HPLC-MS测定人血浆中美利曲辛及其在生物等效性研究中的应用

目的 建立测定人血浆中美利曲辛浓度的高效液相色谱-质谱联用法,并用于氟哌噻吨美利曲辛片的生物等效性研究.方法 血浆样品用乙酸乙酯萃取,采用Thermo Hypersil-HyPURITY C13色谱柱(150 mm×2.1mm,5μm)分离,以50 mmol·L-1乙酸铵溶液(pH=4.0)-甲醇-乙腈(45∶36∶19)为流动相,流速为0.22 mL·min-1.采用高效液相色谱-质谱电喷雾电离法,选择离子监测(SIM).结果 美利曲辛在0.4～50.0 μg·L-1线性关系良好,平均回收率为98.6％,日内RSD≤7.8％,日间RSD≤6.0％.结论 该方法准确、灵敏度高,适用于含美利曲辛制剂的人体药动学和生物等效性研究.     

液相色谱—电喷雾质谱联用测定人血浆中盐酸乙哌立松的浓度

目的:建立测定人血浆中盐酸乙哌立松的高效液相色谱-电喷雾质谱联用(HPLC/ESI-MS)方法。方法:血浆经碱化后,叔丁基甲醚提取,HPLC/ESI-MS选择离子检测。结果:本法线性范围为32～0.25μg·L~(-1),最低检测浓度为0.0μg·L~(-1)(S/N≥3)。血浆中乙哌立松的回收率为99.10～99.85％,日内RSD≤5.9％,日间RSD≥10.8％。结论:本法适用于盐酸乙哌立松的生物等效性研究和血药浓度监测。     

HPLC-MS/MS法快速测定人血浆中氟哌噻吨浓度及生物等效性研究

目的 建立测定人血浆中氟哌噻吨浓度的高效液相串联质谱法(HPLC-MS/MS),用于氟哌噻吨美利曲辛片的生物等效性研究.方法 以Agilent ZORBAX Eclipse Plus C18(4.6mm×150mm,5μm)为色谱柱,流动相为乙腈(含1%甲酸):0.02mol/L甲酸铵水溶液(80:20,V:V),流速:0.8ml/min;柱温:40℃,以醋酸乙酯:二氯甲烷(4:1,V:V)为提取剂.样品经电喷雾离子源正离子化后.通过三重四级杆串联质谱仪.采用选择性反应监测(SRM)对氟哌噻吨(m/z 435.2→305.1)和氯普噻吨(m/z 316.2→231.1)进行测定.结果 氟哌噻吨的高(1μg/L)、中(0.5μg/L)、低(0.05μg/L)3个浓度的平均回收率分别为101.01%、96.19%和106.63%,日内(n=5)、日间(n=3)RSD均小于15%;分析方法的最低定量限为0.025μg/L.线性范围为:0.025～2.5μg/L,回归方程为:F=0.6625ρ+0.0049,r=0.997(n=7),权重为1/ρ2.结论 该方法灵敏、准确、简单、快速,可用于临床血浓监测和药动学研究.     

高效液相色谱法测定人血浆中米卡芬净钠浓度

目的建立高效液相色谱测定人体血浆中米卡芬净钠(抗真菌药)浓度的方法。方法血浆样品经甲醇沉淀蛋白提取分离,色谱柱为CAPCELL PAK C1柱,流动相为甲醇-10 mmol.L-1醋酸铵(57∶43,v/v),流速为0.2 mL.min-1。结果米卡芬净钠在0.2～25.0μg.mL-1线性良好,最低定量限为0.2μg.mL-1,日内RSD均≤4.1%,日间RSD均≤5.0%,提取回收率RSD均≤6.8%。结论本方法灵敏、快速、准确,可用于米卡芬净钠药代动力学测定。     

HPLC法测定大鼠血浆中抗肿瘤化合物SYUIQ-5的浓度

目的建立一种灵敏度较高的高效液相色谱法,测定大鼠血浆中抗肿瘤化合物SYUIQ-5的浓度。方法流动相用35mmol·L-1磷酸氢二钾缓冲液(pH8.0)-甲醇-三乙胺(30∶70∶0.05),血浆样本用乙醚萃取,紫外检测波长为278nm。结果线性范围为30~1500μg·L-1,线性关系良好,最低检测限为20μg·L-1,日内RSD≤5.2%,日间RSD≤3.8%;提取回收率为72.1%~80.5%(n=5),相对回收率为96.8%~105.1%。结论本方法灵敏度高,重现性好,精密度和准确性好,适于SYUIQ-5的药代动力学研究。     

