高效液相色谱法测定大鼠血浆中伊立替康及其活性代谢产物SN-38的含量

目的:建立同时测定大鼠血浆中伊立替康(CPT-11)及其活性代谢产物7-乙基-10-羟基喜树碱(SN-38)浓度的高效液相色谱法。方法:以10-羟基喜树碱作为内标,先用7%高氯酸酸化血浆,再用7%高氯酸-乙腈(50∶50)沉淀蛋白。采用Hypersil C18色谱柱(4.6 mm×250 mm,5μm)进行分离;以0.05 mol·L-1的磷酸氢二钠-甲醇-三乙胺(50∶50∶0.025,磷酸调pH 3.0)为流动相;荧光检测波长:激发波长380 nm,发射波长550 nm。结果:大鼠血浆中CPT-11和SN-38线性范围分别是20～5000 ng·mL-1(r=0.9997)和2～500 ng·mL-1(r=0.9999)。两组分最低检出限分别为15 ng·mL-1和1.7 ng·mL-1。2组分平均相对回收率分别是98.7%和99.9%;平均绝对回收率分别87.2%和94.7%。2组分日内、日间精密度均小于12%。结论:本方法快速、简便、准确,灵敏度高,可用于CPT-11及其活性代谢产物SN-38药代动力学的研究。     

高效液相色谱法测定血中伊立替康及活性代谢物SN-38浓度

目的：建立高效液相色谱法同时测定结直肠癌患者血中的伊立替康（CPT-11）及其活性代谢物7-乙基-10-羟基喜树碱（SN-38）的浓度,并对我院基因型指导给药方案进行评价。方法：以2μg·mL-110-羟基喜树碱作为内标,先用100μL 10%高氯酸沉淀蛋白,再用50μL 10%高氯酸酸化血浆。采用Agilent ZORBAX Eclipse C8色谱柱（4.6 mm×150 mm,5μm）对CPT-11和SN-38进行分离;以0.05 mol·L-1的磷酸二氢钠-乙腈-三乙胺（75∶25∶0.1,v∶v,磷酸调pH 3.0）为流动相;荧光检测波长：激发波长380 nm,发射波长550 nm。结果：人血浆中CPT-11和SN-38线性范围均为3~1000 ng·mL-1,定量下限为3 ng·mL-1;准确度分别是98.5%和100.0%;回收率分别是83.8%和84.3%。结论：本方法可靠、简便、快速,可为伊立替康个体化给药提供参考。     

HPLC法测定兔血浆中羟基喜树碱浓度

建立测定兔血浆中羟基喜树碱浓度的高效液相色谱-紫外检测法.色谱柱:Lichrospher C18柱(250mm×4.6mm,5μm),C18预柱(10mm×4.6mm,5μm);流动相:乙腈-0.075mol·L-1醋酸铵缓冲液(pH6.4)(30:70),含5mmol的三乙胺;流速:1.0ml·min-1;检测波长:384nm.羟基喜树碱保留时间为5.0min,线性范围为40～1600ng·ml-1,最低检测限为25ng·ml-1.血浆中羟基喜树碱的回收率为96.32%～106.1%.本法简便实用,定量准确,可用于羟基喜树碱药代动力学研究.     

RP-HPLC法测定小鼠血浆中10-羟基喜树碱的浓度

目的:建立测定小鼠血浆中10-羟基喜树碱浓度的方法。方法:33只小鼠灌胃给予10-羟基喜树碱80mg·kg-1,于给药前及给药后0.17、0.33、0.67、1、1.5、2、4、8、12、24h时心脏取血,采用高效液相色谱法,测定其中血药浓度。色谱柱为Luna C18,流动相为水-甲醇,梯度洗脱,流速为1.2mL·min-1,内标为喜树碱,荧光检测器检测,激发波长为383nm,发射波长为553nm。结果:内源性杂质不干扰测定,10-羟基喜树碱检测浓度线性范围为1.25～80ng·mL-1(r=0.9997),平均方法回收率为96.46%～101.87%,平均萃取回收率为82.4%～109.6%,日内、日间RSD均≤5.00%。结论:该方法简便可靠、专属性好、精密度高,可用于10-羟基喜树碱的血药浓度检测。     

