HPLC测定非冷冻人全血中吲达帕胺及相对生物利用度

目的：建立人全血中吲达帕胺的HPLC—UV测定法，研究2种吲达帕胺片的相对生物利用度。方法：全血样品液液萃取后，经Diamonsil C18柱（4．6mm×200mm，5μm）分离，240nm检测，流动相为乙腈-0．1％三乙胺溶液（磷酸调pH至4．0）（38：62，v／v），流速为1．0mL·min^-1。18名健康志愿者采用随机交叉方式分别单次口服吲达帕胺片T或R 5mg，在不同时间点取血，样品以新建立的HPLC—UV法测定，研究比较2制剂的药动学及相对生物利用度。结果：吲达帕胺与全血中内源性杂质分离度好，吲达帕胺浓度在3．1～500μg·L^-1范围内与峰面积比线性良好，最低定量浓度为3．1μg·L^-1。方法回收率为95．6％-102．6％，日内精密度（RSD）小于14．2％，日间精密度（RSD）小于9．9％。2种制剂的Cmax为（250．9±84．1）μg·L^-1和（245．4±78．8）μg·L^-1；Tmax为（2．4±0．5）h和（2．4±0．5）h；AUC0-72为（4417．4±976．3）μg·h·L^-1和（4372．7±942．0）μg·h·L^-1；T1/2为（21．9±8．6）h和（21．4±5．5）h。结论：该方法选择性好，灵敏度高，可用于吲达帕胺的体内过程研究。药动学参数经方差分析表明吲达帕胺片T和R中吲达帕胺的主要药动学参数之间均无明显差异，双单侧t检验结果表明2种制剂为生物等效制剂。     

吲达帕胺缓释胶囊人体药动学和生物等效性试验

目的：研究吲达帕胺试验制剂（缓释胶囊）和参比制剂（片剂）的人体药动学和生物等效性。方法：20名健康志愿者按随机双周期交叉试验方案设计，分别口服受试制剂（吲达帕胺缓释胶囊）和参比制剂（吲达帕胺片）1．5mg，采用液相色谱串联质谱（LC／MS／MS）分析方法测定吲达帕胺的全血浓度。利用DAS程序计算其药动学参数和评价生物等效性。结果：吲达帕胺试验胶囊和参比片剂的主要药动学参数：Cmax分别为（38．2±14．6），（41．1±7．6）ug·L^-1；Tmax分别为（11．4±3．3），（8．0±3．0）h；t1/2kc分别为（19．7±2．3），（19．7±2．8）h；AUC0-96分别为（1252．8±404．3），（1267．5±278．8）ug·h·L^-1；AUC0-∞分别为（1308．0±424．6），（1319．4±292．4）ug·h·L^-1；受试制剂相对生物利用度为（97．5±14．5）％。结论：经方差分析及双单侧t检验结果显示，受试的吲达帕胺胶囊与参比的吲达帕胺片具有生物等效性。     

硝呋太尔血药浓度测定及健康志愿者药动学

目的：建立测定硝呋太尔血药浓度的高效液相色谱（HPLC）法,研究健康志愿者硝呋太尔片的人体药动学.方法：健康男性志愿者20名,单剂量口服硝呋太尔片400mg,采用HPLC法测定其血药浓度,计算其主要药动学参数.结果：硝呋太尔与内标奥硝唑保留时间分别为5.1 min和6.7 min,两者峰形良好,分离完全.高、中、低浓度日内、日间变异系数均小于10%.硝呋太尔单剂口服主要药动学参数t1/2为（3.6±0.8）h,tmax为（2.33±0.29）h,Cmax为（46.4±20.2）μg·L^-1,AUC0-12为（143.4±36.3）μg·L^-1·h^-1,AUC0-∞为（159.2±44.4）μg·L^-1·h^-1.结论：硝呋太尔片剂主要药动学参数与国外文献报道相近,提示地域性和种族性差异无显著性;本试验建立的高效液相硝呋太尔测定方法灵敏、可靠,适用于其血药浓度检测和人体药动学研究.     

