HPLC法同时测定人血浆中苦参碱和氧化苦参碱含量

目的 :建立测定苦参碱和氧化苦参碱血药浓度的HPLC法。方法 :血浆标本经碱化以氯仿 -正丁醇 (98∶2)提取 ,中性氧化铝柱净化 ,吹干后以乙腈 -无水乙醇 -磷酸水溶液 (80∶10∶8)为流动相 ,槐定碱为内标 ,经LichrosorbNH2 柱分离后在紫外波长220nm处检测。结果 :苦参碱和氧化苦参碱的血药浓度线性范围在1 25～40mg/L ,最低检测限为0 1mg/L(S/N=2)。苦参碱和氧化苦参碱高、中、低3种浓度的平均回收率分别为106 96 %和105 04 % ,日内、日间精密度分别小于13 %和7 %。结论 :此法为苦参碱和氧化苦参碱的血药浓度测定提供了一种简便可行的方法。     

HPLC法同时测定人血浆中甘草酸和苦参碱的质量浓度

目的建立以HPLC法同时测定人血浆中甘草酸和苦参碱质量浓度的方法。方法 Ultimate AQ-C18柱(250mm×4.6mm,5μm),柱温为室温(25℃),流动相为乙腈-三乙胺调pH至7.5的0.05mol·L-1磷酸二氢钾(22.5∶77.5),流速为1.0mL·min-1,检测波长为220nm(检测苦参碱)和250nm(检测甘草酸),进样量50μL。结果甘草酸单铵盐质量浓度在2.0²00.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0200.0μg·mL-1范围内线性良好,苦参碱质量浓度在2.0²00.0μg·mL-1范围内线性良好。平均回收率均大于95%,日内、日间RSD均小于4%。结论该方法简便、快速、灵敏、可靠,可用于人血浆中甘草酸和苦参碱质量浓度的监测。     

心律宁片中苦参碱和氧化苦参碱的含量测定

目的:建立心律宁片中苦参碱和氧化苦参碱含量的测定方法。方法:色谱柱为Alltima Amino(250mm×4.6mm,5μm),流动相为乙腈-无水乙醇-3%磷酸溶液(80:10:10),检测波长为220nm,流速为1.0ml·min~(-1);柱温:室温。结果:苦参碱线性范围是0.168μg～3.360μg,r=0.999 7,氧化苦参碱线性范围是0.124μg～2.480μg,r=0.999 3,苦参碱及氧化苦参碱的平均回收率分别为101.1%(RSD=1.22%)和100.2%(RSD=1.91%)(n=6)。结论:方法快速简便,重复性良好,可用于测定心律宁片中苦参碱和氧化苦参碱的含量。     

HPLC法测定犬血浆中的苦参素和苦参碱

目的:建立同时测定Beagle犬血浆中苦参素和苦参碱的高效液相色谱(HPLC)方法。方法:以高氯酸沉淀蛋白,在强碱条件下用二氯甲烷萃取苦参素和苦参碱,通过HPLC法分别测定苦参素和苦参碱的血药浓度。结果:苦参素和苦参碱分别在0.2～15μg.mL-1和0.1～5μg.mL-1的浓度范围内呈现良好的线性关系,最低检测限均为20ng.mL-1,提取回收率、方法回收率、日内精密度和日间精密度均符合要求。结论:本方法简便、准确、干扰少,灵敏度高,可以同时检测到苦参素和苦参碱,适用于苦参素体内药动学的研究及其体内转化过程和代谢途径的研究,并对研制苦参素的新制剂有一定的指导作用。     

LC-MS法检测人血浆中双氯芬酸的浓度

目的:建立LC-MS测定人血浆中双氯芬酸浓度的方法。方法:待测血浆样品经乙酸乙酯萃取后,用HPLC-MS法进行检测,以格列吡嗪为内标。色谱柱为Agilent ZORBAX SB-C18(150 mm×4.6 mm,5μm),流动相为10 mmol.L-1醋酸铵溶液-甲醇(35∶65),流速为0.5 mL.min-1;质谱采用电喷雾离子化(ESI)方式和选择性离子检测(SIM)模式,检测离子为负离子。结果:本方法线性范围为3.936 0~492.000 0 ng.mL-1,最低定量浓度3.936 0 ng.mL-1,准确度、精密度以及稳定性均符合有关要求。结论:该方法简便、准确,灵敏度高,专属性强,适用于人血浆中双氯芬酸浓度的测定。     

氧化苦参碱在犬体内的药动学

目的:建立beagle犬血浆中氧化苦参碱的LC-MS测定法,测定其灌胃后在beagle犬体内的药动学.方法:采用LichrospherC18柱(250 mm×4.6 mm,5μm),柱温:25℃.流动相为10 mmol·L-1醋酸铵水溶液-甲醇(25:75),流速:1 mL·min-1;电喷雾离子化(ESI)方式,采用选择性离子检测法,检测离子为正离子,氧化苦参碱的离子为[m/z H] 265.1,内标非那雄胺的离子是[m/z H] 373.3.结果:氧化苦参碱在2～5000μg·L-1的范围内呈良好的线性关系(r=0.9990),最低检出限达0.6μg·L-1,日内和日间误差均小于4.2%,萃取回收率大于90%;氧化苦参碱的药-时数据符合二室模型,Cmax为(2 418.3±970.8)μg·L-1,tmax为(1.0±0.3)h,t1/2β为(5.5±1.6)h,AUC0→∝为(6117.4±1080.2)μg·L-1·h.结论:该法灵敏、简单,专属性强,可用于犬血浆中氧化苦参碱的测定.     