反相高效液相色谱法测定血浆中吗氯贝胺浓度

目的:建立人血浆中吗氯贝胺的反相高效液相色谱检测方法.方法:血浆样品在碱性条件下(pH 11.0)用二氯甲烷提取处理.色谱柱为μ-BondapakTM C18 (3.9 mm×15 0mm,10 μm),柱温37.0 ℃,流动相为乙腈-0.067 mol·L-1磷酸二氢钾溶液(1∶5,pH 2.6),流速1.0 mL·min-1,内标为甲氧氯普胺,检测波长为240 nm.结果:吗氯贝胺在40～4 000 μg·L-1范围内线性关系良好(r=0.999 9,n=8),血浆中最小检测浓度为10 μg·L-1(S/N=3),提取回收率为96.48%～100.38%,方法回收率在99.39%～103.07%之间,日内RSD≤8.14%,日间RSD≤6.25%.结论:该方法快速、准确、灵敏度高、专一性强,可用于吗氯贝胺血浆浓度监测和药动学及生物利用度研究.     

LC-MS/MS法测定伊曲康唑的血浆浓度及在生物等效性试验中的应用(英文)

目的建立液相色谱-串联质谱(LC MS/MS)法测定人血浆中伊曲康唑浓度方法,并评价伊曲康唑受试制剂和参比制剂是否具有生物等效性。方法采用蛋白沉淀的前处理方法,以10μL进样分析。色谱柱为CAPCELLPAKC18柱,流动相为V(乙腈)∶V(水)∶V(甲酸)=75∶25∶0.15),流速为0.20 mL.min-1。串联质谱API 3000采用正电喷雾离子源和多反应监测的扫描模式。结果伊曲康唑在质量浓度2～500μg.L-1内线性良好,定量下限为2μg.L-1,批内、批间的RSD均<10%。伊曲康唑受试制剂和参比制剂的相对生物利用度为(112.71±59.58)%,经方差分析及双单侧t检验表明两者具有生物等效性。结论该方法可以准确、快速和灵敏地应用于伊曲康唑的药动学与生物等效性研究,结果显示受试制剂和参比制剂生物等效。     

反相高效液相色谱法测定氯普噻吨片的含量

目的建立一种反相高效液相色谱法测定氯普噻吨片含量的方法。方法色谱条件为:以Shim-Pack C18(150 mm×4.6mm,5μm)柱为分析柱,甲醇-0.1 mol.L-1盐酸液(60∶40)含0.008 mol.L-1十二烷基硫酸钠为流动相,紫外检测波长328nm,流速为1.0 mL.min-1。结果氯普噻吨浓度在10~90μg.mL-1范围内峰面积与浓度线性关系良好(r=0.999 9);平均加样回收率为99.5%,RSD为0.48%;日内RSD为0.40%,日间RSD为0.56%(n=5)。结论本方法准确、简便、可靠,适用于测定氯普噻吨片的含量。     

氢溴酸加兰他敏口腔崩解片的人体药代动力学和生物等效性研究

目的建立人血浆中加兰他敏的高效液相色谱-质谱测定方法,用于研究氢溴酸加兰他敏口腔崩解片的人体药代动力学和生物等效性。方法20名男性健康志愿者随机交叉给药,分别单剂量口服国产的氢溴酸加兰他敏口腔崩解片(受试制剂)及氢溴酸加兰他敏口腔分散片(参比制剂),采用高效液相色谱-质谱法,电喷雾电离源选择性正离子检测受试者血浆中加兰他敏的浓度。结果计算两者主要药动学参数:Cmax分别为(51.883±14.185)和(53.500±13.478)μg.L-1,tmax分别为(1.092±1.014)和(1.150±0.528)h,AUC(0→30)分别为(501.679±136.094)和(464.010±100.890)μg.h.L-1。结论受试制剂对参比制剂的相对生物利用度为106.2%,两制剂在人体内具有生物等效性。     

血浆中西沙必利胶囊的HPLC法分析及生物等效性研究

目的 :建立人血浆西沙必利的反相高效液相色谱荧光检测分析方法 ,并对其在人体内的生物等效性进行研究。方法 :血浆样品在碱性条件下经三氯甲烷 -异丙醇 (9∶1)提取 ,用KromasilC18柱 ,乙腈 - 0 0 5mol·L-1磷酸盐缓冲液 -三乙胺 (39∶6 0∶1)为流动相 ,流速 1 0mL·min-1,荧光检测激发波长 2 72nm ,发射波长 35 0mm等条件下 ,分析测定。结果 :西沙必利在 2 5～ 12 0 μg·L-1浓度范围内呈良好线性关系 (r =0 9992 )。血浆最低检出浓度为 1ng·mL-1。结论 :本法准确灵敏 ,重现性好 ,可用于西沙必利的药代动力学研究。     

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.