RP-HPLC法测定大鼠血浆中3-正丁基苯酞的含量及其药动学

目的建立大鼠口服及静脉注射3-正丁基苯酞(NBP)的药动学研究方法。方法以雄性Wistar大鼠为实验对象,分别采取静脉注射和口服给药2种方式,于给药后不同时间点取血,采用高效液相反相色谱检测法(RP-HPLC)测定大鼠血浆中NBP的含量,选用Hypersil ODS色谱柱(250 mm×4.6 mm,5μm),以0.05 mol·L~(-1)醋酸钠溶液-乙睛(50:50,V/V)为流动相,流速为1.0 mL·min~(-1),检测波长为228 nm。应用3P97药动学软件处理血药浓度数据,获得药动学参数。结果 NBP在0.050 38～21.23μg·mL~(-1)(r=0.999 7)范围内线性关系良好,其中定量下限为0.050 38μg·mL~(-1);日内和日间精密度均小于4.8%,低、中、高浓度的NBP提取回收率均在80%以上。NBP药动学过程符合双隔室开放模型。大鼠静脉注射NBP 5、10、20 mg·kg~(-1)的主要药动学参数分别为t_(1/2)(2.67±0.72)h、(2.23±0.92)h、(2.27±0.52)h;AUCo_(0-t)(8.15±1.46)mg·h·L~(-1)、(7.23±1.55)mg·h·L~(-1)、(14.05±1.67)mg·h·L~(-1)。口服60、240和360 mg·kg~(-1)NBP的主要药动学参数依次为P_(max)(2.90±0.94)μg·mL~(-1)、(5.46±0.56)μg·mL~(-1)、(9.46±1.61)μg·mL~(-1);AUC_(0-t)(7.11±2.90)mg·h·L~(-1)、(14.00±0.82)mg·h·L~(-1)、(24.33±2.86)mg·h·L~(-1)。结论本法灵敏度高、专属性强、易操作,结果准确可靠,可用于NBP的药动学研究。     

液相色谱-串联质谱法测定老年人血浆中石杉碱甲的浓度

目的建立测定老年人血浆中石杉碱甲的液相色谱-串联质谱方法。方法在血浆中加入内标石杉碱乙,用乙腈直接沉淀蛋白法处理样品。采用SeQuant ZIC-HILIC(100mm×2.1mm I.D,3.5μm)色谱柱进行分离,柱温35℃;流动相为含5 mmol·L~(-1)甲酸铵0.1%甲酸水溶液-乙腈(35:65,V/V),流速0.3 mL·min~(-1),柱温35℃,梯度洗脱,进样体积5μL;正离子多反应检测(MRM),离子通道分别为m/z 243.2→210.2(石杉碱甲),m/z 257.2→198.6(石杉碱乙)。结果石杉碱甲线性范围0.05～10μg·L~(-1)(r>0.999),定量下限0.05μg·L~(-1),提取回收率在81.0%～95.4%,批内、批间精密度RSD均小于8%,内标校正基质因子在2.11～2.28。主要药动学参数t_(max)为(2.2±0.7)h,ρ_(max)为(0.9±0.2)μg·L~(-1),t_(1/2)为(14.1±2.0)h,AUC_(0-∞)为(15.3±4.5)μg·h·L~(-1)。结论本方法操作简便、特异性强、灵敏度高,适用于石杉碱甲在老年人中的药动学研究。     

肝损害大鼠ig泛普洛韦的药代动力学

目的:研究肝损害大鼠ig泛昔洛韦(famciclovir)后体内喷昔洛韦(penciclovir)药代动力学,探讨肝损害大鼠肝醛氧合酶体外氧化泛昔洛韦的酶动力学行为。方法:用HPLC法测定CCI_4引起肝损害的SD大鼠ig泛昔洛韦200mg·kg~(-1)后的血浆中喷昔洛韦浓度及肝损害的SD大鼠肝醛氧合酶匀浆中泛昔洛韦浓度。结果:肝损害大鼠喷昔洛韦药代动力学参数为t_(1/2ka)(0.34±0.13)h,C_(max)(43.4±27.6)mg·L~(-1),AUC_(0～8h)106.3±19.5mg·L~(-1)·h[对照分别为(0.53±0.14)h,(27.6±6.4)mg·L~(-1),(91.8±17.3)mg·L~(-1)·h,P<0.05]。肝损害大鼠肝醛氧合酶体外氧化泛昔洛韦的酶动力学参数为K_m(0.1632±0.0882)mmol·L~(-1)、V_(max)(36.87 15.46)nmol·min~(-1)·mg~(-1)protein[对照组K_m,V_(max)分别为(0.1184±0.0479)mmol·L~(-1),(51.72±21.09)nmol·min~(-1)·mg~(-1)protein;P<0.05和P<0.01]。结论:肝损害大鼠对泛昔洛韦吸收和转化成喷昔洛韦能力明显减低,但消除基本不变。肝醛氧合酶的亲和力和最大反应速率均降低,可能是肝损后泛昔洛韦药动学行为改变的因素之一。     

