液相色谱-串联质谱法快速测定人血浆中伊曲康唑

目的:建立灵敏、快速的液相色谱-串联质谱法测定人血浆中伊曲康唑,并用于药代动力学研究。方法:血浆样品经乙腈沉淀蛋白后,以乙腈-水-甲酸(80:20:0.2,v/v/v)为流动相,流速0.50 mL·min~(-1),Zorbax sB C_(18)柱分离,采用电喷雾电离源,以选择反应监测(SRM)方式进行正离子检测。用于定量分析的离子反应分别为 m/z 705→m/z(392 432)(伊曲康唑)和m/z 256→m/z 167(内标,苯海拉明)。结果:测定血浆中伊曲康唑的线性范围为1.00～1000 ng·mL~(-1),定量下限为1.00 ng·mL~(-1)。日内、日间精密度(RSD)均小于7.2%,准确度(RE)在±3.0%以内。应用本法测得20名健康受试者单剂量口服200mg 伊曲康唑胶囊后的主要药动学参数为:T_(max)(4.25±1.02)h,C_(max)(155.5±73.3)ng·mL~(-1),t_(1/2)(20.4±11.4)h;用梯形法计算,AUC_(0-84h)(2257±1168)ng·h·mL~(-1),AUC_(0-∞)(237.1±1207)ng·h·mL~(-1)。结论:该法选择性强、灵敏度高、操作简便,适用于伊曲康唑的临床药代动力学研究。     

犬血浆中决奈达隆及其代谢物SR35021的LC-MS/MS法测定

建立了LC-MS/MS法同时测定Beagle犬血浆中的决奈达隆及其N-去丁基代谢物SR35021,并考察了盐酸决奈达隆片在Beagle犬内的药动学。采用C_(18)色谱柱,以乙腈-甲醇-5 mmol/L乙酸铵溶液-甲酸(55:10:35:0.07)为流动相,大气压化学电离源,多反应监测(MRM)方式,正离子检测。用于定量分析的离子反应分别为m/z 557→100(决奈达隆)、m/z 501→114(代谢物SR35021)和m/z 449→252(内标,托伐普坦)。血浆中决奈达隆及其代谢物SR35021均在0.2～200 ng/ml浓度范围内线性关系良好。两者的日内和日间RSD均小于10%。主要药动学参数如下:c_(max)(495±208)和(61.6±15.9)ng/ml;t_(max)(1 67±0.61)和(1.75±0.42)h;t_(1/2)(5 92±0.59)和(3.01±0.60)h:AUC_(0-t)(2 785±1 409)和(325±137)ng·h·ml;AUC_(0-(?))(2 793±1 412)和(328±136)ng·h·m1。     

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。     

HPLC-MS法研究替米沙坦人体药动学和生物等效性

目的:建立准确、灵敏的 HPLC-MS 法测定替米沙坦在血浆中的浓度,以研究替米沙坦在健康受试者中的药动学和生物等效性。方法:20例健康受试者单次口服40mg 替米沙坦受试或参比制剂,按规定时间采集肘静脉血,乙腈沉淀处理血样。采用 Zorbax-SB-ODS 柱,乙腈-0.02mol·L~(-1)醋酸胺缓冲液(28∶72)为流动相,HPLC-MS 内标(坎地沙坦,m/z=423.3)法选择性正离子检测法测定替米沙坦(替米沙坦,m/z=515.3)血浆浓度。结果:HPLC-MS 法测定血浆中替米沙坦的最低检测限为0.5ng·mL~(-1),在2-500ng-mL~(-1)范围内线性关系良好;血药浓度测定日内、日间 RSD 均小于10%。测得替米沙坦受试和参比制剂的主要药动学参数 C_(max)(μg·L~(-1))分别为220.70±95.80和233.11±105.52,AUC_(0-96)(μg·h·L~(-1))分别为1908.54±650.43和1986.26±553.24,t_(1/2)(h)分别为24.1±4.2和24.8±3.8;T_(max)(h)分别为1.8±0.6和1.7±0.7。相对生物利用度 F 为95.6%±6.4%。结论:建立的 HPLC-MS 法灵敏、准确,统计分析表明替米沙坦受试和参比制剂生物等效。     

