高效液相色谱法同时测定人血中对乙酰氨基酚和咖啡因含量

目的:建立同时测定人血浆中对乙酰氨基酚和咖啡因的反相高效液相色谱(RP-HPLC)方法.方法:以茶碱为内标,色谱分离采用Zorbax-Sb-C18分析柱(4.6 mm× 250 mm,5μm),柱温40℃,以乙腈-甲醇-缓冲液(7:7:86,缓冲液内含甲酸0.2%、甲酸铵18 mmol·L-1)为流动相;流速:1.0 mL·min-1,进样量:20μL;紫外检测波长:273 nm.结果:血浆中对乙酰氨基酚和咖啡因测定方法的线性范围分别为0.125～8 mg·L-1,0.0125～0.8 mg·L-1;方法回收率分别为99.76%～105.5%,98.35%～114.45%;日内精密度RSD分别小于3.0%,5.0%;日间精密度RSD分别小于等于12.0%,10.0%.结论:本方法操作简便,选择性好,准确、灵敏,可用于对乙酰氨基酚和咖啡因的人体药动学研究.     

HPLC法测定盐酸多西环素缓释药线中盐酸多西环素的含量

目的 建立盐酸多西环素缓释药线中盐酸多西环素的含量测定方法.方法 采用HPLC法测定药线中盐酸多西环素的含量.Alltima C18(4.6 mm×250 mm, 5 μm)柱,流动相为0.05 mol·L-1草酸铵溶液∶N,N-二甲基甲酰胺∶0.2 mol·L-1磷酸氢二铵溶液(65∶30∶5),用氨试液调节pH值为8.0±0.2,检测波长280 nm,流速为1.0 ml·min-1.结果 盐酸多西环素在49～490 mg·L-1范围内线性关系良好,r=0.9997,A=43767C 562001;平均回收率为98.72%,RSD＝1.31%(n=6 ).结论 该法简单可行,结果准确,可用于该制剂的质量标准检测.     

高效液相色谱法测定人体血浆及尿液中多西环素的药物浓度

目的:建市人体血浆及尿液中多西环素药物浓度的高效液相色谱分析方法。方法:美他环素作为内标,待测血浆和尿液分别以高氯酸和甲醇沉淀蛋白,离心后取上清液进行高效液相色谱紫外检测,色谱柱为 Kromasil C_(18)柱(5μm,4.6mm×250mm);流动相为乙腈-水(30:70,含5 mmol·L~(-1)的柠檬酸),流速1.0 mL·min~(-1),检测波长350 nm。结果:本测定方法血浆和尿液的线性范围分别为O.1～6.13 mg·L~(-1)和0.085～13.62 mg·L~(-1),相关系数分别为0.9995和0.9991,血浆及尿液的相对回收率分别为95.2%～101.4%,95.6%～105.5%;日内日间 RSD 均小于10%,最低检测限分别为0.026 mg·L~(-1)及0.034mg·L~(-1)。结论:高效液相色谱法测定血浆与尿液中药物浓度简单、快速、可靠,可用于多西环素的药代动力学及生物利用度研究。     

HPLC法监测人血清中茶碱类药物浓度

目的建立同时测定茶碱类药物血药浓度的高效液相色谱法。方法筛选得到的最佳色谱条件为:采用色谱柱Ultimate XB-C18柱(4.6×200mm,5μm),以咖啡因为内标,甲醇-乙腈-水(20∶3∶77)为流动相,流速1.0m L·min-1,检测波长273nm,柱温30℃。采用先沉淀后提取的方法进样测定。结果多索茶碱和茶碱分别在1.5~24mg·L-1(r=1.0000)和2.5~40mg·L-1(r=0.9996)范围内线性关系良好。多索茶碱的低、中、高浓度平均回收率为100.07±5.30%,日间精密度和日内精密度RSD均小于6.5%。茶碱的低、中、高浓度回收率为101.72±4.92%,日间精密度和日内精密度RSD均小于5.5%。结论该方法灵敏度高、准确、重复性好,适用于多索茶碱和茶碱血药浓度的测定。     

高效液相色谱法测定兔房水中丹参素的含量

目的:建立高效液相色谱测定兔房水中丹参素含量的方法,了解兔眼丹参注射液直流电离子导入后房水中丹参素的吸收情况.方法:色谱条件:色谱柱为Hypersil Division ODS C18分析柱(150 mm×4.6 mm,5 μm);流动相为水-甲醇(95∶5),1000 mL中含十二烷基磺酸钠1 g,磷酸二氢钠0.6 g,冰醋酸2 mL;流速为1.0 mL·min-1;检测波长为281 nm;柱温为35℃.结果:在0.52～8.32 mg·L-1范围内,峰面积与浓度呈良好线性关系,r=0.9998,最低检测浓度为0.05 mg·L-1;平均回收率为93.42%;日内和日间RSD均＜10%.结论:建立了特异,简便,灵敏的HPLC检测法,为进一步研究探讨丹参素在眼内作用的机理打下基础.     

