梯度洗脱高效液相色谱法检测注射用替加环素的有关物质

目的建立测定注射用替加环素有关物质的梯度洗脱反相高效液相色谱法。方法采用Wa-ters C18色谱柱(5μm,416 mm×250 mm),以磷酸盐缓冲液-乙腈(95∶5)为流动相A,以磷酸盐缓冲液-乙腈(50∶50)为流动相B。梯度洗脱条件:0~40 min,A:85%→57%;40~55 min,A:57%→0%;55~56 min,A:0%→85%;以1.0 mL/min的流速进行梯度洗脱,检测波长为248 nm。结果在上述色谱条件下,替加环素与各中间体杂质及降解杂质均能有效分离,分离度大于2.0;检测限为3.6 ng,精密度良好(RSD为0.7%)。结论本方法操作简便,专属性强,灵敏度高,可用于注射用替加环素有关物质的测定。     

HPLC法测定注射用头孢尼西钠含量及有关物质

目的:用 HPLC 法测定注射用头孢尼西钠的含量及其有关物质。方法:采用 Kromasil C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-水-0.2 mol·mL~(-1)磷酸二氢铵(21:74:5)为流动相,流速1.0 mL·min~(-1),检测波长为254 nm;柱温:40℃。结果:头孢尼西钠与其相邻杂质峰能完全分离,头孢尼西钠在22～132μg·mL~(-1)浓度范围内线性关系良好(r=0.9999)。结论:该法简便、准确、专属性好,可以用于注射用头孢尼西钠的含量测定及有关物质检查。     

肾上腺素原料药及盐酸肾上腺素注射液杂质的HPLC-QTOF MS分析

目的:建立HPLC-QTOF MS法测定肾上腺素原料药及盐酸肾上腺素注射液杂质。方法:采用Eclipse plus C18色谱柱(3.0 mm×100 mm,1.8μm),以5 mmol.L-1甲酸铵水溶液-0.1%甲酸水溶液(A)/乙腈(B)为流动相,流速为0.3 mL.min-1,梯度洗脱,梯度条件如下:0→4 min,2%B;4→10 min,2%～50%B;10→12 min,50%～90%B;12→15 min,90%B;柱温为30℃。采用正离子模式采集数据。结果:原料药和注射液中共检出3个杂质,分别为肾上腺酮、肾上腺素磺化物和4-[2-(丁基-2-基氨基)-1-羟乙基]苯酚。结论:本法简便、灵敏,准确,可用于肾上腺素原料药及制剂的杂质定性分析。     

牛胆粉高效液相色谱法特征图谱研究

目的:探求比较黄牛胆粉和水牛胆粉的HPLC特征图谱,为建立牛胆粉的特征图谱提供依据。方法:采用梯度洗脱技术进行HPLC分离分析。色谱柱为Lichrospher C18(4.6 mm×250 mm,5μm),以乙腈-0.001 mol.L-1 KH2PO4溶液为流动相,梯度洗脱(0～20 min,乙腈30%→30%;20～40 min,乙腈30%→40%;40～70 min,乙腈40%→50%;70～80 min,乙腈50%→50%),流速1.0mL.min-1;柱温30℃;DAD检测器,检测波长210 nm。用聚类分析方法处理色谱特征信息,比较色谱以胆酸为参照色谱峰的22个特征峰的信息。结果:通过分析10批黄牛胆粉和水牛胆粉样品,确定了22个共有峰,建立了牛胆粉的HPLC特征图谱。结论:黄牛胆粉和水牛胆粉的色谱指纹特征比较相近,但特征成分含量有一定差异。     

HPLC-CAD同时测定知母中芒果苷和知母皂苷BⅡ的含量

目的:建立HPLC-CAD方法同时测定知母中芒果苷和知母皂苷BⅡ的含量.方法:采用Acclaim-C18色谱柱(150 mm × 4.6 mm,3 μm),以乙腈(A )-0.2％醋酸水溶液(B)为流动相,梯度洗脱(0～10 min,15％A → 21％A;10～12 min,21％A → 23％A,12～30 min...     