LC-MS法测定人血浆中米氮平浓度以及药代动力学研究

目的:建立人血浆中米氮平浓度的HPLC—MS分析方法,用以测定健康受试者口服米氮平制剂后的血药浓度,估算受试制剂和参比制剂的药代动力学参数,评价两种制剂的生物等效性和相对生物利用度。方法:血浆中加入内标地西泮后,环己烷提取,HPLC—MS分离、分析。色谱系统:Shimadzu ODS柱(150 mm×4.6 mm),甲醇-醋酸铵溶液(0.01 mol·L~(-1))(75:25)为流动相,流速1.0 mL·min~(-1),柱温30℃。质谱检测方式:SIM。结果:米氮平的线性范围为0.2—120 ng·mL~(-1)(r=0.9992),最低检测浓度达0.1 ng·mL~(-1),提取回收率≥85%。测定20名健康志愿者单剂量交叉口服试验制剂与参比制剂后的血药浓度经时过程,受试制剂和参比制剂的主要药代动力学参数:C_(max)(71.53±10.5)ng·mL~(-1)和(73.48±17.8)ng·mL~(-1);T_(max)(2.0±0.8)h和(1.9±0.7)h;t_(1/2)(22.33±2.05)h和(22.97±3.56)h,无显著性差异;受试制剂的相对生物利用度为(101.3±16.2)%。结论:本法专属、准确、灵敏。统计学结果表明受试制剂和参比制剂生物等效。     

HPLC法测定人血浆中比阿培南的浓度及其药动学研究

目的:建立一种简便、快速测定人血浆中比阿培南浓度的方法,并考察其药动学。方法:血浆样品加入5-羟基吲哚乙酸20μL,经7%硫酸锌蛋白沉淀后进样测定,其中色谱柱为AgilentZorbaxBonus-RP,流动相为甲醇-0.3%醋酸水溶液(梯度洗脱),流速为1.0mL·min~(-1),紫外检测波长为300nm,进样量20μL。结果:30名健康受试者随机分为低、中、高3个剂量组单次静脉滴注,血浆中比阿培南的药动学参数分别为:t_(1/2)(1.13±0.16)、(1.06±0.10)、(1.11±0.12)h,t_(max)(1.02±0.21)、(1.10±0.31)、(1.05±0.27)h,c_(max)(9.04±4.24)、(17.64±4.55)、(35.10±4.23)μg·mL~(-1),AUC_(0～12h)(18.17±3.86)、(36.44±7.68)、(70.24±8.91)mg·h·L~(-1),AUC_(0～∞)(20.42±4.69)、(37.21±7.63)、(72.40±9.03)mg·h·L~(-1);多次给药的t_(1/2)、t_(max)、c_(max)、AUC_(0～12h)、AUC_(0～∞)分别为(1.07±1.35)h、(1.04±0.60)h、(19.61±3.68)μg·h~(-1)、(32.47±3.90)mg·h·L~(-1)、(33.36±3.80)mg·h·L~(-1)。结论:3个剂量组的药动学参数呈线性相关,各剂量组内、间无显著性个体差异,不同年龄、性别、身高、体重之间也无显著性个体差异。     

高效液相色谱-质谱法测定大鼠血浆中硫酸酯型脱氢表雄酮和孕烯醇酮

建立同时测定 SD 大鼠血浆中脱氢表雄酮硫酸酯(DHEAS)和孕烯醇酮硫酸酯(PREGS)的高效液相色谱-质谱法。方法:选择雌酮硫酸酯(ES)作为内标。雄性 SD 大鼠的血浆使用乙腈沉淀除去蛋白后,经固相萃取、水解及衍生化后,以高效液相色谱质谱检测器测定脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯的浓度。色谱柱为 Zorbax SB C_(18)柱温40℃,流动相为乙腈水,使用梯度洗脱;采用大气压化学离子源(HPLC/APCI-MS),负离子检测方式,用于选择离子检测(SM)定量分析的离子为 m/z 490.0(DHEAS)和 m/z 518.0(PREGS)及 m/z 472.1(ES)[M-H]~-。结果:血浆中脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯线性范围分别为:0.05～2.0ng·mL~(-1),0.03～2.0 ng·mL~(-1),r 分别为0.9990和0.9979。低、中、高浓度血样的提取回收率在66.8%～82.3%之间,相对回收率为:98.4%～111.6%,日内、日间变异均小于15%。正常雄性 SD 大鼠的血浆脱氢表雄酮硫酸酯和孕烯醇酮硫酸酯分别为0.070±0.020 ng·mL~(-1)、0.13±0.031 ng·mL~(-1)。结论:本实验方法具有灵敏度高、准确度好及变异较小的特点,适合测定雄性 SD 大鼠血浆中脱氢表雄酮硫酸酯和和孕烯醇酮硫酸酯的含量。     