高效液相色谱法测定盐酸吡格列酮的血药浓度

目的:建立高效液相色谱法测定人血浆中盐酸吡格列酮浓度的方法.方法:以卡马西平为内标,血浆样品用醋酸乙酯萃取,采用Diamonsil(TM)C18柱(250 mm×4.6mm,5μm)分析.流动相为乙腈-0.05 mol·L-1磷酸二氢钾(pH 6.5)(42:58),流速1.2 mL·min-1,检测波长229 nm,柱温35℃.结果:本法在0.025～4.0 mg·L-1间线性关系良好,γ=0.999 9,RSD为2.08%,(n=6),最低检测质量浓度为0.02 mg·L-1.日内、日间精密度RSD均小于10%,低、中、高浓度的提取回收率均大于99%(n=5).结论:此方法灵敏度高、重复性好,尤其适用于临床治疗合并用药时的药物浓度监测.     

LC-MS/MS法测定人血浆中紫杉醇的浓度

目的建立一种液相色谱-串联质谱(LC-MS/MS)的方法,测定人血浆中紫杉醇的药物浓度。方法色谱柱:Diamonsil C18柱(50 mm×2.1 mm I.D,粒径5μm,北京迪马公司),流动相:乙腈-水-甲酸(体积比为55.0∶45.0∶0.1),采用沉淀蛋白法,以多反应离子监测(multiple reaction monitoring,MRM)扫描方式进行检测,测定肿瘤患者静脉滴注紫杉醇注射液后血浆中的药物浓度。结果血浆中紫杉醇的药物浓度在20.0⁵ 000.0μg·L-1以内时,其线性关系良好,相关系数r=0.995 4;日内和日间精密度RSD≤13.4%;紫杉醇的平均提取回收率为93.6%5 000.0μg·L-1以内时,其线性关系良好,相关系数r=0.995 4;日内和日间精密度RSD≤13.4%;紫杉醇的平均提取回收率为93.6%¹08.5%,紫杉醇的基质效应为102.4%108.5%,紫杉醇的基质效应为102.4%¹05.4%,紫杉醇在人血浆中主要药动学参数如下:t1/2为(5.1±3.0)h,ρmax为(3.8±0.5)mg·L-1,AUC0-36为(13.2±2.6)mg·h·L-1,AUC0-∞为(13.3±2.8)mg·h·L-1。结论该方法适用于紫杉醇在人体内的药物动力学研究。     

高效液相色谱法测定人血清和置换液中美罗培南浓度

目的:建立高效液相色谱法(HPLC)测定人血清和置换液中美罗培南浓度的方法。方法:采用HPLC法,以依利特Hypersil ODS2柱(4.6mm×150mm,5μm)为色谱柱;甲醇-0.01mol·L-1磷酸二氢钾缓冲液(9∶91)为流动相,pH4.91;流速为1.4mL·min-1;检测波长为297nm。结果:血清中美罗培南的标准曲线方程为Y=17.027X-4.7541,r=0.9997,在0.5~80mg·L-1范围内线性关系良好,平均回收率为100.2%,日内,日间RSD均小于7.2%;置换液中美罗培南的标准曲线方程为Y=18.931X 0.8928,r=0.9998,在0.5~80mg·L-1范围内线性关系良好,平均回收率99.2%,日内、日间RSD均小于7.6%。结论:该方法灵敏准确,适用于临床药动学的研究。     

高效液相色谱法测定血中胺碘酮及其活性代谢物浓度

目的:建立反相高效液相色谱测定胺碘酮及其活性代谢物去乙基胺碘酮血药浓度的方法.方法:依利特分析柱HypersilODS2(4.6 mm×250mm,5μm),流动相为乙腈-100 mmol·L-1醋酸(内含15 mmol·L-1二乙胺)(55:45),检测波长242 nm,流速为1.0mL·min,柱温为室温.结果:线性范围0.1～4.8 mg·L,方法回收率90.3%～101.9%,日内RSD小于7%,日间RSD小于11%.结论:适于胺碘酮及其活性代谢物去乙基胺碘酮的临床监测.     

HPLC法测定奥美拉唑镁原料药中醋酸的残留量

目的建立高效液相色谱法测定奥美拉唑镁原料药中有机溶剂醋酸含量。方法以色谱柱:以XBridge TM C18(4.6 mm×250 mm,5μm)为分离柱,采用等度洗脱,检测波长:210 nm。结果线性关系良好,方法精密度,重复性均符合要求。冰醋酸检测浓度线性范围为0.01~0.06 mg·m L-1(r=0.999 994),低、中、高浓度平均回收率(n=9)分别为100.60%、100.38%、100.23%,RSD分别为2.20%、2.21%、1.60%,最低检测限为0.005 0 mg·m L-1。结论该方法可用于奥美拉唑镁原料药中残留溶剂醋酸含量的测定,限度为不得过0.05%(500 ppm)。     

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.     

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.