高效液相-荧光检测法测定兔血浆中羟基喜树碱的含量

目的建立高效液相-荧光检测法测定兔血浆中羟基喜树碱含量的方法学。方法色谱柱为D iscovery C18(15cm×4.6mm,5μm),流动相为柠檬酸缓冲液-乙腈-75nm o.lL-1-磷酸二氢钾(70∶23∶7),75nm o.lL-1磷酸二氢钾中含1%三乙胺。流速为1.0m.lm in-1,柱温为40℃,荧光检测波长为λex363nm和λem 530nm。结果该方法的线性范围为13.7656~1957.4468ng.mL-1(r=0.9993)。提取回收率和方法回收率分别为80.90%~103.59%,102.72%~108.16%.日内RSD 7.49%,日间RSD 9.40%。最低检测限为5.2ng.mL-1。结论该方法专属性强,重现性好,操作简便,适用于羟基喜树碱的药代动力学研究和血药浓度监测。     

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.     

反相高效液相色谱法测定大鼠血浆中9-硝基喜树碱浓度

目的建立一种简单、快速测定9-硝基喜树碱血药浓度的反相高效液相色谱法。方法血浆样品加喜树碱作内标,用正己烷-二氯甲烷-异丙醇(100∶50∶5)提取,氮气吹干,残渣用流动相溶解进样。色谱条件:分析柱为C18反相柱,流动相为3%甲酸溶液-乙腈(体积比65∶35),流速为2.0 mL.m in-1,在紫外检测波长370 nm处进行检测。结果9-硝基喜树碱及内标在5min内完全分离,检测限为25 ng.mL-1,在25~1600 ng.mL-1内线性关系良好,r=0.999 6,低、中、高浓度的回收率、日间及日内精密度均符合方法学要求。结论本法简便、快速、稳定、重现性好,适用于9-硝基喜树碱临床前药动学研究。     

高效液相-荧光检测法测定羟基喜树碱血浓度

目的:建立高效液相-荧光检测法测定人血浆羟基喜树碱的血浓度.方法:色谱柱为DiscoveryC18(15cm×4.6mm,5μm),流动相为枸橼酸缓冲液-乙腈-75nmol·ml-1磷酸二氢钾(70∶23∶7,含0.1%三乙胺),流速为1.0ml*min-1,柱温为50℃,荧光检测波长为λex363nm和λem530nm.结果:该方法的线性范围为19.3～1957.4ng*ml-1(r=0.9995).最低检测限为5.2ng*ml-1,平均加样回收率为91.8%.结论:该方法专属性强,重现性好,操作简便,适用于羟基喜树碱的药代动力学研究和血药浓度监测.     

HPLC测定人体血浆中盐酸格拉司琼

目的 建立RP-HPLC-荧光色谱法测定人血浆中盐酸格拉司琼的含量.方法 以20名健康男性志愿者为研究对象,采用交叉给药方案,分别单剂量口服2 mg的试验制剂和参比制剂.采取液-液萃取法处理血浆样品,色谱柱为Diamonsil C18(150 mm×4.6 mm,5 μm),流动相为甲醇-0.5%三乙胺水溶液(pH 4.0)(46∶54),激发和发射波长分别为305 nm和360 nm,流速为1.0 mL·min-1.结果 盐酸格拉司琼在0.078～20.08 ng·mL-1线性关系良好(r=0.999 4),最低检测限为1.95×10-2 ng·mL-1,日内RSD<1.9%,日间RSD<3.3%,平均提取回收率为85.7%,平均方法回收率为100.0%.结论 该方法快速、准确,适用于盐酸格拉司琼的临床药动学研究和血药浓度监测.     

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。     

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.

---

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.