UPLC-MS/MS法测定小鼠血浆尼古丁浓度的不确定度评定

目的评定液质联用法(UPLC-MS/MS)测定小鼠血浆中尼古丁浓度的不确定度。方法分析UPLC-MS/MS法在测定小鼠血浆中尼古丁浓度时的不确定度来源,根据实验数据计算各不确定度分量并进行合成和扩展。结果小鼠血浆中尼古丁低浓度6 ng·m L~(-1)和高浓度300 ng·mL~(-1)的扩展不确定度分别为0.65 ng·mL~(-1)和20.26 ng·mL~(-1)(P=95%,k=2)。结论 UPLC-MS/MS法测定小鼠血浆中尼古丁浓度的不确定度在低浓度时主要由回收率、基质效应、重复性、生物样品配制和标准液配制引入;在高浓度时主要由生物样品配制、标准液配制、回收率和重复性引入。     

HPLC同时测定镇痛泵中的格拉司琼-舒芬太尼及格拉司琼-曲马多-芬太尼

目的 建立同时测定格拉司琼-舒芬太尼及格拉司琼-曲马多-芬太尼含量的方法.方法 采用HPLC法,分析柱为Agilent hypersil C18柱(125 mm×4.0 nm,5μm),流动相为0.02 mol·L~(-1)磷酸二氢钾(加0.6%三乙胺,磷酸调pH4.0)-乙腈,梯度洗脱,流速为1 mL·min~(-1),检测波长为220 nm.结果 格拉司琼、舒芬太尼、曲马多、芬太尼的线性范围分别为5～50μg·mL~(-1)(r=0.9999)、0.25～4.00 μg·mL~(-1)(r=0.9994)、20～70 μg·mL~(-1)(r=0.9996)、2.5～20.0μg·mL~(-1)(r=0.9999);其平均回收率分别为99.97%、98.75%、99.80%、100.53%.结论 所建方法可同时测定格拉司琼-舒芬太尼及格拉司琼-曲马多-芬太尼混合溶液的含量,且简便、准确、灵敏.     

液质联用测定人血浆中伪麻黄碱的浓度

目的:建立液质联用法测定人血浆中伪麻黄碱的浓度。方法:采集到的血浆样品以苦参碱为内标,经0.2 mL 0.02 mol·mL~(-1)碳酸钠溶液碱化后氯仿萃取进行 LC-MS 分析。色谱条件为 Kromasil C_(18)柱(150 mm×4.6 mm,5 μm),流动相为乙腈-0.1%甲酸(13:87),流速为1.0 mL·min~(-1);质谱条件为大气压电喷雾离子源(ESI 源),正离子方式检测;扫描方式为选择离子监测(SIM),用于定量分析的离子分别为 m/z 166(伪麻黄碱)和 m/z 249(苦参碱)。结果:方法的线性范围为5～300 ng·mL~(-1),定量下限为5 ng·mL~(-1),提取回收率均大于77.9%,日内、日间精密度(RSD)均小于12.7%。结论:本方法灵敏、专属,适于伪麻黄碱人体血浆药物浓度的测定。     

LC-MS/MS法测定产妇乳汁中舒芬太尼的浓度

目的:测定产妇经静脉输注泵给予舒芬太尼的镇痛期间和镇痛结束后不同时间点的乳汁药物浓度,为临床产后镇痛过程中的母乳喂养安全性研究提供测定方法与参考。方法:采用梯度洗脱,ESI离子源,使用正离子模式,多反应离子监测(MRM),液-液萃取法,加入正己烷和乙酸乙酯(1:9,V/V)提取。结果:在3. 2-400 pg·mL~(-1)的线性关系良好(R2=0. 998),定量下限为3. 2 pg·mL~(-1),提取回收率在83. 1%~(-1)00. 7%。样品测得的最低浓度(停药22 h)为4. 75 pg·mL~(-1),最高浓度(用药31 h)为196. 74 pg·mL~(-1)。持续镇痛24 h后,药物浓度在27. 19~(-1)96. 74 pg·mL~(-1)之间;停药后2-20 h,乳汁中的药物浓度在7. 88-21. 58 pg·mL~(-1)之间,停药超过20 h的药物浓度在4. 75~(-1)9. 22 pg·mL~(-1)之间。结论:建立LC-MS/MS法检测,灵敏度高、速度快、操作简单、抗干扰能力强,可准确测定乳汁中微量的舒芬太尼药物浓度。     