HPLC法测定苦参总碱中苦参碱与氧化苦参碱的含量

目的:采用高效液相色谱法测定苦参总碱中苦参碱和氧化苦参碱的含量。方法:采用 Elite Hypersil NH_2柱(5μm,250mm×4.6 mm),乙腈-无水乙醇-3%磷酸溶液(81:10:9)为流动相,流速:1.0 mL·min~(-1),检测波长220 nm,柱温:25℃,进样量:10 μL。结果:苦参碱、氧化苦参碱的线性范围分别为5.32～53.2μg·mL~(-1)(r=0.9997),65.7～525.6μg·mL~(-1)(r=0.9995)。苦参碱平均加样回收率为99.5%,RSD=1.0%(n=6);氧化苦参碱的平均加样回收率为99.7%,RSD=0.6%(n=6)。结论:本方法操作简便、快速、可靠,可用于控制苦参总碱质量。     

LC-MS/MS法测定Beagle犬血浆中右旋酮洛芬的浓度

目的:建立LC-MS/MS法测定Beagle犬血浆中右旋酮洛芬的浓度。方法:血浆样品采用乙腈沉淀法处理,取上清液进样。以乙腈(每500 mL中加入甲酸10μL)-水(80∶20,v/v)为流动相;用Zorbax Eclipse SB-C18柱(5μm,150 mm×4.6 mm)色谱柱分离;以布洛芬为内标。流速0.5 mL·min-1;进样量25μL;柱温为室温。质谱检测采用ESI正离子模式,扫描方式为多反应监测,扫描离子对m/z右旋酮洛芬253.0/209.0,布洛芬204.8/160.8。结果:右旋酮洛芬的线性范围为1.0～1000 ng·mL-1,线性关系良好(r=0.9979),最低定量下限为1.0 ng·mL-1,绝对回收率在100.0%～104.3%之间,变异系数均小于6%。结论:该方法预处理快速简单,灵敏度高,所建立的LC-MS/MS法准确、可靠,可用于犬血浆中右旋酮洛芬的浓度测定及药代动力学研究。     

LC-MS/MS法测定人体利多卡因浓度及药代动力学的应用

目的:建立LC-MS/MS法测定人体血浆利多卡因的浓度,并研究NAL1282利多卡因贴剂(5%)的药代动力学。方法:色谱柱为YMC C18(150 mm×2.1 mm,5μm),流动相为乙腈-20 mmol·L-1甲酸铵(87:13,v/v);流速:0.25 mL·min-1;进样量:5μL;柱温:40℃,样品室温度为15℃。采用随机、单剂量给药、开放性、两周期交叉设计的单中心试验,分别给予9名健康受试者NAL1282利多卡因贴剂3贴,700 mg·贴-1。采用LC-MS/MS法测定给药后不同时间的血药浓度。结果:利多卡因线性范围为0.8～206 ng·mL-1,最低检测限为0.2 ng·mL-1,方法灵敏、稳定、特异性高。利多卡因贴剂主要药代动力学参数Cmax、Tmax、AUC0→24和AUC0→∞分别为(61.6±18.5)ng·mL-1;9 h(6 h,12 h);(848.4±231.7)ng·h·mL-1;(1129.3±324.0)ng·h·mL-1。结论:该方法简便、准确,重复性好,并可应用到人体利多卡因贴剂药代动力学的研究。     

国产与进口复方甘草酸苷片在健康人体的生物等效性

目的 建立快速、灵敏的液相色谱-串联质谱法(LC-MS/MS)测定人血浆中甘草次酸(治疗慢性肝病药物)的浓度,评价国产与进口复方甘草酸苷片在健康人体的生物等效性.方法 22名中国健康男性志愿者随机分为试验组和对照组,采用双交叉设计和单剂量口服方式,以LC-MS/MS法测定血浆中甘草次酸浓度,经DAS 2.0统计软件计算主要药代动力学参数.结果 线性范围0.50～800.00 ng·mL-1,最低定量浓度为0.50 ng·mL-1,日内精密度(RSD)<10.60%,日间精密度(RSD)<8.80%.受试制剂或参比制剂的主要药动学参数:t1/2分别为(13.1±8.5)和(10.3±5.6)h;tmax分别为(13.5±6.0)和(16.7±7.2)h;Cmax分别为(288.0±249.0)和(287.0±160.0)ng·mL-1.试验制剂相对生物利用度为(117.4±79.8)%.结论 试验制剂与参比制剂具有人体生物等效性.     

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.     

Pharmacological treatment of COVID-19: an opinion paper

The precocity and efficacy of the vaccines developed so far against COVID-19 has been the most significant and saving advance against the pandemic. The development of vaccines has not prevented, during the whole period of the pandemic, the constant search for therapeutic medicines, both among existing drugs with different indications and in the development of new drugs. The Scientific Committee of the COVID-19 of the Illustrious College of Physicians of Madrid wanted to offer an early, simplified and critical approach to these new drugs, to new developments in immunotherapy and to what has been learned from the immune response modulators already known and which have proven effective against the virus, in order to help understand the current situation.     

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.