反相高效液相色谱法测定大鼠血浆及子宫组织中达那唑的药物浓度

目的:建立反相高效液相色谱法测定大鼠血浆及子宫组织样品中达那唑药物浓度。方法:生物样本经沉淀蛋白后,上清液直接进样。色谱条件:甲醇-水(78:22)为流动相,用 Agilent Zorbax Eclipse XDB-C_(18)(250 mm×4.6 mm,5 μm)色谱柱进行分离,284 nm 检测,流速1.0 mL·min~(-1)。结果:血浆中达那唑的线性范围为31.25～2500 ng·mL~(-1),定量限为31.25 ng·mL~(-1);子宫组织中达那唑的线性范围为62.5～2500 ng·g~(-1) ,定量限为62.5 ng·g~(-1);方法回收率均人于90%。结论:本方法简单、准确,专属性强,灵敏度高,可用于临床药代动力学研究。     

吲哒帕胺血药浓度测定及药动学研究

目的:建立一种测定人体血浆中吲哒帕胺血药浓度的高效液相色谱法。方法:采用 ULTRON VX-ODS 色谱柱(5μm,4.6 mm×250 mm),以甲醇-乙腈-水(50∶5∶45)为流动相,流速1 mL·min~(-1),柱温:室温,检测波长242 nm。结果:本方法线性范围为0.54～172.8ng·mL~(-1),r=0.9940,方法定量限为(0.54±0.04)ng·mL~(-1)(S/N>10),方法回收率为97.8%～108.1%(n=15),日内 RSD 为3.0%～13.4%(n:5),日间 RSD 为2.5%～14.4%(n=5).结论:本方法灵敏度高,操作简便准确,可用于人体血浆中吲哒帕胺浓度的测定及药动学研究.     

固相萃取HPLC测定人血浆中头孢西酮及在药动学研究中的应用

目的:建立反相高效液相色谱办法测定人血浆中头孢西酮浓度,并用于注射用头孢西酮钠人体药代动力学研究。方法:采用高效液相色谱紫外检测法,血浆中加入内标后经固相萃取,色谱柱为 Apollo C_(18)(5μm,250 mm×4.6 mm)。流动相为乙腈-0.02 mol·L~(-1)醋酸铵溶液(18∶82)(pH 5.0),流速1 mL·min~(-1),检测波长为278 nm。结果:本方法线性检测范围为0.5～250μg·mL~(-1),线性关系良好(r=0.9996);最低检测浓度为0.5μg·mL~(-1);方法绝对回收率为67.2%～84.0%,相对回收率为91.1%～102.1%;日内、日间 RSD 均小于8%。结论:本方法灵敏度高,操作简便,可用于人血浆中头孢西酮的浓度测定及临床药代动力学研究。     

液相微萃取光化学荧光高效液相色谱法测定生物样品中枸橼酸氯米芬顺反异构体的含量

目的:建立液相微萃取光化学荧光高效液相色谱法测定生物样品中枸橼酸氯米芬顺反异构体的含量。方法:应用自制的液相微萃取装置,对生物样品中的枸橼酸氯米芬顺反异构体萃取后,分离采用 Sinochrom ODS—BP C_(18)色谱柱(200 mm×4.6mm,5μm);流动相为甲醇-水(70∶30,每1L 水含0.25 mL 磷酸、50μL三乙胺);流速为0.8 mL·min~(-1);荧光检测器的激发波长和发射波长分别为255 nm 和378 nm;柱温20℃。结果:利用该方法可以将血浆、尿液和肝脏组织匀浆中的枸橼酸氯米芬顺反异构体同时提取分离,浓缩倍数可达5～7倍;枸橼酸氯米芬在血浆、尿液和肝脏中的线性范围分别为0.05～20μg·mL~(-1),0.02～20μg·mL~(-1),0.04～40μg·g~(-1);检出限分别为20 ng·mL~(-1),10μg·mL~(-1),20 ng·g~(-1);精密度试验的 RSD 小于11.7%。结论:本文将液相微萃取技术应用于生物样品中枸橼酸氯米芬顺反异构体的提取分离,利用高效液相色谱法荧光检测,获得了较好的效果。     

