消糖灵颗粒中格列苯脲的人体生物等效性

目的:建立测定人血浆中格列本脲的高效液相色谱(HPLC)法,研究中成药消糖灵颗粒中格列本脲在男性健康志愿者体内的药动学行为,评价其生物利用度和生物等效性.方法:20名健康成年男性志愿者采用随机分组自身交叉对照试验设计,单剂量口服参比制剂消糖灵胶囊(6粒,含格列苯脲4.2 mg)或试验制剂消糖灵颗粒(2袋,合格列本脲4.2 mg),用HPLC法测定血浆中药物浓度.结果:试验制剂和参比制剂的主要药动学参数tmax分别为(2.0±0.8)h和(1.8±0.6)h,Cmax分别为(100.6±26.8)μg·L-1和(114.5±28.6)μg·L-1,AUC(0-36)分别为(567.2±270.9)μg·L-1·h和(537.6±255.5)μg·L-1·h;t1/2分别为(5.3±4.8)h和(4.1±4.0)h;以AUC(0-36)计算的试验制剂的相对生物利用度为(115.7±54.9)%.结论:建立的分析方法准确灵敏,测得的数据可靠,统计学分析表明两种制剂生物等效.     

超高效液相串联质谱法测定大鼠格列美脲及其代谢物的血药浓度

目的建立超高效液相串联质谱法快速测定大鼠体内格列美脲及其代谢物羟基格列美脲的浓度。方法用乙腈沉淀蛋白的方法处理血浆,色谱柱为ACQUITY UPLC HSS T3柱(50 mm×2.1 mm,1.8μm);流动相为乙腈-0.1%甲酸水,梯度洗脱;流速为0.4 m L·min-1;用正离子多离子反应监测(MRM)扫描,内标为甲苯磺丁脲。结果血浆中格列美脲和羟基格列美脲的线性范围为10~800μg·L-1和1~80μg·L-1(r=0.999 9和0.999 5),最低定量限为2.00μg·L-1和0.50μg·L-1,回收率均为94.05%~105.33%。两者的日内、日间精密度RSD均<8.20%。结论该方法操作简便、快捷,灵敏度高,适于大鼠体内格列美脲及其代谢物的药动学研究。     

液相色谱-串联质谱法测定人血浆中格列本脲

目的:建立测定人血浆中格列本脲的液相色谱-串联质谱法,并用于临床药代动力学研究。方法:血浆样品经液-液萃取后,以乙腈-水-甲酸(90:10:0.2)为流动相,采用 Zorbax SB-C_8 柱(150mm×4.6mm,5μm)分离,通过大气压化学电离源四极杆串联质谱,以选择反应监测(SRM)方式进行检测。用于定量分析的离子反应分别为 m/z494→369(格列本脲)和m/z 324→127(内标格列齐特)。结果:LC-MS/MS 法测定人血浆中格列本脲的线性范围为0.50～500 ng·mL~(-1),定量下限为0.50 ng·mL~(-1)。以3个浓度水平的质量控制样品求得各浓度水平日内、日间精密度(RSD)均小于5.4%,相对偏差(RE)在±2.3%以内。在临床药代动力学研究中,应用此法测试了20名受试者口服盐酸二甲双胍格列本脲胶囊后血浆中格列本脲的浓度。结论:该法灵敏、快速、准确,操作简便,线性范围宽,适用于临床药代动力学研究。     

高效液相色谱法测定人血浆中芬太尼浓度

目的:建立高效液相色谱测定人血浆中芬太尼浓度的方法.方法:采用外标法,以Kromasil-C18(4.6 mm×250 mm,5μm)为固定相,舍0.02 mol·L-1磷酸二氢钠水溶液-乙腈(65:35)为流动相,流速1 mL·min-1,检测波长为220 nm.结果:血浆芬太尼浓度在6～300μg·L-1范围内与峰面积呈良好线性关系(r=0.999 6),最低检测浓度为5 μg·L-1,方法回收率为(92.6±1.5)%,提取回收率为(84.9±1.4)%,日内RSD为1.8%,日间RSD为2.6%.结论:本方法具有较高的准确度,线性范围宽,方法灵敏,专一性好,操作简便,适用于临床芬太尼血药浓度的测定及临床药动学研究的要求.     

