）。结果在0.10mol.L-1的Na2HPO4-NaH2PO4（pH=9.0）缓冲溶液中,0.050V/S的扫描速度下,麝香草酚的峰电流与其浓度在4.0×10-5～1.6×10-4mol.L-1的范围内呈良好的线性关系,检出限为1.6×10-4mol.L-1。结论该方法准确、可靠,操作简便、快速,可以作为麝香草酚含量测定方法。     

高分辨气相色谱法测定六味地黄丸中丹皮酚的含量

本文建立了中成药六味地黄丸中丹皮酚含量测定的高分辨气相色谱法。色谱条件为SE-30 FSOT柱(L28m×φ0.28mm),柱温180℃,火焰离子化检测器,以正十六烷为内标,六味地黄丸经乙酸乙酯提取,浓缩后测定丹皮酚的含量。在2—12.5μg 范围内,丹皮酚与内标峰面积之比呈线性关系,丹皮酚加样平均回收率为100.3%(CV=1.38%,n=11)。本法可用于六味地黄丸三种制剂的含量测定,测得结果与其它方法比较一致,准确可靠。     

高分辨气相色谱法测定六味地黄丸中丹皮酚的含量

本文建立了中成药六味地黄丸中丹皮酚含量测定的高分辨气相色谱法。色谱条件为 SE-30 FSOT柱(L28m×φ0.28mm),柱温180℃,火焰离子化检测器,以正十六烷为内标,六味地黄丸经乙酸乙酯提取,浓缩后测定丹皮酚的含量。在2—12.5μg 范围内,丹皮酚与内标峰面积之比呈线性关系,丹皮酚加样平均回收率为100.3%(CV=1.38%,n=11)。本法可用于六味地黄丸三种制剂的含量测定,测得结果与其它方法比较一致,准确可靠。     

六味地黄丸(浓缩丸)质量标准研究

目的:建立六味地黄丸(浓缩丸)的质量标准。方法:采用薄层色谱(TLC)法对方中熟地黄、山药、茯苓、泽泻进行定性鉴别;采用高效液相色谱法测定制剂中丹皮酚和马钱苷的含量。结果:TLC的斑点清晰、分离度较好,阴性无干扰;丹皮酚和马钱苷的进样量分别在0.1006～1.006μg(r=0.9999)、0.3096～2.322μg(r=0.9996)范围内与各自峰面积积分值呈良好的线性关系;平均回收率分别为98.2%、95.3%,RSD分别为2.3%(n=6)、0.42%(n=6)。结论:所建标准可用于六味地黄丸(浓缩丸)的质量控制。     

RP-HPLC法测定浓缩六味地黄丸中马钱苷的含量

建立浓缩六味地黄丸中马钱苷的含量测定方法。采用高效液相色谱法,以Hypersil ODS2为色谱柱;流动相:0.05mol.L-1磷酸二氢钠-乙腈(85:15);检测波长:238nm。马钱苷在0.0285～5.2250μg范围内,浓度与峰面积呈良好的线性关系(r=0.9998);回收率:98.39%,RSD=1.2%(n=9)。此方法简便、快速,可作为浓缩六味地黄丸中马钱苷的含量测定方法。     

HPLC法测定知柏地黄丸浓缩丸中丹皮酚的含量

目的 建立知柏地黄丸浓缩丸中丹皮酚的含量测定方法.方法 采用高效液相色谱法,Kromasil KR100-5C18(250×4.6mm)色谱柱,流动相:甲醇-水(70∶30),流速:1.0mL·min-1,检测波长:274nm.结果 丹皮酚在8.168～163.36μg·mL-1范围内线性关系良好.(r=0.9999,n=6),平均回收率99.12%,RSD=0.52%(n=6).结论 本法快速,简便,准确,重复性好,可作为该丸剂的含量测定方法.     

