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。     

UPLC-MS-MS法测定怀山药中去甲乌药碱的含量

目的:建立超高效液相色谱-串联质谱(UPLC-MS-MS)法测定怀山药中去甲乌药碱含量的测定方法。方法:样品用0.1%高氯酸溶液提取、MCX固相萃取柱净化,5%氨水甲醇溶液洗脱,洗脱液水浴上氮吹干后用50%甲醇水溶液(含0.1%甲液)复溶。采用Waters BEH C₁₈超高效液相色谱柱(150 mm×2.1 mm,1.7μm),流动相为乙腈-0.1%的甲酸水溶液进行梯度洗脱分离,柱温30℃,流速为0.3 m L·min^-1,以非诺特罗作为内标进行定量。结果:去甲乌药碱的线性范围为1～100 ng·m L^-1(r=0.999 3),检出限为0.3μg·kg^-1,定量限为1μg·kg^-1;样品加标回收率为86.8%～91.8%,相对标准偏差为2.1%。结论:本方法净化效果好,灵敏度高,线性范围广,重复性好,能够满足怀山药中去甲乌药碱含量测定要求。     

固相萃取-高效液相色谱法测定细胞内外米托蒽醌的含量

目的:建立固相萃取-高效液相色谱法测定细胞内外米托蒽醌含量的方法,并对米托蒽醌普通剂型和脂质体剂型进行比较。方法:选择固相萃取柱(Bond-Elut,C_(18))提取样品,在 ZORBAX SB-C_(18)(4.6mm×250mm,5μm)分离柱上,以甲醇-0.02mol·L~(-1)磷酸二氢钾溶液(36:64,磷酸调 pH=3.0)为流动相,紫外检测波长242nm 处检测样品。结果:培养液米托蒽醌样品在0.1～4.0μg·mL~(-1)范围内线性关系良好,3个浓度的平均回收率为98.40%,日内 RSD 为1.12%～1.30%,日间RSD 为1.27%5～2.00%;细胞内液米托蒽醌样品在0.5～5.0μg·mL~(-1)范围内线性关系良好,3个浓度的平均回收率为91.80%,日内 RSD 为2.50%～2.70%,日间 RSD 为2.80%～3.80%。结论:本法简便、准确,适合米托蒽醌含量的测定。     

高效液相-质谱联用法测定人血浆中盐酸度洛西汀浓度

目的:建立高效液相色谱-质谱联用法测定盐酸度洛西汀在人血浆中的浓度,以研究该药物在人体内的药动学,并为临床用药提供参考.方法:以氟西汀为内标,血浆样品采用固相萃取的预处理方法,用phenomenex C18色谱柱(4.6 mm×250 mm,5μm)进行分离,以5 mmol·L-1醋酸铵(含1‰甲酸)-乙腈(55:45)为流动相,柱温40℃,流速1.0mL·min-1.采用质谱电喷雾电离源(ESI)将样品离子化,选择性正离子监测(SIM)准分子离子峰.结果:血浆中度洛西汀在0.78～93.60 μg·L-1范围内线性关系良好,r为0.999 1;最低定量质量浓度为0.78 μg·L-1;萃取回收率和方法回收率均大于92.05%;日内日间变异系数均小于15.0%.结论:本方法简便迅速,灵敏准确,选择性高,可用于度洛西汀药动学研究,并为临床用药提供部分参考依据.     

HPLC—MS／MS同时测定人血浆中4种抗HIV感染药物的浓度

目的:建立液质联用(HPLC-MS/MS)法同时测定人血浆中拉米夫定(3TC)、茚地那韦(IDV)、齐多夫定(AZT)、去羟肌苷(DDI)浓度。方法:采用 Symmetry C_(18)色谱柱(5 μm,150 mm×2.1 mm),以含有10 mmol·L~(-1)醋酸铵/0.5%甲酸水溶液和0.5%甲酸甲醇溶液为流动相进行梯度洗脱,流速0.25 mL·min~(-1),内标为替硝唑(TNZ),样品用固相萃取的方法进行处理,采用多反应监测进行定量。结果:3TC、AZT、DDI 在人血浆中0.01～10 μg·mL~(-1)范围内线性良好,IDV 在人血浆中0.02～20μg·mL~(-1)范围内线性良好;相对回收率在98.67%～103.6%之间,日内及日间精密度(RSD)均小于5%。结论:本方法能满足同时测定人血浆中3TC、IDV、AZT、DDI 浓度的测定。     

