HPLC-ELSD法测定桔贝合剂中贝母素甲与贝母素乙的含量

目的建立高效液相色谱-蒸发光散射检测器(HPLC-ELSD)法同时测定桔贝合剂中贝母素甲和贝母素乙含量。方法采用Thermo Hypersil GOLD C_(18)(250 mm×4.6 mm,5μm)色谱柱,乙腈-水-二乙胺(60∶40∶0.03)为流动相,柱温为25℃,流速为1.0 mL·min~(-1);ELSD参数:漂移管温度为85℃,载气流速为2.0 L·min~(-1)。结果贝母素甲在0.572~5.720μg(r=0.9998),贝母素乙在0.795~7.950μg(r=0.9996)与峰面积的对数值呈线性关系;平均加样回收率分别为99.0%(RSD=0.93%)和99.4%(RSD=0.64%)。结论该方法准确可靠、重复性好,为提高桔贝合剂的质量标准提供了科学依据。     

蝉贝合剂中贝母素甲的含量测定

目的运用高效液相色谱-蒸发光散射检测法(HPLC-ELSD)测定蝉贝合剂中有效成分贝母素甲的含量。方法采用Agilent 1200 LC高效液相色谱仪,以甲醇-乙腈-0.5%醋酸溶液(10∶18∶72)为流动相,流速0.7 mL·min-1,Alltech 3300蒸发光散射检测器检测。结果贝母素甲线性范围为0.25～2.50 mg·mL-1,平均回收率为104.6%,RSD=1.5%。结论 HPLC-ELSD测定蝉贝合剂中贝母素甲含量,方法有效,实用。     

高效液相色谱-蒸发光散射器法测定川贝末胶囊中贝母素乙含量

目的建立测定川贝末胶囊中贝母素乙含量的高效液相色谱-蒸发光散射器(HPLC-ELSD)法。方法采用IntersiL C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%二乙胺(70∶30)为流动相,流速为1.0 mL/min,蒸发光散射检测器检测。结果贝母素乙进样质量的线性范围为0.6～6.0μg(r=0.999 5),平均加样回收率为100.16%,RSD为1.69%(n=6)。结论该法简便快速、结果准确可靠,能准确测定川贝末胶囊中贝母素乙的含量,可用于该制剂的质量控制。     

高效液相色谱法分析湖北贝母花中生物碱成分

目的:研究湖北贝母花与鳞茎生物碱成分的区别。方法:根据本实验室建立的湖北贝母指纹图谱,采用高效液相色谱-蒸发光散射检测法,以Hypersil ODS(250mm×4.6mm,5μm)为色谱柱,流动相甲醇(含0.05%三乙胺)-水梯度洗脱,流速为1.0mL.min-1,记录时间60min;以蒸发光散射检测器(ELSD)检测,检测条件漂移管温度75℃,氮气流速1.9L.min-1。结果:湖北贝母花中的生物碱成分得到很好的分离。结论:与湖北贝母指纹图谱对比,并与薄层色谱对照,湖北贝母花与鳞茎部位生物碱成分大体一致,花中浙贝甲素含量较鳞茎高而湖贝甲素含量较鳞茎低,花中湖贝甲素苷的含量很低。     

高效液相色谱-蒸发光散射检测法测定鹤蟾片中贝母素甲和贝母素乙的含量

目的:建立高效液相色谱-蒸发光散射检测法同时测定鹤蟾片中贝母素甲和贝母素乙的含量。方法:采用 HiQsil C_(18)(250 mm×4.6 mm,5 μm)色谱柱,柱温为25℃,以乙腈-甲醇-二乙胺(60:40:0.05)为流动相,流速为0.8 mL·min~(-1);蒸发光散射检测器检测,漂移管温度为80℃,气体流速为2.0 L·min~(-1)。结果:在选定的色谱条件下,以峰面积的常用对数(Y)对进样量的常用对数(X)进行线性回归,贝母甲素和贝母乙素的回归方程分别为 Y=1.557X 1.666和 Y=1.700X 1.571,线性范围分别为0.29～2.80μg(r=0.9996)和0.34～3.40μg(r=0.9999);加样回收率(n=5)分别为99.2%和99.4%,RSD 分别为2.7%和3.1%。结论:本法简便、准确,重复性好,可用于该制剂的质量控制分析。     

