高效液相色谱法测定那格列奈含量及其有关物质

目的:采用高效液相色谱法(HPLC)测定那格列奈原料、片剂、胶囊的含量及相关物质检查方法.方法:采用Symmetry C18色谱柱(4.6 mm×250 mm,5 μm);以乙腈-磷酸盐溶液(每1 000mL水含磷酸二氢钾4.08 g,用磷酸调节pH 2.8)(41:59)为流动相;流速:1.0 mL·min-1;检测波长:214 nm,柱温:室温;进样量:10μL.结果:在该色谱条件下,那格列奈在49～295 mg·L-1范围内呈良好线性关系,r=0.999 9(n=6),平均回收率为99.5%,RSD为1.4%,检测限为7μg·L-1.结论:该方法简便、准确,可作为那格列奈含量及有关物质的测定方法.     

HPLC法测定缬沙坦胶囊中缬沙坦的含量及有关物质

目的建立缬沙坦胶囊中缬沙坦的含量测定及有关物质的检测方法。方法采用ODS色谱柱(250 mm×4.6 mm,5μm),流动相以[乙腈-水-冰醋酸(40∶60∶0.1)]-[乙腈-水-冰醋酸(70∶30∶0.1)](梯度洗脱),流速1.0 mL/min,柱温30℃,检测波长250 nm。结果主要成分与有关杂质分离效果较好,缬沙坦在19.6～98.0μg/mL(r=0.9992)范围内线性关系良好,加样回收率为99.5%(RSD=0.96%,n=9),缬沙坦杂质Ⅱ的检测限和定量限分别为0.1μg/mL和0.2μg/mL。结论本文建立了HPLC法同时检测缬沙坦的含量及有关物质,方法灵敏,准确,重复性好,可用于缬沙坦胶囊的质量控制。     

那格列奈片的高效液相色谱分析

目的:建立那格列奈片中主药那格列奈及杂质的高效液相色谱测定方法。方法:采用Shim-Pack CLC-ODS(150mm×6.0mm)色谱柱,甲醇-0.02mol.L~(-1)磷酸盐缓冲液(pH6.6)(75:25)作流动相,检测波长210nm,含量测定采用外标法, 杂质检查采用面积归一化法。考察了不同流动相下那格列奈的色谱行为及其与杂质的分离情况。结果:在优化的色谱条件下,片剂辅料不干扰测定,有关杂质与主药那格列奈分离良好,那格列奈线性范围5.1～101.0 mg·L~(-1),最低检测限0.2 mg·L~(-1),含量测定的回收率99.4％～100.4％,RSD<1.7％。结论:此法简便,准确,专属性强,重现性好,可用于那格列奈片的质量控制、稳定性考察和有效期预测。     

反相离子对高效液相色谱法测定左卡尼汀注射液中杂质A的含量

目的:建立离子对高效液相色谱法测定左卡尼汀注射液中杂质A的含量。方法:采用Zorbax SB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为磷酸盐缓冲液[(取磷酸11.5 mL,加水约1800 mL,用1 mol.L-1氢氧化钠溶液调pH至3.0,加水至2000 mL),加庚烷磺酸钠1.1 g,振摇使溶解]-甲醇(95∶5),流速1.0 mL.min-1,检测波长为215 nm。结果:左卡尼汀杂质A与主药峰得到良好分离,分离度达到1.772,在1～100μg·mL-1的范围内,杂质A浓度与峰面积之间呈现良好线性关系,r2=0.9996(n=6),3批左卡尼汀注射液中杂质A的平均含量为0.0052%。结论:该方法简便、准确、专属,可用于该注射液的质量控制。     

RP-HPLC法测定盐酸纳洛酮原料药含量及相关物质

目的:改进盐酸纳洛酮原料药的含量测定和杂质检查方法,为其质量控制提供快速、有效的分析手段。方法:色谱柱为Dikma公司ODS柱(4.6 mm×250 mm,5μm);流动相为水(每580 mL含辛烷磺酸钠1.36 g、氯化钠1.0 g、磷酸1mL)∶甲醇∶乙腈(5.8∶4.2∶1.08);流速0.9 mL/min;柱温40℃;检测波长为229 nm。结果:盐酸纳洛酮在0.100 7~1.611 2μg范围内线性关系良好(r=0.999 9),低、中、高3种浓度(15.0、20.0、25.0μg/mL)对照品溶液的日内RSD分别为0.84%、0.53%、0.67%(n=6),日间RSD分别为0.81%、0.67%、0.46%(n=6),最低检测浓度为50.0 ng/mL。相关杂质、中间体与盐酸纳洛酮分离良好。结论:该法专属性强,结果准确,重现性好,可用作盐酸纳洛酮含量测定和杂质控制的方法。     

牛黄解毒片中黄芩苷和大黄素的含量测定

目的 研究测定牛黄解毒片中黄苓苷和大黄素的含量.方法 采用高效液相色谱法:色谱柱:SB-C18柱(4.6mm×150mm,5μm);流动相:甲醇-0.2％磷酸;检测波长280nm;柱温:25℃:流速:1.0mL·min-1;结果 黄芩苷在5.952～148.8μg·mL-1范围内线性关系良好(r=0.9994),平均回收率97.5％,RSD为1.93％;大黄素在6.048～151.2μg·mL-1范围内线性关系良好(r=0.9970),平均回收率96.6％,RSD为1.72％.结论 该方法灵敏度高,重现性好,可作为该制剂的质量控制方法.     