胶束电动毛细管色谱法测定牡丹皮及六味地黄丸中丹皮酚的含量

目的用胶束电动毛细管色谱法分离测定中药材牡丹皮及中成药六味地黄丸中丹皮酚的含量。方法定量采用内标法 ,选取芦丁为内标。所用毛细管规格为 5 5cm(有效长度 5 0cm)× 75 μm,检测波长 2 74nm ,电压 1 5kV ,背景电解质分别为 30mmol/LSDS -30mmol/L硼砂 ,2 0mmol/LSDS -5 0mmol/L硼砂 ( pH 9 4 )。 结果线性分别为 6～ 4 2mg/L,1 8 2～ 91mg/L;相关系数分别为0 9995和 0 9997;RSD为 1 95 %和 3 6 %;加样回收率为 1 0 1 5 %～ 1 0 3 3%和 97 7%～1 0 1 9%。丹皮酚含量分别为 1 5 9%和 0 1 4 8%。结论胶束电动毛细管色谱法用于测定牡丹皮及六味地黄丸中丹皮酚的含量是可行的。     

多巴酚丁胺在磷钨酸铈/碳纳米管复合物修饰电极上的电化学行为及其测定

目的:制备出磷钨酸铈纳米晶负载的碳纳米管修饰电极,研究多巴酚丁胺(DBTM)在该修饰电极上的电化学行为,建立DBTM含量测定的示差脉冲伏安新方法。方法:用浸渍法制备负载磷钨酸铈(CePW)复合材料的CePW/LDWNT修饰电极,用电化学阻抗(EIS)对修饰电极进行表征并考察多巴酚丁胺在不同电极体系中的循环伏安特性。结果:制备的纳米复合物对DBTM具有催化氧化作用,DBTM的氧化峰电流与浓度在2.0×10-6～8.0×10-6mol·L-1范围内呈线性关系,线性回归方程为ip=-10.06+5.824C,r=0.9966,检测限为5.8×10-9mol·L-1(S/N=3)。结论:该法灵敏度高,选择性及重现性好,可用于盐酸多巴酚丁胺注射液中多巴酚丁胺的测定。     

秋水仙碱体外抗乙型肝炎病毒的作用

目的:探讨秋水仙碱在体外抗乙型肝炎病毒(HBV)的作用。方法:以秋水仙碱作用于HBV全基因组转染的人肝癌细胞株(Hep G22.2.15),用罗氏电化学发光免疫分析仪检测HBsAg和HBeAg,用ABI7000荧光定量PCR扩增仪检测细胞内、外HBV-DNA、HBV-共价闭环DNA(HBV-cccDNA)(copies/cell)。数据表示为l g(x珚±s)。结果:(1)秋水仙碱6.25×10-6～1×10-4 mol·L-1能显著抑制Hep G22.2.15产生/分泌HBsAg(P<0.05,P<0.01),IC50=1.473×10-7 mol·L-1,其3.125×10-6和2.5×10-5～1×10-4 mol·L-1能显著抑制Hep G22.2.15产生/分泌HBeAg(P<0.05,P<0.01),IC50=2.387×10-7 mol·L-1,拉米夫定(lamivudine)IC50分别为3.52×10-4 mol·L-1和3.184×10-3 mol·L-1。(2)秋水仙碱和拉米夫定有类似的降低Hep G22.2.15细胞内HBV-DNA的作用(P<0.05),IC50分别为9.747×10-9 mol·L-1和2.649×10-6 mol·L-1。(3)秋水仙碱3.125×10-6和6.25×10-6 mol·L-1能明显减少Hep G22.2.15细胞内和上清液中HBV-cccDNA(P<0.05)。(4)秋水仙碱1.6×10-6～1×10-3 mol·L-1和拉米夫定2×10-4,1×10-3 mol·L-1皆能显著抑制Hep G22.2.15细胞增殖(P<0.05,P<0.01),IC50分别为7.933×10-6 mol·L-1和2.60×10-4 mol·L-1。结论:秋水仙碱在体外具有抗HBV的作用,包括降低HBsAg和HBeAg的产生/分泌,减低Hep G22.2.15细胞内、外HBV-cccDNA水平,减少细胞内HBV-DNA,且主要是由于直接抑制HBV-DNA和HBV-cccDNA的产生。     

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.