HPLC法测定人全血中西罗莫司的浓度

目的建立全血中西罗莫司(SRL)的HPLC测定方法.方法以32-去甲氧基雷帕霉素为内标,全血样品先破坏BBC,取离心后的上清液用固相萃取处理.采用KromasilC18(150mn×4.6mm,5 μm)柱,柱温55℃,紫外检测波长278nm,流动相为乙腈-甲醇-水(452827,V/V/V),测定SRL浓度.结果全血中内源性杂质对样品测定无干扰.SRL在2.5～60μg·L-1范围内线性关系良好(r=0.999 0).日内和日间RSD均＜9.2%,全血样品多次冻融及提取后在24h内稳定性良好.结论该方法操作简便、灵敏、准确,适用于治疗药物监测及临床药学研究.     

西罗莫司血药浓度监测方法的建立及在血管瘤患儿的应用

目的：建立测定人全血中西罗莫司浓度的超高压液相色谱-串联质谱（UPLC-MS/MS）方法。方法：色谱柱为Agilent SB-C18（2.1 mm×50 mm,1.8 μm）,流动相为水（A）-乙腈（0.1%甲酸,B）,流速0.3 mL/min,梯度洗脱。依维莫司为内标,电喷雾离子源,多反应离子监测模式,正离子模式检测。西罗莫司和依维莫司离子对分别为m/z 936.5/409.5、980.6/389.3。同时采用该方法对我院血管瘤患儿西罗莫司血药浓度进行监测。结果：西罗莫司在0.625~40.000 ng/mL浓度范围内相对峰面积线性关系良好（R2=0.999 5）,日内、日间RSD均小于10%,提取回收率91.95%~118.50%,基质效应81.67%~107.84%。结论：本方法灵敏、准确、快速,能满足临床常规监测西罗莫司血药浓度的需求     

高效液相色谱法测定环孢素A的全血浓度

目的　建立的全血中环孢素A的HPLC测定法。方法　全血经多步处理后采用C18柱 ( 15 0mm× 4 6mm ,10 μm)分离 ;流动相为甲醇 水 ( 80∶2 0 ) ;流量为 1 0ml/min ;检测波长为 2 14nm ;柱温为 6 5℃ ;环孢素D为内标。结果　全血中杂质不干扰样品的测定 ,标准曲线线性关系良好 (r=0 998) ;最低可定量浓度为 5 0ng/ml;高、中、低 3种浓度的日间和日内变异均小于10 0 % ;相对回收率在 95 9～ 10 5 1% ;符合药物生物样品分析要求。     

西罗莫司血药浓度测定方法的建立及在肝移植患者中的应用

目的建立高效液相色谱法测定西罗莫司血药浓度,测定该药在肝移植患者中的药代动力学参数。方法以沉淀-萃取法处理全血样品,采用Hypersil ODS C18(250 mm×4.6 mm,5μm)柱,柱温为50℃,乙腈-甲醇-水(7.5:62.5:30)为流动相,流速1.2 mL/min,以32-去甲基雷帕霉素为内标,在276 nm波长处,检测西罗莫司血药浓度,并应用此方法测定5例肝移植患者的西罗莫司血药浓度。结果全血中西罗莫司浓度在2.0～50.0ng/mL范围内线性良好(r=0.999 7),平均相对回收率为99.1%(n=5)。5例肝移植患者的Cmax为(12.66±4.1)ng/mL,t1/2为(19.2±11.2)h,tmax为(1.8±0.3)h。结论本方法快速、简便、准确,灵敏度高,重现性好,可用于肝移植患者的西罗莫司临床药物监测。     

HPLC法测定白斑4号溶液中曲安奈德的含量

目的 :建立测定白斑 4号中曲安奈德含量的HPLC方法。 方法 :DiamonsilC18柱 (15 0mm× 4 .6mm ,5 μm) ,流动相为甲醇∶水 (6 9.5∶30 5 ) ,测定波长为 2 4 0nm ,流速 1ml/min ,内标为醋酸泼尼松龙。 结果 :在 5 .2 5～ 4 5 .5 μg/ml范围内线性关系良好 ,r=0 9998(n =6 )。平均回收率为 99.81% ,日内RSD <1% ,日间RSD <2 %。 结论 :HPLC方法测定曲安奈德选择性强、简便、准确、快速 ,可用于常规制剂分析测定。     