UPLC-MS/MS测定大鼠血浆中26-OH-人参二醇浓度的不确定度评定

目的评定超高效液相色谱-串联质谱(UPLC-MS/MS)法测定大鼠血浆中26-OH-人参二醇浓度的不确定度。方法分析UPLC-MS/MS法测定大鼠血浆中26-OH-人参二醇浓度的不确定度来源,根据各分量计算出合成不确定度并进行了扩展。结果大鼠血浆中低(13.87 ng·mL~(-1))、中(188.04ng·mL~(-1))和高(824.33 ng·mL~(-1))浓度26-OH-人参二醇的扩展不确定度分别为3.63、12.41和66.61ng·mL~(-1)(P=95%,k=2)。结论 UPLC-MS/MS法测定大鼠血浆中26-OH-人参二醇低浓度样品的不确定度主要分别由线性拟合引入,中、高浓度样品的不确定度主要由基质效应引入。该法适用于评定UPLC-MS/MS法测定血浆中26-OH-人参二醇的不确定度研究,能为复杂生物样品分析过程的不确定度评定提供一定参考。     

液-液萃取RP—HPLC法测定人血浆中佐米曲普坦的浓度

目的:建立RP-HPLC法测定人血浆中佐米曲普坦浓度。方法:以替硝唑为内标,血浆样品经液-液萃取前处理后,选用Hypersil C_(18)色谱柱(5μm,200mm×5.0mm),乙腈-磷酸盐缓冲液(0.05mol·L~(-1)磷酸二氢钠,用磷酸调pH至5.0)(15∶85)为流动相,室温下在223nm处进行测定。结果:测定佐米曲普坦血药浓度线性范围为1.92-66.14ng·mL~(-1),最低定量浓度1.92ng·mL~(-1);方法回收率为90.62%-105.7%,萃取回收率为71.70%-114.75%。结论:本法分离效果良好,灵敏度高,回收率高,日内、日间误差小,能满足于人体药代动力学及生物利用度研究。     

P91024在Beagle犬体内的药物代谢动力学

目的:建立测定 Beagle 犬血浆中 P91024浓度的 HPLC 法并进行 Beagle 犬体内的药物动力学研究。方法:取犬血浆0.5mL,加甲醇0.5 mL,涡旋1 min,加2 mol·L~(-1)碳酸钠溶液50 μL,涡旋1 min,加环己烷提取血浆样品,氮气流吹干后用甲醇溶解残渣,进行 HPLC 分析。色谱柱为 Shim-pack VP-ODS 分析柱(150 mm×4.6 mm,5μm),Shim-pack GVP-ODS 预柱(10mm×4.6 mm);流动相为甲醇-水(87∶13);流速:1 mL·min~(-1);柱温:30℃;测定波长:231 nm,外标法定量。6条 Beagle 犬灌胃给予 P91024,计算主要药动学参数。结果:在10.0～4000.0 ng·mL~(-1)范围内,P91024峰面积与浓度呈良好线性关系(r=0.9998),最低定量浓度为10 ng·mL~(-1),该法的提取回收率为90.4%～93.6%(n=5),方法回收率为98.2%～104.2%(n=5),日内精密度和日间精密度分别为4.0%～6.6%和6.1%～8.4%。Beagle 犬灌服 P91024 20 mg·mL~(-1)后,其主要药物动力学参数为 C_(max)=1493.5 ng·mL~(-1),T_(max)=3.33 h,t_(1/2β)=11.68 h,MRT=8.58 h,AUC_(0-∞)=9305.1 ng·h·mL~(-1)。结论:本法灵敏、准确,适用于 P91024的血药浓度测定及药物动力学研究。     

