HPLC法测定人血浆中伏立康唑的浓度

目的:建立一种快速、灵敏、准确、稳定的测定人血浆中伏立康唑浓度的方法,用于伏立康唑临床治疗药物监测。方法:采用高效液相色谱(HPLC)-紫外检测法,以卡马西平为内标,乙腈蛋白沉淀法处理血浆样品。色谱柱为Phecda C18,流动相为20mmol/L磷酸二氢钾缓冲液(p H=6.0)-乙腈(50∶50,V/V),流速为1 ml/min,柱温为40℃,检测波长为255 nm,进样量为50μl。结果:伏立康唑和卡马西平的保留时间分别为8.34和6.24 min;血浆中伏立康唑线性范围为0.10~20.00μg/ml(r=0.999 5),定量下限为0.05μg/ml,日内、日间RSD分别为≤1.57%、1.45%,低、中、高梯度浓度提取回收率在81.40%~128.29%之间。结论:该方法操作简便,结果准确,适用于伏立康唑的临床治疗药物监测。     

高效液相色谱法测定血浆中伏立康唑的浓度

目的建立测定人血浆中伏立康唑浓度的高效液相色谱法。方法色谱柱为Hedera ODS-2(4.6 mm×250mm,5μm),流动相为:0.1 mol·L-1磷酸二氢钠-水-乙腈(20∶35∶45),流速为1.0 mL·min-1,血浆样品经乙酸乙酯-二氯甲烷(75∶25)提取后,在255 nm波长下进行检测。结果伏立康唑在0.101~12.16μg·mL-1与峰面积呈现良好的线性关系(Y=0.5345X+0.00254,r2=0.9995),相对回收率均在95%~110%,日内和日间RSD均<6%。结论该方法专属性强、灵敏度高,适合伏立康唑的血浆浓度测定及药动学研究。     

高效液相色谱法测定人血清中伏立康唑的质量浓度

目的建立测定人血清中伏立康唑浓度的高效液相色谱法,用于临床该药的治疗药物浓度监测。方法采用乙腈沉淀处理血清样品,采用高效液相色谱紫外检测法测定。以硝西泮为内标,色谱柱为Hypersil ODS柱（200 mm×4.6 mm,5μm）,流动相为25 mmol/L磷酸二氢钾缓冲液-乙腈（32∶68）,流速为1.0 m L/min,柱温为35℃,波长为256 nm。结果伏立康唑质量浓度在0.3-12μg/m L范围内与峰面积比线性关系良好（r=0.999 3）,低、中、高质量浓度回收率分别为（105.0±3.6）%,（101.9±3.3）%,（101.1±3.4）%;日内、日间精密度的RSD均小于5%,定量限为0.3μg/m L。结论该方法简便快速、准确,适用于伏立康唑血药浓度的临床监测。     

高效液相色谱法测定人血浆中伏立康唑的含量

目的建立高效液相色谱法测定人血浆中伏立康唑浓度的方法学。方法色谱柱:ZORBAX Eclipse Plus C₁₈,柱温:40℃,流速:1 mL·min^-1,流动相:甲醇-水=55∶45,检测波长:254 nm,内标:对氯苯乙酰胺。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、稳定性。结果血浆中内源性杂质对样品测定无干扰,血浆中伏立康唑在0.1~10.0μg·mL^-1内线性良好(r=0.999 2),最低检测浓度为0.1μg·mL^-1。血浆中伏立康唑日内、日间RSD均<10%,提取回收率为89%~92%。结论本方法简便、灵敏、准确、高效,适用于人体内伏立康唑的浓度监测。     

RP-HPLC法测定肾移植术后患者血浆中伏立康唑的浓度

目的:建立一种快速检测肾移植术后患者血浆中伏立康唑浓度的方法。方法 :血浆样品处理后,采用反相高效液相色谱(RP-HPLC)检测。色谱柱为Agilent HC-C18,流动相为0.05 mol/L磷酸二氢钠(pH 6)-乙腈(40∶60),流速为1.0 ml/min,柱温为40℃,进样量为20μl,测定波长为254 nm。结果:伏立康唑血药浓度在0.258²5.8 mg/L范围内线性关系良好(r=0.999 8),定量下限为0.258 mg/L;最低检测限为0.085 mg/L;3种浓度样的日内、日间RSD<6%;平均回收率在95%25.8 mg/L范围内线性关系良好(r=0.999 8),定量下限为0.258 mg/L;最低检测限为0.085 mg/L;3种浓度样的日内、日间RSD<6%;平均回收率在95%¹20.0%之间。并采用此方法测定3例肾移植患者服用伏立康唑10 d血浆中的伏立康唑的浓度分别为4.826、4.045、1.564 mg/L。结论:本方法操作简便、结果准确,适用于人血浆中的伏立康唑药物浓度的检测。     

