氯唑西林血药浓度HPLC测定方法的研究

目的:建立氯唑西林血药浓度的高效液相色谱分析方法。方法:以苯唑西林为内标,用乙腈为沉淀剂处理人血浆样品,分析柱为 SinoChrom ODS-BP(5μm,250mm×4.6mm),流动相为0.02mol·L~(-1)磷酸二氢钠缓冲液-乙腈(65:35,用磷酸调pH 为4.6),流速1.10mL·min~(-1),柱温30℃,在225nm 波长处检测。按内标法定量。结果:血药浓度线性范围为0.25～40.0μg·mL~(-1)(r=0.9994),最低检测限0.20μg·mL~(-1)(S/N>3),提取回收率为92.5%～100.5%(n=5),方法回收率为96.2%～100.8%(n=5),日内和日间 RSD(n=5)分别为1.78%～4.05%和1.96%～7.36%。结论:本方法简便、准确、精密度高、重现性好,适用于氯唑西林人体内药代动力学研究及人体生物等效性研究。     

固相萃取亲水作用色谱法测定人体血浆中表阿霉素

目的:建立固相萃取-亲水作用色谱法(SPE—HILIC)测定人体血浆中的表阿霉素。方法:血浆样品中加入表柔红霉素作为内标,经 Oasis HLB 固相萃取(SPE)小柱萃取后进样测定,色谱柱为 Kromasil KR100—5SIL 硅胶色谱柱(250 mm×4.6mm,5μm),乙腈-甲酸铵缓冲液(40 mmol·mL~(-1),pH 2.9)(90:10)为流动相,检测波长为254 nm。结果:血浆中表阿霉素的线性范围为0.05～2.5μg·mL~(-1)(r~2=0.9991);最低检测限为0.05μg·mL~(-1);样品的回收率高于89.4%;日内及日间精密度RSD 小于7.0%。结论:方法简便,准确可靠,流动相与质谱检测器兼容,适用于血浆表阿霉素浓度测定及药代动力学研究。     

高效液相色谱-荧光法测定人血浆中西酞普兰含量

目的:建立快速灵敏的高效液相色谱-荧光检测法测定西酞普兰的血药浓度。方法:血浆样品加入内标(右美沙芬)后用乙醚提取,乙醚层经空气流吹干后用流动相定容,以40μL 进样。色谱柱为 Kromasil KR100-5 C_(18)(250 mm×4.6 mm,5μm),流动相采用0.05 mol·L~(-1),pH=4.23的磷酸二氢钠缓冲液-乙腈(63:37),流速为0.6 mL·min~(-1),荧光激发波长为240 nm,发射波长300 nm,内标法定量。结果:本法的最低定量浓度为1.3 ng·mL~(-1),在1.3～260 ng·mL~(-1)浓度范围内线性良好,r=0.9999。日内精密度为1.1%～2.3%,日间精密度为1.7%～4.0%,萃取回收率为84.4%～88.3%,准确度为86.2%～95.6%,整个分析过程在15 min 内完成。结论:该法准确、精密度高、重现性好,适用于人血浆中西酞普兰的动力学研究和生物等效性研究。     

HPLC荧光检测法测定人血浆中的罗格列酮钠

目的 采用HPLC荧光检测法测定人血浆中的罗格列酮钠.方法 用Agilent C_(18)色谱柱(150mm×4.6 mm,5 μm),以乙腈-pH3磷酸盐缓冲液(35:65)为流动相,流速1.0 mL·min~(-1),进样量20μL,血浆样品经乙醚提取后进样,荧光检测器检测波长λ_(ex)=250 nm,λ_(em)=370 nm.结果 罗格列酮钠的线性范围为10～1000 ng·mL~(-1)(r=0.9999),最低定量限为10 ng·mL~(-1)(S/N>3),日内RSD<4%(n=5),日间RSD<16%(n=5),萃取回收率>94.81%.结论 所建方法适用于临床上罗格列酮钠片的血药浓度监测及药动学的研究.     