2种洛伐他汀片人体药动学及生物等效性研究

目的:比较2种洛伐他汀片在健康人体内的药代动力学和生物利用度,评价2种制剂的生物等效性。方法:18名男性健康志愿者随机交叉分别单剂量口服山东罗欣药业股份有限公司研制的洛伐他汀片5片(20 mg·片~(-1))或北京万生药业有限责任公司生产的洛伐他汀片5片(20 mg·片~(-1)),采用 HPLC 测定血浆中药物浓度,通过方差分析和双向单侧 t 检验比较2种制剂的 AUC_(0→24)、C_(max)、T_(max)。结果:2种制剂的 T_(max)(h)分别为2.11±0.21和2.11±0.21,C_(max)(ng·mL~(-1))为93.73±17.42和92.67±13.98,t_(1/2)(h)分别为6.75±1.33和6.50±1.09,AUC_(0→24)(ng·mL~(-1)·h~(-1))分别为480.56±55.75和478.24±69.26,AUC_(0→∞)(ng·mL~(-1)·h~(-1))分别为555.33±69.98和543.04±76.25。结论:2种洛伐他汀片生物等效,受试制剂与参比制剂的相对生物利用度为(102.67±6.98)%。     

LC／MS／MS测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度

目的:用高效液相色谱-质谱联用方法同时测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。方法:大鼠血浆用乙醚萃取法处理后,采用 LC/MS/MS 方法,测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度。HPLC 条件:大连依利特 Hypersil BDS C_(18)柱(2.1 mm×50 mm,3μm),流动相:水(含0.1%甲酸)-乙腈(39:61,v/v),柱温:12℃,流速:200 μL·min~(-1),进样量:10 μL;质谱检测参数为电喷雾电离源(ESI);喷雾电压:3500 V;碰撞压力:0.1333 Pa;源内碰撞诱导电压:普萘洛尔,10 V;4-羟普萘洛尔,15 V;N-去异丙基普萘洛尔,15 V;扫描时间:0.3 s;检测离子:普萘洛尔 m/z 260[M H]~ ;4-羟普萘洛尔 m/z 276[M H]~ ;N-去异丙基普萘洛尔218[M H]~ 。结果:普萘洛尔线性范围2—1000 ng·mL~(-1),最低定量限为2 ng·mL~(-1)。4-羟普萘洛尔和 N-去异丙基普萘洛尔的线性范围1～200 ng·mL~(-1),最低定量限为1 ng·mL~(-1)。普萘洛尔的萃取回收率在90%以上,4-羟普萘洛尔和 N-去异丙基普萘洛尔的萃取回收率均为50%以上,日内、日间 RSD 皆小于15%。结论:适用于测定大鼠血浆中普萘洛尔及其代谢物4-羟普萘洛尔、N-去异丙基普萘洛尔的浓度及药动学的研究。     

富马酸比索洛尔片在健康人体内的生物等效性评价

目的:建立人血浆中比索洛尔浓度的HPLC-MS测定方法,并评价国产与进口富马酸比索洛尔片的人体生物等效性。方法:18例男性健康受试者随机分成两组,分别交叉口服受试制剂和参比制剂各5mg,采用HPLC- MS法测定人血浆中比索洛尔的浓度。结果:血浆中比索洛尔的最低定量限为0.05 ng·mL~(-1),在0.05～120 ng·mL~(-1)范围内线性关系良好,批内及批间精密度RSD均小于8%。受试制剂与参比制剂的各主要药动学参数:T_(max)分别为(1.9±0.9)和(1.9±0.7)h,C_(max)分别为(20.3±3.3)和(20.6±3.3)ng·mL~(-1),t_(1/2)分别为(8.4±1.2)和(8.1±1.0)h,用梯形法计算AUC_(0～48h)分别为(244.0±38.5)和(249.5±41.0)ng·h·mL~(-1)。两种制剂的主要药动学参数C_(max)和AUC_(0～48h)经对数转换后进行方差分析及双单侧t检验,并计算90%置信区间,表明两种制剂生物等效,相对生物利用度为(98.4±10.6)%。结论:两种制剂生物等效。     