高效液相色谱法测定人血浆中伏立康唑的浓度

目的:建立检测血浆中伏立康唑浓度的高效液相色谱(HPLC 法。方法:采用Kromasil 100-5 C18色谱柱(250 mm×4.6 mm,5 μm),流动相为乙腈-0.02 mol/L四丁基硫酸氢铵溶液(50∶50),流速1.0 mL/min,柱温30℃。结果:伏立康唑在0.1~8.0 μg/mL血药浓度范围内与峰面积线性关系良好,回归方程为R=1.077 9C-0.040 7,r2=0.999 5。日内和日间精密度的RSD均不超过10%,定量限为0.066 μg/mL。结论:该方法完全满足临床实验的需要和相关指导原则的要求,可为临床伏立康唑血浆浓度检测和药动学研究提供参考。     

高效液相色谱法测定人血浆中伏立康唑浓度

目的建立一种快速、灵敏及检测范围广的高效液相色谱法测定人血浆中伏立康唑浓度的方法.方法采用LC-10A(岛津)高效液相色谱仪,以尼群地平为内标,以Hypersil ODS柱(250mm×4.6 mm,5μm)为分析柱;流动相为20 mmol·L-1磷酸二氢钾缓冲液(pH 6.0)-乙腈(5545);流速为1.0 mL·min-1;进样量为50μL;检测波长为254 nm.血浆样品用乙腈沉淀蛋白,离心后取上清液进样分析.结果色谱峰分离良好,无内源性杂质干扰.线性方程为Y=0.312 4X-0.069 3,r=0.999 1(0.2～20.0 mg·L-1);最低检测浓度为0.05 mg·L-1.平均方法回收率为94.6%～105.4%(n=15),日内、日间RSD≤9.2%.结论本方法准确灵敏,操作简便、快速,适用于伏立康唑血药浓度的监测,也可用药动学研究.     

LC-MS/MS法测定兔血浆中伏立康唑的浓度

目的建立LC-MS/MS法测定兔血浆中伏立康唑的浓度,并应用伏立康唑滴眼液在兔体内的药动学研究。方法色谱柱为CAPCELL CORE ADME C18柱(100 mm×2.1 mm,2.7μm),流动相为乙腈-体积分数为0.1%的甲酸水溶液(体积比为60:40),流速为0.2 m L·min-1,采用乙醚-二氯甲烷(体积比为3:2)萃取法处理血浆样品,以多反应监测模式(multiple reaction monitoring,MRM)检测,测定兔单眼单次给药后血浆中的伏立康唑浓度。结果伏立康唑质量浓度在0.5~100.0μg·L~(-1)内线性关系良好(r=0.995 4),定量下限为0.5μg·L~(-1),日内和日间精密度均小于8.9%,准确度在(-8.0~1.1)%内,平均提取回收率(96.8±7.0)%,基质效应为(98.1±3.9)%。伏立康唑滴眼液在家兔血浆中的主要药动学参数:t_(1/2)为(2.9±0.7)h,ρmax为(50.8±12.7)μg·L~(-1),tmax为(0.13±0.10)h,AUC0-12h和AUC0-∞分别为(57.9±21.4)和(60.7±21.8)μg·h·L~(-1)。结论该方法适用于伏立康唑滴眼液在兔体内的药动学研究。     

HPLC法测定人血清伏立康唑浓度

目的 建立高效液相色谱(high performance liquid chromatography,HPLC)测定人血清中伏立康唑的方法,为临床伏立康唑的个体化用药提供方法学基础.方法 采用7%高氯酸直接沉淀血清蛋白,色谱柱采用汉邦Hedera ODS-2(4.6μm×250 mm,5μm),流动相为乙腈-20 mmol/L磷酸二氢钠缓冲液(pH=6.0)(60:40,v/v),流速为1.0 mL/min,检测波长为254 nm.结果 伏立康唑在0.367~23.5μg/mL间线性良好(r=0.9964),批内、批间准确度偏差<±5.0%,批内、批间精密度相对标准偏差(relative standard deviation,RSD)<11.1%,在室温放置、冰冻、反复冻融条件下考察样品稳定性,伏立康唑均保持稳定,偏差<±12.5%.结论 建立的HPLC测定人血清伏立康唑浓度的方法具有操作简便、特异性高、灵敏度高、成本低的特点,更适合临床中常规应用.     

RP-HPLC法测定AIDS合并肺部真菌感染患者伏立康唑的血药浓度

目的建立测定人血浆中伏立康唑浓度的RP-HPLC法,并测定AIDS合并肺部真菌感染患者伏立康唑的血药浓度。方法留取患者稳态谷和稳态峰浓度时的血样,以岛津LC-20A高效液相色谱仪进行测定。色谱柱为DiamonsilC18柱,流动相为0．01mol·L-1醋酸铵-乙N（50：50,V／V）,流速0．8mL·min-1,紫外检测波长255nm,内标为酮康唑,柱温40℃,进样量50uL。结果色谱峰分离良好,血浆内源性杂质无干扰,回归方程为Y=8．4002x＋2．26×10-2（r=0．9999）,血浆中伏立康唑在0．10～19．87mg·L-1内线性关系良好,日内和日间精密度（RSD）均〈5％。结论本方法灵敏、准确、重现性好,适于伏立康唑血药浓度的测定。     

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.     

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.