HPLC-MS/MS法测定全血中西罗莫司的药物浓度

目的:建立简便、快速地测定人全血中西罗莫司血药浓度的HPLC-MS/MS方法。方法:全血样品经硫酸锌及含有子囊霉素(内标)的乙腈溶液沉淀后,取上清液20μL进样分析。采用Symmetry-C18色谱柱(2.1 mm×50 mm,3.5μm),流动相梯度洗脱,流速为0.5 mL.min-1,质谱采用电喷雾离子化电离源,经多反应监测模式检测。结果:西罗莫司和子囊霉素的保留时间分别为2.24 min和2.21 min,空白血浆中内源性物质不干扰测定;西罗莫司的血药浓度在1~30 ng.mL-1时,线性关系良好(r=0.999 5);日内、日间RSD值均小于10%,提取回收率为86.4%~90.4%。结论:本方法灵敏、准确、简单、快速,可用于西罗莫司的血药浓度监测和早期肝移植患者药代动力学研究。     

HPLC法测定体液中氨溴索浓度及其药代动力学参数

建立了 HPLC测定人血浆及尿中盐酸氨溴索含量的方法 :Hypersill C18柱 (4.6mm× 2 50 mm,5μm) ,乙腈 -甲醇 - 0 .0 1 mol/L磷酸盐缓冲液 -四氢呋喃 (35∶ 35∶ 2 7.5∶ 2 .5,V/V)为流动相 ,流速 1 .5ml/min,检测波长 2 4 2 nm。结果表明 :最低检测浓度为 5ng/ml,血药浓度在 1 0～ 32 0 ng/ml范围内线性良好 ,尿药浓度在 0 .2 5～ 8.0μg/ml范围内线性良好 ,氨溴索生物半衰期为 (4.2 1± 0 .93) h。     

UPLC-MS/MS测定beagle犬内西罗莫司血药浓度

目的建立测定beagle犬内西罗莫司（SRL）血药浓度的UPLC-MS／MS联用方法。方法色谱柱为AgilentC18柱（50mm×2．1mm,1．7μm,）;预柱为C18柱（4．0mm×3．0mm,5μm）;流动相为乙腈-水（90：10,v／v）;流速为0．30ml／min;柱温为50℃;进样室温度为4℃。离子源为ESI源;正离子方式检测;扫描方式为多反应监测（MRM）;用于定量分析的离子反应分别为m／z936．6→m／z409．7（西罗莫司）和m／z826．6→m／z415．4（内标他克莫司）。取全血样品经液-液萃取后进样20μl。结果SRL在0．50～12．00ng／ml浓度范围内呈良好的线性（r=0．9997）,定量下限0．5ng／ml,日内、日间精密度（RSD）均小于15％,方法回收率均大于90％,萃取回收率大于76％。结论本方法具有样品处理简单,方法灵敏、快速、选择性强,满足西罗莫司在beagle犬体内药物检测的要求。     

高效液相色谱法测定血浆中平痛新的浓度

目的 :建立高效液相色谱法测定血浆中微量平痛新浓度的分析方法。方法 :采用HPLC法 ,色谱柱 :KromasilC18 柱 (Φ4 6mm×200mm ,5μm) ;流动相 :甲醇 -水 -三乙胺 (50∶50∶0 5) ;流速 :0 7ml/min ;检测波长 :215nm ;灵敏度 :0 04AUFS ;内标物 :奥美拉唑。结果 :血浆中平痛新浓度在10～600ng/ml范围内线性关系良好 ,相关系数r=0 9993 ;平均回收率为 (100 36±1 1) % ;平均日内和日间差异分别为4 7 %和3 8 % ;最低检测限可至1ng/ml。结论 :该法简便、稳定 ,可用于盐酸平痛新体内药代动力学与生物利用度研究。     

人血浆中吡格列酮的固相萃取高效液相色谱法测定

以吡格列酮的同系物 PIOGA为内标 ,采用 C2 固相萃取紫外检测的 HPL C分析法测定人血浆中吡格列酮的浓度。 C1 8分析柱 (15 0× 4.6 mm,5 μm) ,流动相 :水 -乙腈 -冰醋酸 (5 40∶ 46 0∶ 1.2 ,用氨试液调至 p H6 .0 ) ,流速 :1.0 ml/ min,检测波长 2 6 9nm。取血样 0 .5 m l,加入内标经 C2 固相萃取后进样 40μl。最低定量限 (L OQ)为10 ng/ ml,线性范围为 10～ 16 0 0 ng/ ml     

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.