SPE-HPLC法测定人血浆中喹那普利和代谢产物及其人体药代动力学研究

目的:建立 SPE—HPLC 法测定人血浆中喹那普利及其代谢产物喹那普利拉的浓度,以研究喹那普利及喹那普利拉在健康志愿者中的药代动力学和相对生物利用度。方法:应用 C_(18)固相萃取柱提取血浆中喹那普利及喹那普利拉,喹那普利色谱条件为:迪马 Diamonsil C_(18)柱(150 mm×4.6 mm,5μm);流动相为乙腈-0.1%醋酸溶液(40:60,v/v);流速1.0 mL·min~(-1)。喹那普利拉色谱条件为:Phenomenex C_(18)柱(150 mm×4.6 mm,5μm);流动相为乙腈-0.005 mol·L~(-1)十二烷基磺酸钠(磷酸调 pH 2.5)(40:60,v/v);流速1.0 mL·min~(-1)。人体药代动力学试验采用单剂双周期交叉设计方案,将18名志愿者随机分为两组,分别口服参比制剂喹那普利片和试验制剂喹那普利胶囊各40 mg,清洗期为1周。结果:喹那普利的线性范围为10～800ng·mL~(-1),r=0.9931,最低定量限为10 ng·mL~(-1);提取回收率与方法回收率分别为82.1%～94.4%,92.6%～99.9%;日内、日间 RSD 均小于10%。喹那普利拉的线性范围为20～1200 ng·mL~(-1),r=0.9995,最低定量限为20 ng·mL~(-1);提取回收率与方法回收率分别为73.3%～94.0%,100.0%～105.9%;日内、日间 RSD 均小于7%。结论:本方法灵敏度高、特异性强、重复性好,测定结果可靠,统计学分析表明2种制剂的主要药代动力学参数无显著性差异,为等效制剂。     

UPLC-MS/MS法测定人血浆中布托啡诺、艾司氯胺酮和舒芬太尼的浓度

目的 建立一种测定疼痛患者血浆中布托啡诺、艾司氯胺酮和舒芬太尼浓度的超高效液相色谱-质谱联用(UPLC-MS/MS)方法。方法 以头孢他啶为内标,血浆样本用乙腈沉淀蛋白,色谱柱用Zorbax Plus-C18(2.1 mm×100.0 mm, 2.7μm),0.1%乙酸水溶液和甲醇为流动相进行梯度洗脱,流速为0.2 mL·min^-1,进样体积为1μL。用电喷雾离子源(ESI)、正离子、多反应监测(MRM)模式。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性及基质效应。结果 布托啡诺在0.10～50.39 ng·mL^-1线性关系良好,标准曲线为y=0.73x-0.02(r=0.999 3),定量下限为0.10 ng·mL^-1;艾司氯胺酮在0.11～52.96 ng·mL^-1线性关系良好,标准曲线为y=1.12x+0.02(r=0.999 4),定量下限为0.11 ng·mL^-1;舒芬太尼在0.11～52.75 ng·mL^-1线性关系良好,标准...     

LC-MS／MS测定人血浆中卡托普利的浓度及其药动学研究

目的:建立色谱-串联质谱法测定血浆中卡托普利的浓度,并利用该方法研究了2种国产卡托普利制剂在健康受试者中的药代动力学。方法:用对溴苯乙酰基溴(p-BPB)作为稳定剂及衍生化试剂,以利培酮为内标物,采用 LC-MS/MS 方法测定。以乙腈-0.025%甲酸(60:40)为流动相,色谱柱为 Alltima C_(18)(100 mm×2.1 mm,3.0μm),流速为0.2 mL·min~(-1)。采用大气压化学离子源(APCI),正离子扫描(ESI~ );用于定量分析的离子反应分别为 m/z 414.1→210.6(卡托普利衍生物)和 m/z 411.3→191.1(内标物利培酮)。结果:卡托普利线性范围为1.0～748.0 ng·mL~(-1),定量限为1.0 ng·mL~(-1),日内、日间精密度(RSD)均小于5.8%,平均提取回收率为(103.5±5.8)%。应用此方法研究了18名健康受试者单剂量口服2种卡托普利片50 mg 后的药代动力学特点,2种制剂的 T_(max)(h)分别为0.72±0.19和0.68±0.14,C_(max)(ng·mL~(-1))分别为343.4±132.3和333.6±94.6,T_(1/2)(h)分别为2.07±0.65和2.07±0.69,AUC_(0-t)(ng·h·mL~(-1))分别为442.5±95.6和424.9±78.3。结论:首次报道了用 LC-MS/MS 测定卡托普利衍生物(cap-p-BPB)的浓度。本方法专属、灵敏度高,血浆处理简便,可用于卡托普利制剂的药代动力学及相对生物利用度的研究。     

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.