RP-HPLC法测定人血浆中氟比洛芬的浓度

目的:建立以反相高效液相色谱法测定人血浆中氟比洛芬浓度的方法。方法:采用外标法,以乙腈沉淀法处理样品后进样测定,色谱柱为Symmetry shield C_(18),流动相为磷酸盐缓冲液(pH7.0):乙腈=75:25,流速为1.0 mL·min~(-1),紫外检测波长为247 nm。结果:氟比洛芬血药浓度在0.05～20μg·mL~(-1)范围内线性关系良好(r=0.999 1),检测限为0.05μg·mL~(-1);低、中、高浓度的回收率在96.1%～107.1%之间,日内及日间RSD均<10%,符合方法学要求。结论:本方法简便、准确、快速,适用于人血浆中氟比洛芬浓度监测及药动学研究。     

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。     

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.     

加替沙星血药浓度HPLC测定方法研究

目的:建立加替沙星血药浓度的高效液相色谱分析方法。方法:以乳酸司帕沙星为内标,采用10％高氯酸为沉淀剂处理人血浆样品,分析柱为Phenomenex(Luna,C_(18),5μm粒径,150 mm×4.6 mm);Easyguard C_(18)保护柱。流动相为0.01mol·L~(-1)磷酸二氢钾缓冲液-乙腈 (18:5,用磷酸调pH为3.05),流速1.0 mL·min~(-1),柱温25℃,在292 nm波长处检测,按内标法定量。结果:血药浓度线性范围为0.025～8μg·mL~(-1)(r=0.999 9),最低检测浓度为0.025μg·mL~(-1)(S/N>3),萃取回收率在76.31％～87.70％(n=5),方法回收率在98.3％～102.3％(n=5),日内和日间RSD分别为3.3％～5.6％和4.8％～9.8％(n=5)。结论:本方法简便、准确,精密度高,重现性好,适用于加替沙星人体内药代动力学研究及生物等效性研究。     

液相色谱-串联质谱法测定人血浆中的龙胆苦苷浓度

目的:建立测定人血浆中龙胆苦苷浓度的液相色谱-串联质谱法。方法:血浆加入内标咖啡因后经固相萃取处理,采用 RESCEK C_8柱(150 mm×2.1 mm,5μm)分离,流动相为甲醇-10 mmol·L~(-1)醋酸铵溶液-乙腈(50:40:10),流速为0.2mL·min~(-1)。样品在三级四极杆串联质谱中经 ESI 源离子化后以多反应离子监测方式测定。结果:龙胆苦苷在3～5000 ng·mL~(-1)线性良好(r=0.9985),检测限为3 ng·mL~(-1),回收率为94.4%～104.2%,绝对回收率为92.4%～98.0%,日内、日间变异(RSD)均≤15%,色谱峰保留时间为2.25 min。结论:方法灵敏、准确、快速、特异性强,适用于中药龙胆苦苷的血药浓度测定和临床药代动力学研究。     

固相萃取高效液相色谱法测定人血浆中硫酸茚地那韦浓度

目的:建立硫酸茚地那韦血药浓度的高效液相色谱分析方法。方法:以对硝基苯胺为内标,血浆样品用固相萃取,色谱柱:Kromasil C_(18)色谱柱(150 mm×4.6mm,5μm),在线过滤器;流动相为0.1%三乙胺水溶液(用20%磷酸调节 pH 为4.3)-乙腈(69.5:30.5);检测波长为210 nm;流速1.0 mL·min~(-1),柱温为25℃。结果:硫酸茚地那韦在0.05～25.60μg·mL~(-1)的范围内有良好的线性关系(r=0.9995),最低检测浓度为0.01μg·mL~(-1)(S/N>3),该方法的相对回收率为97.4%～105.0%(n=5),绝对回收率为93.9%～105.6%(n=5);日内 RSD 为2.6%～3.7%,日间 RSD 为1.4%～4.0%。结论:本方法简便,准确,灵敏,特异性强,重现性好,可用于血浆中硫酸茚地那韦的测定及人体内药代动力学研究。     

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.     

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.     

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.