盐酸罗匹尼罗片的人体药动学

目的：研究盐酸罗匹尼罗片剂在健康人体的药动学。方法：以液相色谱-串联质谱分析法（LC-MS-MS）测定12名健康志愿受试者单次口服1．0mg罗匹尼罗片后36h内不同时间的血药浓度，采用DAS2．0．1软件计算给药后的药动学参数。结果：12名健康受试者单次空腹口服1．0mg罗匹尼罗片后，其主要药动学参数t1／2为（6．7±1．1）h；tmax为（1．4±0．5）h；Cmax为（1．6±0．5）μg·L^-1；AUC0-t为（10．8±3．1）μg·h·L^-1，AUC0-∞为（11．1±3．2）μg·h·L^-1，MRT为（8．5±1．1）h，CL／F为（95．8±21．9）L·h^-1，Vd／F为（916±256）L。结论：本方法灵敏、快速、准确、选择性强，可较好地满足罗匹尼罗的药动学研究，为临床用药提供依据。     

奥美沙坦酯胶囊的人体药动学及生物等效性

目的：研究奥美沙坦酯胶囊与片剂的人体药动学及相对生物利用度.方法：18名健康志愿受试者采用随机交叉试验设计,分别口服受试制剂（奥美沙坦酯胶囊）和参比制剂（奥美沙坦酯片）20 mg,服药后0.5～36 h内间隔采血,采用固相萃取结合HPLC-MS/MS法,比较两者的生物利用度.结果：奥美沙坦酯胶囊与片剂的主要药动学参数：Cmax分别为（530±180）μg·L^-1和（660±240）μg·L^-1;tmax分别为（3.1±0.8）h和（2.4±0.7）h;AUC0-36h分别为（4 200±1 700）h·μg·L^-1和（4 400±1 400）h·μg·L^-1;t1/2分别为（6.7±1.1）h和（6.7±0.9）h;以AUC0-36 h计算,奥美沙坦酯胶囊的相对生物利用度为（95.6±22.4）%.结论：奥美沙坦酯胶囊与片剂具有生物等效性.     

LC-MS/MS法测定克拉霉素血浓度及药动学

目的建立测定人血浆中克拉霉素的LC-MS／MS法，研究市售克拉霉素胶囊在人体的药动学特点。方法以罗红霉素为内标，取血浆样品0．3mL经蛋白沉淀后，以甲醇-水（1％甲酸溶液）-乙腈=（80：10：10）为流动相，用C18柱分离，采用电喷雾离子源，正离子方式检测，扫描方式为多反应检测（MRM）。结果克拉霉素线性范围为10～2500μg·L^-1，日内、日间RSD均〈3．5％。应用此法研究18名健康受试者单剂量口服250mg克拉霉素市售胶囊的药动学参数tmax、ρmax、t1/2、AUC0→t和AUC0→∞，其值分别为（1．95±0．71）h、（949±399）μg·L^-1、（4．79±0．84）h、（5600±1753）μg·h·L^-1、（5766±1776）μg·h·L^-1。结论该法用于人体药动学的研究具有专属、快速、灵敏等特点。     

地红霉素肠溶微丸胶囊的生物等效性

目的：建立准确、灵敏的HPLC-MS测定人血浆中红霉素胺浓度的方法，研究地红霉素肠溶微丸胶囊健康志愿者生物等效性。方法：20名健康志愿者采用两制剂双周期自身对照交叉试验设计，单剂量口服地红霉素试验制剂和参比制剂各500mg，用液相色谱-质谱法测定血浆地红霉素代谢物红霉素胺的浓度，并计算两制剂的主要药动学参数及相对生物利用度。结果：红霉素胺的浓度与吸收峰面积比值在4．5-720μg·L^-1范围内线性关系良好，线性回归方程为C=0．014A-0．0214，r=0．9998（n=5），最低定量限为4．5μg·L^-1。低、中、高3种浓度的方法回收率为98．89％，99．78％，98．65％，平均提取回收率为70．55％，70．50％，71．52％，日内、日间RSD均小于10％。单剂量口服500mg的地红霉素试验制剂和参比制剂的主要药动学参数AUG0→96（梯形法）分别为（3025．6±690．9），（2862．8±672．1）μg·h·L^-1，AUG0-∞（梯形法）分别为（3679．0±888．1），（3500．9±813．2）μg·h·L^-1；Cmax（实测）分别为（390．4±81．2）ng·mL^-1和（396．0±63．2）μg·L^-1；tmax（实测）为（3．3±0．4），（3．35±0．29）h。受试制剂的相对生物利用度为（107．6±20．2）％。结论：该方法简单，准确度高，灵敏度好，可用于地红霉素人血药浓度测定。统计分析结果表明，两种制剂的主要药动学参数之间无明显差异；双单侧t检验结果表明，两种制剂具有生物等效性。     