LC-MS/MS法测定可待因在Beagle犬血浆中的质量浓度

目的建立Beagle犬血浆中可待因质量浓度的液相色谱-串联质谱(LC-MS/MS)测定方法。方法色谱柱为Agela·C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-水-甲酸(体积比为15∶85∶0.5),血浆样品经甲醇沉淀蛋白处理,以多反应监测(multiple reaction monitoring,MRM)扫描方式检测,测定Beagle犬经口给予洛芬待因缓释片后血浆中可待因的质量浓度。结果血浆中可待因质量浓度在0.2²0μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%20μg·L-1内线性关系良好,日内和日间精密度RSD≤11.5%,平均提取回收率为104.1%¹09.2%,基质效应为119.2%109.2%,基质效应为119.2%¹28.3%。可待因在Beagle犬血浆中主要药动学参数t1/2为(2.9±0.5)h,ρmax为(8.7±3.4)μg·L-1,AUC0-∞为(28.5±7.3)μg·h·L-1。结论该方法适用于可待因在Beagle犬体内药动学的研究。     

LC-MS/MS法测定人血浆中紫杉醇的浓度

目的建立一种液相色谱-串联质谱(LC-MS/MS)的方法,测定人血浆中紫杉醇的药物浓度。方法色谱柱:Diamonsil C18柱(50 mm×2.1 mm I.D,粒径5μm,北京迪马公司),流动相:乙腈-水-甲酸(体积比为55.0∶45.0∶0.1),采用沉淀蛋白法,以多反应离子监测(multiple reaction monitoring,MRM)扫描方式进行检测,测定肿瘤患者静脉滴注紫杉醇注射液后血浆中的药物浓度。结果血浆中紫杉醇的药物浓度在20.0⁵ 000.0μg·L-1以内时,其线性关系良好,相关系数r=0.995 4;日内和日间精密度RSD≤13.4%;紫杉醇的平均提取回收率为93.6%5 000.0μg·L-1以内时,其线性关系良好,相关系数r=0.995 4;日内和日间精密度RSD≤13.4%;紫杉醇的平均提取回收率为93.6%¹08.5%,紫杉醇的基质效应为102.4%108.5%,紫杉醇的基质效应为102.4%¹05.4%,紫杉醇在人血浆中主要药动学参数如下:t1/2为(5.1±3.0)h,ρmax为(3.8±0.5)mg·L-1,AUC0-36为(13.2±2.6)mg·h·L-1,AUC0-∞为(13.3±2.8)mg·h·L-1。结论该方法适用于紫杉醇在人体内的药物动力学研究。     

复方盐酸二甲双胍片的人体生物等效性

目的研究试验制剂国产复方盐酸二甲双胍片与参比制剂格列本脲片、盐酸二甲双胍片的人体生物等效性。方法健康志愿者20名,随机双交叉单剂量口服2种制剂,2次服药间隔为2 wk。分别于服药后24 h内多点抽取静脉血,用RP-HPLC测定血浆中格列本脲和盐酸二甲双胍的浓度。血药浓度经3P97程序处理,用非房室模型估算药动学参数。结果试验制剂和参比制剂血浆中格列本脲的ρmax分别为(190.91±45.01)(、175.71±27.47)μg.L-1,tmax分别为(2.60±0.87)、(2.35±0.71)h,AUC0→24分别为(1 110.85±275.12)(、1 074.77±202.76)μg.h.L-1,AUC0→∞分别为(1 187.91±275.55)(、1 168.52±168.65)μg.h.L-1;二甲双胍的ρmax分别为(3.06±0.63)、(3.06±0.55)mg.L-1,tmax分别为(1.57±0.37)(、1.65±0.37)h,AUC0→12分别为(12.05±1.92)、(12.05±1.79)mg.h.L-1,AUC0→∞分别为(12.47±1.97)(、12.51±1.80)mg.h.L-1。以格列本脲和盐酸二甲双胍计算的人体相对生物利用度分别为(103.8±17.9)%和(100.7±13.0)%。结论2种制剂具有生物等效性。     