液-质联用法测定健康受试者血浆中他林洛尔的浓度

目的 建立液-质联用法测定人血浆中他林洛尔的浓度。方法 空白血浆加他林洛尔和内标，用乙腈直接沉淀，然后用质谱进行检测。色谱柱为Cosmosil C18 （2．0mm×150mm，5μm），柱温40℃，流动相为乙腈-10mmol·L^-1甲酸胺水溶液（含0．05％甲酸）（60：40，v／v），流速为0．2mL·min^-1，进样量为20μL,采用正离子方式扫描，他林洛尔的监测离子为m／z：364．3→308．1，内标普萘洛尔的监测离子为m/z：260．1→184．2。结果 他林洛尔的线性范围为1.00～522．40ng·mL^-1，r^2＝0．999，最小检出浓度为1．00ng·mL^-1，绝对回收率在80％左右，相对回收率在80％～115％，日内、日间RSD均〈15％。结论 本方法简便、灵敏，适用于他林洛尔血药浓度检测和药物动力学研究。     

HPLC-MS/MS法测定血浆中的他林洛尔及人体药动学研究

目的建立一种灵敏、快速、简便的测定人血浆中他林洛尔浓度的高效液相色谱-串联质谱法(HPLC-MS/MS),为研究他林洛尔在人体内的药物动力学提供手段。方法以盐酸氟西汀作为内标(I.S.),血浆样品用乙腈沉淀蛋白,离心后取上清液进样,用预柱Phenomenex C18柱(10 mm×4.6 mm,5μm),Phenomenex C18反相色谱柱(250 mm×4.6 mm,5μm),进行分离,以乙腈∶缓冲液(5 mmol.L-1醋酸胺和0.02%甲酸)(45∶55,V/V)为流动相,柱温40℃,流速:0.9 mL.min-1,HPLC-MS/MS选择m/z为364.3(母离子)和308.3(子离子)监测他林洛尔,m/z为310.0(母离子)和148.3(子离子)监测内标氟西汀。结果他林洛尔及内标盐酸氟西汀在9 min内完全分离,他林洛尔在2.0~320.0ng.mL-1时线性关系良好,相关系数为0.9976;平均萃取回收率大于77%;平均方法回收率大于91%,小于103%;最低定量限为2.0ng.mL-1;日内日间RSD均小于9%。主要药动学参数如下:Cmax(91.90±48.09)ng.mL-1;tmax(2.79±0.89)h;t21(14.67±2.15)h。结论本方法简便快速、灵敏准确,适用于他林洛尔药物动力学的研究。他林洛尔片在中国人体内的药动学参数与文献报道相似。     

LC-MS/MS方法测定大鼠血浆中亮丙瑞林浓度

目的建立迅速、简单、灵敏的液相色谱-串联质谱法测定大鼠血浆中亮丙瑞林的浓度。方法血浆样品采用固相萃取的方法,混合溶液(乙腈-水-甲酸=21.3:78.5:0.2,v/v/v)为流动相;采用Dikma C18柱(30 mm×4.6 mm,5μm)分离,通过电喷雾电离源(ESI),以选择性反应监测(SRM)方式进行正离子检测,用于定量分析的离子反应分别为m/z 605.5→221.1(亮丙瑞林)和m/z 584.5→221.1(内标,丙氨瑞林)。结果建立的人血浆内亮丙瑞林测定方法线性范围为0.02~100 ng·m L-1,定量下限可达0.02ng·m L-1。日内、日间精密度(RSD)均<13%。结论该方法预处理简洁,灵敏,专属性强,可方便用于大鼠血浆中亮丙瑞林的测定。     

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.     

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.     

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.