液-质联用法考察单硝酸异山梨酯片的人体生物等效性

目的：建立液-质联用法（LC-MS／MS）的检测方法，评价受试与参比的单硝酸异山梨酯片在健康人体的生物等效性。方法：健康志愿者20名，随机交叉试验设计，单剂量口服受试或参比制剂，给药剂量均为20mg，应用LC-MS／MS法测定各受试者给药后不同时间点的血药浓度，计算药动学参数，应用BAPP 2．0软件进行生物等效性评价。结果：受试与参比制剂的药动学参数如下，AUC0-24分别为（3．4±0．6）mg·h·L^-1、（3．3±0．7）mg·h·L^-1，AUC0-∞分别为（3．6±0．7）mg·h·L^-1、（3．5±0．7）mg·h·L^-1；tmax分别为（1．2±0．9）h、（1．0±0．6）h；Cmax分别为（464．9±108．2）μg·L^-1、（433．6±115．3）μg·L^-1；t1／2分别为（5．4±0．7）h、（5．5±0．9）h。单硝酸异山梨酯片的相对生物利用度为（106．0±16．0）％，主要药动学参数经统计学分析无显著性差异。结论：受试与参比制剂具有生物等效性。     

盐酸伊托必利缓释片在犬体内的药动学

目的：对盐酸伊托必利缓释片和分散片在家犬体内进行药动学研究。方法：6只家犬按照单剂量双周期实验设计方案给药，建立一个测定狗血中的伊托必利的高效液相色谱法。结果：2种制剂在体内均符合单隔室一级吸收模型，缓释片和分散片Cmax分别为（548．6±54．2）μg·L^-1和（798．1±42.5）μg·L^-1。tmax分别为（6．7±1．2）h和（1．83±0．25）h，应用3P97药动学程序进行计算，t1/2（Ke）分别为（6．0±0．6）h和（5．72±0．26）h，AUC0→∞分别为（6．6±0．5）mg·L^-1·h和（7．0±0．6）mg·L^-1·h。结论：盐酸伊托必利缓释片具有显著缓释作用。     

液相色谱-串联质谱法研究人体内马来酸氨氯地平滴丸的生物等效性

目的：研究健康人口服马来酸氨氯地平滴丸后的药动学和生物等效性。方法：20个健康受试者采用随机分组自身交叉对照试验设计，口服马来酸氨氯地平滴丸10mg后应用LC／MS／MS测定血浆中氨氯地平浓度，以DAS2．0软件计算其药动学参数和评价生物等效性。结果：在选定的色谱／质谱条件下氨氯地平与内标及血浆杂质分离良好，在0．2～16．0μg·L^-1范围内线性良好。相对回收率大于95．420，日内和日间RSD小于12．6%,氨氯地平受试制剂（T）和参比制剂（R）的主要药动学参数：tmax分别为（5．7±0．8）h和（6．0±1．0）h，Cmax分别为（5．3±1．4）μg·L^-1和（5．3±1．4）μg·L^-1；t1/2分别为（43．7±4．4）h和（44．6±4．3）h；AUC0→∞分别为（188．2±49．6）μg·h·L^-1和（185．0±44．8）μg·h·L^-1；用面积法（AUC0→∞）估算的马来酸氨氯地平滴丸相对生物利用度为（102．0±13．6）%。结论：用LC／MS／MS测定血浆中氨氯地平浓度，杂质无干扰，定量限低，重复性好，准确度高。受试的马来酸氨氯地平滴丸与参比的马来酸氨氯地平片（麦利平）生物等效。     

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.