HPLC-MS/MS法测定人血浆中美洛昔康的浓度及药物动力学

目的建立HPLC-MS/MS法测定人血浆中美洛昔康(meloxicam)的浓度。方法色谱柱为Phenomenex Luna C18(150 mm×4.6 mm,5μm),流动相为乙腈-20 mmol·L-1醋酸铵水溶液(水中含体积分数为0.2%的甲酸)(体积比为70∶30),提取剂为乙酸乙酯-异丙醇(体积比为9∶1),内标物为吡罗昔康,离子源为ESI,流速为0.4 m L·min-1,柱温为30℃。结果美洛昔康线性为0.1⁴0.0μg·L-1(r=0.993 1),定量下限为0.1μg·L-1,日间和日内精密度RSD均小于11.7%,提取回收率在83.4%40.0μg·L-1(r=0.993 1),定量下限为0.1μg·L-1,日间和日内精密度RSD均小于11.7%,提取回收率在83.4%⁸7.1%之间。20名健康受试者背部涂抹美洛昔康凝胶剂后,其主要药物动力学参数为:ρmax=(18.7±9.3)μg·L-1,tmax=(60.0±24.7)h,t1/2=(59.4±27.3)h,AUC0→t=(2 105.9±1 573.4)μg·h·L-1,AUC0→∞(2 297.1±1 734.7)μg·h·L-1。结论本方法适用于测定美洛昔康在人血浆中的浓度。     

HPLC法测定人血浆中盐酸二甲双胍的浓度及其在药动学研究中的应用

目的:建立了人血浆中盐酸二甲双胍浓度的反相离子对高效液相色谱测定法,研究复方盐酸二甲双胍片在中国男性健康志愿者体内盐酸二甲双胍的药动学行为,评价其生物利用度和生物等效性.方法:血浆样品经高氯酸沉淀蛋白,二氯甲烷脱脂后直接进样.流动相为甲醇-0.015mol·L-1磷酸二氢钾溶液(含0.001mol·L-1十二烷基磺酸钠,0.02mol·L-1氢氧化钾调节pH 5.2)(40:60),检测波长232nm,流速1mL·min-1.采用双交叉随机实验设计,20例受试者交叉口服复方盐酸二甲双胍试验片2片(每片含盐酸二甲双胍250mg和格列本脲1.25mg)或对照制剂盐酸二甲双胍片(每片含盐酸二甲双胍500mg)与格列本脲片(每片含格列本脲2.5mg)各1片,服药后0.5～24h内间隔取血.结果:HPLC测定盐酸二甲双胍在20～2 000μg·L-1范围内线性关系良好(r=0.999 8),最低定量限20μg·L-1,日内及日间精密度均小于10%.口服试验和对照制剂后,盐酸二甲双胍的t1/2,Tmax,Cmax分别为(3.84±0.61)h和(4.26±0.96)h,(2.0±0.7)h和(2.1±0.9)h,(1402.4±349.2)μg·L-1和(1 329.7±315.4)μg·L-1.以等剂量的盐酸二甲双胍与格列本脲片为对照,由AUC0～24计算,复方盐酸二甲双胍片中盐酸二甲双胍的相对生物利用度为(101.1±28.5)%.结论:本方法专属性强,灵敏度高.药动学试验结果表明试验复方片与联合使用的对照片盐酸二甲双胍生物等效.     

HPLC测定消渴丸中格列本脲的含量

目的制订消渴丸中格列本脲含量测定方法。方法采用高效液相色谱法测定格列本脲的含量。结果格列本脲在2.5μg～7.5μg范围内线性良好,平均回收率为95.69%,RSD为1.77%。结论含量测定方法简便易行,重现性好。     

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.     

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.     

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.