HPLC—ELSD法测定盐酸赖氨酸葡萄糖注射液的含量

目的:建立高效液相色谱-蒸发光散射检测法测定盐酸赖氨酸葡萄糖注射液中盐酸赖氨酸及葡萄糖含量。方法:采用 Covasil-C_(18)柱(250 mm×4.6 mm,5μm),以0.05 mol·L~(-1)醋酸铵-甲醇-乙腈(1:1:1)为流动相,流速为0.5 mL·min~(-1)。蒸发光散射检测器(ELSD),漂移管温度为40℃,空气流速为1.5 L·min~(-1)。结果:在选定色谱条件下,盐酸赖氨酸、葡萄糖分别在0.3～0.65 g·L~(-1)(r=0.9997),1.5～3.15 g·L~(-1)(r=0.9994)范围内呈良好的线性关系,回收率分别为98.6%～100.3%,97.8%～99.5%。结论:本法简便,专属性好,可用于盐酸赖氨酸葡萄糖注射液中盐酸赖氨酸及葡萄糖含量检测。     

HPLC-ELSD法测定玉屏风胶囊中黄芪甲苷的含量

目的建立高效液相-蒸发光散射检测法(HPCL-ELSD)测定玉屏风胶囊中黄芪甲苷含量的方法。方法色谱柱为phenomenexGenini(5μm,4.6×250mm,C18);流动相为乙腈-水(75∶25);流速为1.0mL/min,柱温:30℃。蒸发光散射(ELSD)检测条件:漂移管温度为70℃,雾化管温度为40℃。结果黄芪甲苷在0.3038~1.519μg范围内呈良好的线性关系,r=0.9997,平均加样回收率为98.36%,RSD为0.58%(n=6)。结论本法简便快捷,具有良好的精密性和稳定性,测定结果可靠,可用于本产品的质量控制。     

平贝母药材质量控制中适宜测定指标的研究

目的:通过对平贝母中平贝碱甲、贝母素甲、贝母素乙3个单体生物碱成分以及总生物碱测定结果的比较研究,确定可控制平贝母质量的合理测定指标。方法:采用 HPLC—MS 法,色谱柱为 Alltima C_(18)色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.03%二乙胺(30∶70),流速:1 mL·min~(-1);质谱离子源为 ESI;检出模式为正离子检测;电离电压为3.2 kV;电喷雾接口干煤气(N_2)流速为300 L·h~(-1),离子源温度为105℃,锥孔电压为30 V;脱溶剂温度为150℃。选择离子质量数:464.7(平贝碱甲)、432.7(贝母素甲)、430.7(贝母素乙)。测定平贝母中平贝碱甲、贝母素甲、贝母素乙的含量。另以三氯甲烷-甲醇(4∶1)混合溶剂提取总生物碱,以溴麝香草酚蓝比色法在 pH 4.2,紫外吸收波长413 nm 下测定平贝母中总生物碱的含量,结果:LC—MS 法测定平贝母中平贝碱甲、贝母素甲、贝母素乙的含量分别为5～19,1～3,2～3μg·g~(-1);溴麝香草酚蓝比色法测定总生物碱含量以贝母素乙计为600～1000 μg·g~(-1)。结论:平贝母中单-生物碱成分含量太低,不宜作为平贝母药材的质量控制指标,以溴麝香草酚蓝比色法测定总生物碱含量可更客观反映平贝母药用物质情况,更适用于平贝母药材的质量控制。     

HPLC—ELSD法测定盾叶薯蓣中薯蓣皂苷的含量

目的:建立盾叶薯蓣中薯蓣皂苷的含量测定方法。方法:采用高效液相色谱-蒸发光散射检测法(HPLC—ELSD),Allti-ma C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-水(52:48),流速1.0 mL·min~(-1);蒸发光散射检测器参数:漂移管温度40℃,气压3.5 kPa。结果:薯蓣皂苷在0.24～4.80μg(r=0.9995)范围内线性关系良好,平均回收率为97.9%(RSD=1.1%,n=6)。结论:该方法简便、快速、灵敏、准确、重现性好,可作为盾叶薯蓣的质量控制方法。     

HPLC-ELSD测定参贝止咳片中的贝母素甲和贝母素乙

目的 采用HPLC-ELSD测定参贝止咳片中贝母素甲和贝母素乙的含量.方法 采用Lichrospher C18色谱柱(250 mm ×4.6mm,5μm),流动相为乙腈-0.05％二乙胺溶液(70∶30),流速1.0 mL· min-1,柱温30℃,ELSD检测器条件:载气(N2)流速2.5 L·min-1,漂移管温度70.0℃.结果 0.1216～1.4586 μg贝母素甲、0.1005 ～ 1.206 μg贝母素乙进样量对数与峰面积对数具有良好线性关系(r分别为0.9994、0.9995),平均回收率分别为97.22％ (RSD=1.20％)、97.69％(RSD=0.82％).结论 所用方法可靠、准确、重复性好,可用于该制剂的含量测定及质量控制.     

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.     

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.