克霉唑倍他米松乳膏中主药及杂质二苯基-（2-氯苯基）甲醇含量测定方法的改进

目的:建立测定克霉唑倍他米松乳膏中克霉唑和二丙酸倍他米松的含量及其杂质二苯基-(2-氯苯基)甲醇的含量测定方法。方法:采用HPLC法,Waters C18(150 mm×4.6 mm,5μm)色谱柱,甲醇-0.05 mol/L磷酸二氢钾溶液(7∶3)用稀磷酸调节pH至5.75为流动相,流速1.5 mL/min,克霉唑和二丙酸倍他米松检测波长240 nm,杂质检测波长215 nm。结果:克霉唑在20.84～521.00μg/mL范围内线性关系良好(r=0.9997),平均回收率为100.34%,RSD=0.77%(n=9);二丙酸倍他米松在1.29～32.30μg/mL范围内线性关系良好(r=0.9999),平均回收率为100.34%,RSD=0.95%(n=9);杂质二苯基-(2-氯苯基)甲醇在5.096×10-3～0.2038μg/mL范围内线性关系良好(r=0.9999),平均回收率为99.83%,RSD=1.26%(n=9),限度为不得大于克霉唑标示量的2.00%。结论:所建方法准确、简便、快速,适用于克霉唑倍他米松乳膏中克霉唑和二丙酸倍他米松的含量测定和杂质二苯基-(2-氯苯基)甲醇的含量测定。     

HPLC测定岩黄连注射液中岩黄连碱的含量

目的:建立 HPLC 法岩黄连注射液中岩黄连碱的含量。方法:Luna C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(1:1,1000 mL 含磷酸二氢钾3.4 g,十二烷基硫酸钠1.7g),流速1.0 mL·min~(-1),检测波长347 nm,柱温为室温。结果:岩黄连碱在5.48～49.34μg·mL~(-1)浓度范围内呈良好的线性关系(r=1.0000);方法平均加样回收率为99.96%,RSD=0.41%(n=6)。结论:本法可作为岩黄连注射液的质量控制方法。     

苯磺酸氨氯地平原料中基因毒性杂质苯磺酸甲酯与苯磺酸乙酯的含量测定

目的:建立苯磺酸氨氯地平原料中基因毒性杂质苯磺酸甲酯与苯磺酸乙酯含量的高效液相色谱法(HPLC)测定方法。方法:采用Welch Ultimate Plus-C18(Ⅱ)色谱柱(250 mm×4.6 mm,5μm),以磷酸三乙胺缓冲液(pH=3.0)-乙腈为流动相梯度洗脱,检测波长为220 nm,流速为1.0 mL/min,柱温为30℃,研究苯磺酸氨氯地平原料中基因毒性杂质苯磺酸甲酯与苯磺酸乙酯定量测定。结果:苯磺酸甲酯和苯磺酸乙酯吸收峰与氨氯地平能达到有效分离;苯磺酸甲酯浓度在0.102 9～1.633 9μg/mL范围内呈良好线性关系(r=0.999 8),平均回收率为103.6%;苯磺酸乙酯浓度在0.103 3～1.656 1μg/mL范围内呈良好线性关系(r=0.999 8),平均回收率为102.7%;共检测6批苯磺酸氨氯地平原料,且均未检测出苯磺酸甲酯与苯磺酸乙酯的量。结论:本法专属性强,准确度与重复性良好,可用于苯磺酸氨氯地平原料中基因毒性杂质苯磺酸甲酯与苯磺酸乙酯的控制。     

反相高效液相色谱法测定硝酸毛果芸香碱滴眼液的含量和有关物质

目的:建立一种反相高效液相色谱法测定硝酸毛果芸香碱滴眼液的含量和有关物质。方法:用迪马ODS柱(4.6 mm×250 mm,5μm),甲醇-缓冲液(取磷酸13.5 mL,加水700 mL、三乙胺3 mL,加水稀释至1000 mL,用20%氢氧化钠溶液调节pH值至3.00±0.02)(80∶920)为流动相;流速为1.0 mL.min-1;柱温为30℃;UV检测波长220 nm。结果:该色谱条件下,毛果芸香碱与各杂质之间分离良好。毛果芸香碱在2-7μg.mL-1(r=0.9992)、50-350μg.mL-1(r=0.9996)范围内,线性关系良好;含量测定精密度试验RSD=0.50%(n=6);平均回收率为100.5%(n=9);有关物质精密度试验RSD=0.54%(n=6)。结论:本方法简便、快捷、准确、灵敏,可有效控制产品质量。     

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.