液相色谱-串联质谱法测定人血浆中兰索拉唑及其代谢物浓度

目的建立液相色谱-串联质谱同时测定人血浆中兰索拉唑及其代谢物兰索拉唑砜、5-羟基兰索拉唑浓度的方法。方法用Agilent SB-C18色谱柱,流动相为0.002 mol.L-1乙酸铵(用甲酸调pH为4)-乙腈(60∶40,v/v),流速为0.3 mL.min-1。用正离子电离,多离子反应监测进行定量分析。结果兰索拉唑、兰索拉唑砜、5-羟基兰索拉唑线性范围分别为5～3044,1.5～550,2～549.5 ng.mL-1,三者日内、日间精密度均小于10%,三者的提取回收率为81.55%～98.74%(RSD<10%)。结论本方法灵敏、准确、快速,可用于人血浆中兰索拉唑及其代谢物浓度的测定和药代动力学研究。     

反相高效液相色谱法检测人血浆中盐酸曲美他嗪

目的:建立人血浆中盐酸曲美他嗪检测的高效液相色谱方法。方法:血浆经乙酸乙酯-正己烷萃取,以 ZORBAX E-clipse XDB-C_(18)(4.6 mm×150 m/n,5μm)为色谱柱;流动相为乙腈-0.1 mol·L~(-1)磷酸二氢钠-0.1%三氟乙酸-水(10:10:20:60),流速为0.8 mL·min~(-1);检测波长234 nm。结果:盐酸曲美他嗪浓度在2.5～200μg·L~(-1)范围内线性关系良好(r=0.9999);最低检测限为2.5μg·L~(-1);低、中、高3个浓度的相对回收率(n=5)分别为(100.6±2.31)%,(99.84±7.53)%,(99.89±2.63)%;日内 RSD(n=5)分别为7.24%,2.61%,5.45%,日间 RSD(n=5)分别为5.64%,1.99%,2.76%。结论:本方法简便、快速、准确可靠,适用于人血浆中盐酸曲美他嗪浓度的测定及其药代动力学研究。     

反相高效液相色谱法测定人血浆中兰索拉唑的浓度

目的:建立反相高效液相色谱法测定人血浆中兰索拉唑的浓度。方法:色谱柱为Agilent Eclipse XDB C_(18)(150 mm×4．6 mm,5μm),流动相为0．01 mol·L~(-1)磷酸二氢铵(内含1‰正辛胺)-乙腈(66:34),流速为1．0 ml·min~(-1),检测波长为285 nm,内标为奥美拉唑。结果:兰索拉唑的线性范围为0．01-2．00μg·ml~(-1),回归方程为Y=0．0019C 0．0022(r=0．999 4);批内RSD为2．95％～3．25％,批间RSD为1．09％-1．95％。结论:本方法操作简便,测定结果准确,检测限较低,重现性好,可用于兰索拉唑人体药物动力学研究。     

超快速液相色谱-质谱/质谱联用法测定人血浆中兰索拉唑的浓度

目的 建立测定人血浆中兰索拉唑(抗胃及十二指肠溃疡药)含量的超快速液相色谱-质谱/质谱联用法(UFLC-MS/MS).方法 血浆样品经甲醇沉淀蛋白后,以乙腈-0.1%甲酸(27:73)洗脱,用Ultimate XB-CN(5μm,2.1mm×150mm)色谱柱,通过电喷雾离子化三重四级杆串联质谱,经多反应监测模式检测,m/z370.4→m/z252.2(兰索拉唑),m/z 237.2→m/z 194.3(卡马西平,内标).结果 兰索拉唑在5.01～5007.90μg·L-1内线性关系良好(γ=0.998 5),最低定量浓度为5.01μg·L-1.方法 回收率(n=5)分别为90.21%,89.71%,92.27%,日内和日间精密度均<9%.结论 该法操作简单,灵敏,准确,重现性好,适用于该药的临床药代动力学研究.     

液相色谱-质谱联用法测定人血浆中卢比替康的浓度

目的建立液相-质谱(LC-MS)联用方法,用于抗肿瘤药卢比替康在肿瘤患者血浆中的定量分析。方法血浆样品用0.2 mL盐酸(0.5 mol·L~(-1))酸化后,乙酸乙酯提取,上清液在40℃水浴条件下氮气流吹干,萃取物用流动相复溶后进LC-MS联用仪分析测定。采用反相C_(18)柱进行色谱分离,流动相为:乙腈:10 mmol·L~(-1)醋酸铵水溶液(90:10,V/V)。结果本方法在5～500μg·L~(-1)内线性良好,r=0.999 4(n=4),低、中、高3种浓度质控样品的批内及批间RSD均<5%,提取回收率分别为80.66%、88.47%、85.05%,最低定量限5μg·L~(-1)。结论本方法灵敏度高,操作简便易行,能够满足该药进行临床药动学研究的要求。     