LC-MS/MS法同时测定人血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的质量浓度

目的建立一种简单、快速的能同时测定血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明质量浓度的LC-MS/MS方法。方法血浆经乙腈沉淀后,上清液直接进样分析。色谱柱为Hypersil GOLD aQ,流动相为乙腈-1 mL·L~(-1)甲酸溶液,梯度洗脱。正离子检测,扫描方式为选择性反应监测,氟西汀、去甲氟西汀、帕罗西汀、氟伏沙明和内标罗红霉素的检测离子对分别为m/z 310.2→44.4,296.1→134.1,330.2→192.1,319.2→259.1和837.7→679.3。结果血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的质量浓度在1～1 000 ng·mL~(-1)范围内线性关系良好,最低定量下限为1 ng·mL~(-1)。氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的日内和日间精密度RSD值均小于15%,日内和日间相对误差在-15%～15%之间,药物的提取回收率均大于90%,无显著基质效应。药物在反复冻融3次、室温下保存12 h和-80℃保存1个月的条件下稳定。该方法成功应用于抑郁症患者的氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明血药质量浓度监测。结论建立的LC-MS/MS分析方法简便、灵敏、准确,可用于临床血药质量浓度的监测。     

离子对-反相高效液相色谱法测定人胎盘和羊水中利凡诺

目的:运用离子对-反相高效液相色谱法测定人胎盘和羊水中利凡诺的含量。方法:采用 Spherisorb C_(18)色谱柱(250mm×4.6 mm,5μm);柱温:30℃;流动相:甲醇-乙腈-0.05%十二烷基磺酸钠溶液(35:35:30,pH=3);流速:1.0 mL·min~(-1);荧光检测器:激发波长360 nm,发射波长505 nm。结果:人胎盘、羊水样品分别在10 ng·g~(-1)～1μg·g~(-1)、10 ng·mL~(-1)～1μg·mL~(-1)浓度范围内,均呈现良好线性,检测限分别为3 ng·g~(-1)、3 ng·mL~(-1),准确度和精密度好。结论:本方法操作简便,结果准确可靠。     

RP-HPLC测定注射用甲硫酸新斯的明含量的方法学研究

目的:高效液相色谱法测定注射用甲硫酸新斯的明的含量。方法:SHIMADZU C_8柱(250 mm×4.6 mm,5 μm),以0.05mol·L~(-1)磷酸二氢钾(用磷酸调 pH 至3)-乙腈(87:13),流速为1.0 mL·min~(-1),检测波长为260 nm。结果:高效液相色谱法测定的线性范围为0.32～0.72 mg·mL~(-1),相关系数为0.9997;供试品溶液至少8h 内稳定(RSD=0.70%);方法精密度良好(RSD=0.60%);回收率为99.1%～99.8%,RSD 为0.54%～0.66%;重复性 RSD 为0.49%。结论:本方法结果准确,重复性好,可用于该制剂的质量控制。     

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。     

高效液相色谱法测定瑞格列奈片的含量

目的:建立高效液相色谱法测定瑞格列奈片的含量。方法:采用 Alltima C_(18)柱(150 mm×4.6 mm,5 μm),以甲醇-磷酸盐缓冲液(70:30)为流动相,流速为1.0 mL·min~(-1),检测波长为243 nm。结果:线性范围为2.104～210.4 μg·mL~(-1),r=0.9999,平均回收率(n=9)为99.7%,最低检测限为3.4 ng,最低定量限为10.5 ng。结论:该方法准确、快速、简便,适用于瑞格列奈片的含量测定。     

HPLC法同时测定人体血浆中氟尿嘧啶及其前体药替加氟的浓度

目的:建立同时测定氟尿嘧啶及其前体药替加氟在人体血浆中浓度的 HPLC 方法。方法:以阿昔洛韦为内标,血浆样品用硝酸银(10%)沉淀蛋白质,采用 Zorbax-ODS C_(18)分析柱(4.6 mm×250 mm,5μm),检测波长为265 nm,流动相:0.01 mol·L~(-1)磷酸盐(用2 mol·L~(-1)氢氧化钠溶液调节 pH 为8.0)-甲醇(97:3),流速1.0 mL·min~(-1),柱温为45℃。结果:氟尿嘧啶、替加氟分别在0.1～20μg·mL~(-1)(r=0.9998)和0.2～40μg·mL~(-1)(r=0.9998)浓度范围内线性关系良好,最低检测浓度分别为5和10 ng·mL~(-1),方法回收率分别为97.2%～101.3%和98.3%～102.6%,日内、日间 RSD 小于6%。结论:方法灵敏、快速、准确,并适用于临床上氟尿嘧啶及前体药替加氟浓度监测。     

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.