HPLC法测定匹伐他汀钙中6种有关物质

目的建立了高效液相色谱梯度洗脱法测定匹伐他汀钙中有关物质的方法。方法采用C18（4.6mm×250mm,5μm）色谱柱,以醋酸盐缓冲液（取醋酸钠0.15g,冰醋酸0.5mL,加水稀释至1000mL）为流动相A,乙腈为流动相B,梯度洗脱条件：0～20min,A为56%;20～40min,A为56%→30%;40～60min,A为30%;以1.0mL.min-1的流速进行梯度洗脱;检测波长为245nm。结果匹伐他汀钙与各有关物质的分离度良好,匹伐他汀钙及杂质Ⅰ、Ⅱ、Ⅲ、Ⅴ、Ⅵ的检测限均为0.04μg.mL-1。结论所建立的方法简便、灵敏、准确,可用于匹伐他汀钙有关物质的测定。     

梯度高效液相色谱法测定苯酰甲硝唑原料药的有关物质

目的:建立梯度高效液相色谱法测定苯酰甲硝唑有关物质的方法。方法:采用梯度洗脱方法。色谱柱为Phen-omsil C18(250mm×4.6mm,5μm);流动相A为磷酸盐缓冲液(取磷酸二氢钾1.5g,加水溶解并稀释至1 000mL,用磷酸调节pH至3.2),流动相B为乙腈,以0.9mL.min-1流速进行梯度洗脱,梯度洗脱条件:0~5min,A为76%;5~15min,A从76%→65%;15~60min,A为65%;60~60.1min,A从65%→76%;60.1~75min,A为76%;检测波长为232nm;柱温40℃;进样量10μL。照杂质对照品对照法和不加校正因子的主成分自身对照法检查。结果:杂质2-甲基-5-硝基咪唑、甲硝唑、苯甲酸均能达到很好的分离,并分别在0.2516~5.032mg.L-1(r=0.999 9),0.2456~4.912mg.L-1(r=1.000 0),0.066 8~1.336mg.L-1(r=0.999 9)范围内具有良好的线性关系。2-甲基-5-硝基咪唑、甲硝唑、苯甲酸的平均回收率(n=9)分别为97.93%(RSD为1.03%),97.63%(RSD为0.46%),...     

HPLC法测定氧化樟脑注射液的有关物质

目的:建立高效液相色谱法测定氧化樟脑注射液的有关物质,并测定2个杂质的校正因子。方法:采用Kromasil 100-5C18(250 mm×4.6 mm,5"m)色谱柱,流动相为95%乙腈溶液(A)-0.2%磷酸溶液(B),线性梯度洗脱[0 min,20%A;0～20min,20%A→80%A,保持15 min],流速1 mL·min-1,检测波长286 nm。结果:样品中氧化樟脑峰与杂质峰分离良好;初步鉴定了2个主要杂质的结构,采用加校正因子的主成分自身对照法控制其质量。结论:所建立的方法简便、快速,可准确地测定氧化樟脑注射液中的有关物质。     

重组人促卵泡激素氧化亚基分析方法研究

目的:研究建立重组人促卵泡激素(rhFSH)氧化亚基分析方法,制备稳定的氧化亚基分析用系统适用性溶液。方法:通过加入过氧化氢氧化破坏重组人促卵泡激素,后加入甲硫氨酸终止该氧化作用,制备系统适用性溶液。采用RP-HPLC法分离分析重组人促卵泡激素氧化亚基。使用C₄色谱柱(250 mm×4.6 mm, 5μm);流动相A为0.2 mol·L^-1磷酸二氢钾缓冲液(pH 2.5),流动相B为80%乙腈;流速1 mL·min^-1;柱温30℃;检测波长210 nm;梯度洗脱(0～10 min, 82%A→74%A;10～25 min, 74%A→60%A;25～26 min, 60%A→20%A;26～56 min, 20%A;56～56.1 min, 20%A→82%A;56.1～75 min, 82%A)。结果:加入甲硫氨酸可有效终止氧化作用,制备得到稳定的系统适用性溶液。优化建立的氧化亚基分析方法专属性良好,重复性(RSD=0.22%)、色谱柱耐用性(RSD=3.21%)、36 h内溶液稳定性(RSD=0.65%)结果良好。9...     

高效液相色谱法检测盐酸厄洛替尼片中有关物质

目的建立检测盐酸厄洛替尼片中有关物质的高效液相色谱法。方法采用资生堂MG C 18色谱柱(250mm×4.6 mm,5μm);流动相A为0.05 mol·L-1磷酸二氢钾溶液(用磷酸调p H至3.0)-甲醇-乙腈(56:22:22),流动相B为甲醇-乙腈-水(45:45:10),梯度洗脱,流速1.0 m L·min-1;检测波长为247 nm。结果在调整后的流动体系下,厄洛替尼峰与相邻杂质峰达到有效分离;样品溶液室温放置12 h,峰面积无明显变化,溶液稳定性较好;各杂质的回收率在99.4%~102.4%。结论该方法准确、可靠,重现性好,可用于该药品的质量控制。     

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.     

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.