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。     

HPLC法测定人血浆中的舒芬太尼浓度

目的:建立测定人血浆中舒芬太尼血药浓度的 HPLC 方法。方法:以丁丙诺啡为内标,采用正己烷-无水乙醇(19:1,v/v)进行液-液萃取。采用 Diamonsil—C_(18)柱(4.6 mm×200mm,5μm),以0.01 mol·L~(-1)KH_2PO_4-乙腈(65:35,v/v,pH 5.6)为流动相,流速为1.5 mL·min~(-1),检测波长230 nm。结果:舒芬太尼在7.8125～12500 ng·mL~(-1)范围内线性关系良好(r=0.9976),最低检测浓度为4 ng·mL~(-1)。高、中、低浓度(12500,625,31.25 ng·mL~(-1))样本方法的平均回收率均大于97%;高、中、低浓度(12500,625,31.25 ng·mL~(-1))样本日内变异分别为4.65%,6.72%,6.68%,日间变异分别为8.65%,7.49%,13.19%。结论:本方法简便,准确,稳定性好,能够满足血浆中低浓度舒芬太尼的测定及临床药代动力学研究的要求。     

LC-MS/MS法同时测定人血浆中兰索拉唑与代谢产物的浓度及其药动学研究(英文)

目的建立同时测定人血浆中兰索拉唑及其代谢产物5’-羟基兰索拉唑和兰索拉唑砜的LC-MS/MS法。方法血浆样本用乙腈沉淀蛋白后,选用Zorbax SB-C18 Narrow-Bore色谱柱(150 mm×2.1 mm,5μm),以甲醇︰10 mmol.L-1乙酸铵(65︰35,V/V)为流动相,流速为0.4 mL.min-1。选用API3200型三重四极杆串联质谱仪的多重反应监测(MRM)扫描方式进行监测,电喷雾离子化源,负离子方式。结果兰索拉唑、5’-羟基兰索拉唑、兰索拉唑砜以及内标奥美拉唑的保留时间分别为2.63、1.56、2.21、2.30 min;血浆中兰索拉唑、5’-羟基兰索拉唑、兰索拉唑砜的线性范围分别为2.00～800、1.00～400、0.200～80.0μg.L-1(r>0.99),定量下限分别为2.00、1.00、0.200μg.L-1;日内、日间相对标准差(RSD)均小于8.0%;相对偏差(RE)均在±6.0%的范围以内;提取回收率较高,且可重现;兰索拉唑、5’-羟基兰索拉唑、兰索拉唑砜在各种贮存条件下均较稳定。该方法成功地应用于兰索拉唑肠溶片在中国健康人体内的药动学研究,兰索拉唑、5’-羟基兰索拉唑、兰索拉唑砜的ρmax分别为165～1400、15.8～177、10.2～530μg.L-1,AUC0-t分别为651～7 189、99.3～639、20.5～4 372μg.h.L-1。兰索拉唑及其代谢产物的药动学存在显著的个体间差异。结论该方法快速、灵敏、专属性强、重现性好,适用于兰索拉唑及其代谢产物的人体药动学研究。     

RP-HPLC测定兰索拉唑制剂中主药及有关物质的含量

目的 建立测定注射用兰索拉唑及兰索拉唑肠溶片中主药及有关物质含量的反相高效液相色谱法.方法 以Shim-pack C18(5 μm,250 mm×4.6 mm)为色谱柱;流动相为甲醇-水-三乙胺-磷酸(620∶380∶5∶1.5);检测波长为UV 284 nm;流速为1 mL·min-1.结果 兰索拉唑在16～320 μg·mL-1(r=0.999 9)内呈良好线性关系;注射用兰索拉唑和兰索拉唑肠溶片平均回收率分别为100.0%和99.63%, RSD为分别为1.08%和1.03%(n=9).结论 该方法简便、快速、准确、重现性好,可同时适用于分离和测定注射用兰索拉唑及肠溶片中有关物质及主药含量.     

Chiral-LC-MS/MS法同时测定人血浆中兰索拉唑对映体的浓度

目的:建立Chiral-LC-MS/MS法同时测定人血浆中S-兰索拉唑和R-兰索拉唑的浓度.方法:以乙腈作为蛋白质沉淀剂,对血浆中的兰索拉唑对映异构体进行Chiral-LC-MS/MS分析.色谱柱:Chiralpak IC柱(4.6 mm×150 mm,5μm),柱温为30℃,流动相为10 mmol·L-1醋酸铵(含0.05%醋酸)-乙腈(50:50,v/v),兰索拉唑对映异构体在9.0 min内可达到基线分离.采用AB QTRAP 5500质谱仪,使用电喷雾离子化(ESI)正离子模式,兰索拉唑和内标(埃索美拉唑)的多反应监测(MRM)扫描离子对分别为m/z 370.1→252.1和346.1→198.1.结果:该方法无明显的基质效应,S-兰索拉唑和R-兰索拉唑在标准曲线范围内线性关系良好,标准曲线的线性范围均为20.00~10000 ng·mL-1,准确度在±15%,批内和批间精密度均小于10%.在兰索拉唑对映体的储存、 处理和分析过程中没有发现两者间的手性转化,测定过程中各化合物稳定性符合要求.结论:建立了简单、高选择性的Chiral-LC-MS/MS方法,可用于同时测定人血浆中的兰索拉唑对映体的浓度.     

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.