大鼠血浆中木兰花碱的HPLC法测定及药动学研究

目的:建立大鼠血浆中木兰花碱的高效液相色谱分析方法,研究其在大鼠体内药动学过程。方法:以塔斯品碱为内标,采用甲基叔丁基醚沉淀蛋白。色谱柱为Agilent C18(250mm×4.6mm,5μm),流动相为甲醇-0.05mol·L-1 KH2PO4-三乙胺(42:58:0.5,V:V:V),流速为1.0mL·min-1,紫外检测波长为260nm。结果:大鼠血浆内源性物质对木兰花碱测定无干扰,木兰花碱检测浓度在0.50～30.00μg·mL-1范围内与木兰花碱对内标塔斯品碱的峰面积之比线性关系良好(r=0.9994);提取回收率≥91.05%,方法回收率在90.57%～95.38%之间,日内、日间精密度RSD均≤8.42%(n=6)。木兰花碱在大鼠体内的血药浓度-时间曲线符合二室模型。结论:该方法准确、简便、灵敏,适用于木兰花碱在大鼠体内的药动学研究。     

清胰Ⅱ号颗粒在大鼠体内药动学研究

目的:研究大鼠ig给予清胰Ⅱ号颗粒,其有效成分大黄素在大鼠体内的药动学过程。方法:大鼠分别ig给予不同剂量(2.5、5.0 g·kg~(-1))的清胰Ⅱ号颗粒,采用高效液相色谱法在不同时间点测定血浆中大黄素浓度,并计算药动学参数。结果:给予不同剂量药物的大鼠血浆中大黄素的t_(1/2α)分别为(9.468±8.46)、(21.68±17.867)h;t_(1/2β)分别为(15.388±5.46)、(39.63±24.39)h;t_(max)分别为(2.500±3.479)、(5.333±3.266)h;C_(max)分别为(0.058±0.004)、(0.101±0.007)mg·L~(-1);CL分别为(33.027±9.365)、(9.405±5.846)L·h~(-1)·kg~(-1);AUC(0-∞)分别为(0.652±0.201)、(1.364±0.267)mg·h~(-1)·L~(-1),其动力学过程符合二室模型。结论:建立了大鼠血浆中大黄素浓度检测方法,得到了相关药动学参数,可为清胰Ⅱ号颗粒后续药动学-药效学研究奠定理论基础。     

灯盏花乙素苷元不同给药剂量的大鼠药动学比较

目的:比较灯盏花乙素苷元不同剂量给药的大鼠药动学。方法:12只大鼠随机均分为2组,分别单剂量灌胃给予灯盏花乙素苷元在20和200 mg·kg~(-1),用高效液相-电化学色谱法(HPLC-ECD)测定给药后不同时间血浆中灯盏花乙素苷元及其代谢物灯盏花乙素的浓度,计算AUC_(0-24h)。色谱柱为Kromasil C_(18)柱(150 mm×4．6 mm,5μm);流动相为pH 2．6的50 mmol·L~(-1)磷酸盐缓冲液-甲醇-四氢呋喃(60:40:10);流速为1．0 mL·min~(-1);分析电压为100 mV。结果:灯盏花乙素苷元20 mg·kg~(-1)剂量给药时,原型药完全被代谢,血浆中只检测到代谢产物灯盏花乙素;200 mg·kg~(-1)剂量给药时,血浆中代谢产物灯盏花乙素和原型药灯盏花乙素苷元共存。灯盏花乙素苷元200 mg·kg~(-1)剂量时,代谢产物灯盏花乙素的AUC值相仅较20 mg·kg~(-1)剂量时提高3倍(1934．83 vs 536．51 ng·mL~(-1)·h)。结论:大小剂量下灯盏花乙素苷元的药动学行为呈非线性。     

左氧氟沙星对黄芩苷血药浓度的影响

目的:研究口服抗生素药物左氧氟沙星对中药黄芩苷血药浓度的影响。方法:用HPLC-ECD方法测定左氧氟沙星(30 mg·kg~(-1),po,bid,给药4 d)和黄芩苷(34 mg·kg~(-1),po,给药一次)合并用药组与黄芩苷(34 mg·kg~(-1),po,给药一次)单独用药组大鼠在给予黄芩苷后不同时间的血浆黄芩苷浓度,比较两者的药动学参数。结果:合并用药组中黄芩苷的C_(max)=(485．73±217．57)ng·mL~(-1),AUC_(0-24h)=(5508．49±2374．16)ng·mL~(-1)·h;单独用药组中黄芩苷的C_(max)=(2645．62+601．42)ng·mL~(-1),AUC_(0-24h)=(28952．90±5731．42)ng·mL~(-1)·h。合并用药组的C_(max)和AUC_(0-24h)显著低于单独用药组(P＜0．05)。结论:左氧氟沙星严重影响黄芩苷的血药浓度,提醒临床医生和患者应合理用药。     

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.