SPE—HPLC法测定菊延保康颗粒中蒙花苷的含量

目的:建立测定菊延保康颗粒中蒙花苷含量的方法。方法:选用Oasis HLB固相萃取小柱对样品进行纯化。采用C_(18)色谱柱(4.6mm×250 mm,5μm),以甲醇-0.8%磷酸(1:1)为流动相,流速为1 mL·min~(-1),检测波长为326 nm。结果:蒙花苷进样量在0.0432—2.16μg的范围内与峰面积呈良好的线性关系,r为0.9999,平均回收率为100.4%。结论:本方法操作简便快速,结果准确可靠。     

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。     

气相色谱-三重四极杆串联质谱法检测桑叶和人参中35种农药残留量

目的 采用直接处理和固相萃取前处理方法联合气相色谱-串联三重四极杆质谱法(Gas chromatography-triple quadrupole mass spectrometry,GC/MS/MS),建立了同时测定中药饮片桑叶、人参中农药残留的分析方法。方法 桑叶使用固相萃取法对其进行前处理,选用乙腈为提取剂,乙酸乙酯为定容溶液、石墨化炭黑氨基复合固相萃取小柱(以下简称GCB/NH₂柱)过柱;人参饮片采用直接处理法,选用乙腈提取,最后乙酸乙酯定容。两种样品经色谱柱分离后在多反应监测模式(Multiple reaction monitoring mode,MRM)下进行测定,基质加磷酸三苯酯(内标液)内标法定量。结果 该方法在2～40 ng·m L^-1(标准储备液4μg组),3～60 ng·m L^-1(标准储备液4μg组),5～100 ng·m L^-1(标准储备液4μg组)的线性范围内线性关系良好,相关系数(r)均大于0.99,加标回收率大部分在60%～130%之间,相对标准偏差(RSD)为0.3...     

固相萃取-高效液相色谱法测定人乳中甲氨蝶呤的含量

目的:建立胎盘植入产妇术后人乳中甲氨蝶呤含量测定的固相萃取-高效液相色谱法。方法:乳汁样品经C18固相萃取小柱提取后进样测定,色谱柱为Agilent ZOBAX SB-C18(4.6 mm×250 mm,5μm)柱,流动相为0.05%醋酸溶液-乙腈(88∶12),流速为1.0 mL.min-1,检测波长为306 nm。结果:乳汁中甲氨蝶呤在0.5～5μg.mL-1浓度范围内线性关系良好,高、中、低浓度的方法回收率分别为99.8%,100.9%,99.1%;RSD均小于10%。日内和日间精密度均小于5.8%。结论:本方法专属性强,灵敏度高,操作简便,适用于乳汁中甲氨蝶呤含量的测定。     

HPLC—MS法测定血浆中左旋黄皮酰胺的浓度

目的:建立测定血浆中左旋黄皮酰胺浓度的 LC-MS 方法。方法:血浆样品经醋酸乙酯提取处理后,以2%冰醋酸-甲醇(45∶55)为流动相,在1.0 mL·min~(-1)的流速下,用 Shimadzu ODS 色谱柱(150mm×4.6mm,5μm)分离,样品经电喷雾离子化后,通过 HP 1100 HPLC-MS 质谱仪液相色谱串联质谱法,以格列吡嗪为内标测定左旋黄皮酰胺的浓度。结果:血浆中左旋黄皮酰胺的线性范围为0.01-10μg·mL~(-1),最低定量浓度为10ng·mL~(-1),准确度在80%-90%之间,日内、日间精密度(RSD)在±10%之内。结论:该方法高效、准确,可用于左旋黄皮酰胺在体内的药代动力学研究及血浆中左旋黄皮酰胺的含量测定以控制质量。     

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.     

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.