HPLC／MS测定人血浆中盐酸班布特罗及其代谢物特布他林的浓度

目的 :用高效液相 /质谱联用法测定人血浆中盐酸班布特罗及其代谢物特布他林的浓度。方法 :液相 :采用SupelcodiscoveryC18色谱柱 (5 μm ,2 5 0mm× 4 6mm) ;柱温 40℃ ;流动相为甲醇 -醋酸铵溶液 (0 0 0 7mol·L-1) (2 0∶80 ) (并用冰醋酸调 pH =4 8) ,流速 0 6mL·min-1,进样量 6 0 μL ;质谱 :大气压化学电离源 (APCI) ,选择性监测 (SIR)质荷比 (m/z)分别为 2 2 6 (特布他林 ) ,2 6 0 (内标 ) ,36 8(班布特罗 )带正电荷的分子离子峰定量。样品用固相萃取小柱提取处理。结果 :班布特罗线性范围 0 12 5～ 16 μg·L-1,最低检测浓度为 0 0 5 μg·L-1。特布他林线性范围 0 312 5～ 40 μg·L-1,最低检测浓度为 0 0 5 μg·L-1。班布特罗和特布他林的萃取回收率均在 90 %以上 ,日内、日间的RSD皆小于 15 %。结论 :适用于临床上测定血浆中盐酸班布特罗及其代谢物特布他林的浓度及药动学的研究     

HPLC/MS研究国产盐酸班布特罗片及其代谢物特布他林人体生物等效性

目的　研究国产班布特罗片剂和进口片剂进行人体生物等效性研究。方法　 2 0名健康受试者随机交叉给药 ,用液相色谱 /质谱联用测定血浆中班布特罗其代谢物特布他林的浓度。结果　经数据处理 ,单次口服国产和进口班布特罗片剂后班布特罗的药代动力学参数 :AUC0 -t分别为 (5 2± 2 1) μg·h·L-1和 (5 1± 2 0 ) μg·h·L-1,Tmax分别为 (2 9± 0 9)h和 (2 6± 0 7)h ,Cmax分别为 (6 0± 2 6 ) μg·L-1和 (6 2± 2 9) μg·L-1。特布他林 :AUC0 -t分别为 (191± 30 ) μg·h·L-1和 (197± 37) μg·h·L-1,Tmax分别为 (4 2± 1 0 )h和 (4 2± 1 0 )h ,Cmax分别为 (10± 5 )μg·L-1和 (10± 4) μg·L-1。国产班布特罗片剂单次给药后的相对生物利用度为 10 2 %± 8% (班布特罗 ) ,10 0 %±12 % (特布他林 )。结论　经统计学证明两制剂有生物等效性     

高效毛细管电泳法研究盐酸班布特罗国产胶囊与进口片剂的人体生物等效性高效毛细管电泳法研究盐酸班布特罗国产胶囊与进口片剂的人体生物等效性

目的研究国产盐酸班布特罗胶囊和进口片剂的人体生物等效性。方法采用高效毛细管电泳法测定血浆中班布特罗及其代谢物特布他林的浓度。结果单次口服国产班布特罗胶囊和进口班布特罗片剂后班布特罗的药代动力学参数:AUC0-t分别为(71±18)和(72±13) μg·h·L^-1,实测C_max分别为(8.1±1.8)和(9.2±2.3) μg·L^-1,实测t_max分别为(3.6±1.3)和(3.7±1.0) h。特布他林药代动力学参数:AUC0-t分别为(129±33)和(130±34) μg·h·L^-1,实测C_max分别为(7.8±2.3)和(8.5±2.9) μg·L^-1,实测t_max分别为(5.4±0.8)和(5.6±1.1) h,国产班布特罗胶囊单次给药后的相对生物利用度为(100±16)%(班布特罗),(101±13)%(特布他林)。结论经统计学证明两制剂具有生物等效性。     

高效毛细管电泳同时分析尿中20种碱性药物与毒物

目的 :建立用固相萃取高效毛细管电泳同时分离分析尿中 2 0种碱性药物与毒物的方法。方法 :尿样以oasis小柱提取 ,以多沙普伦为内标 ,采用未涂层毛细管 ( 75 μmID× 5 0 2cm ,有效分离长度为 4 0cm) ,缓冲液为 15 0mmol·L-1磷酸盐 + 2 0%甲醇 (pH =2 38) ,压力进样 8s,运行电压 16kV ,检测波长 2 0 0nm。结果 :本法分离效率高 ,药、毒物最低检测浓度为 15～ 80μg·L-1;线性范围 6 0～ 5 0 0 0 μg·L-1,日内、日间RSD <12 %。结论 :本法可用于多种类别的药、毒物的分析 ,增加检出几率 ,减少漏检。     

HPLC法同时测定血浆地西泮及其代谢物浓度

目的 :建立同时测定血浆中地西泮及其代谢物浓度的方法。方法 :选用ZORBAXRP C18柱 (15 0mm× 4 6mm ,5 μm) ;甲醇 - 2 5mmol·L-1醋酸铵溶液 (6 0∶4 0 ,V/V)作流动相 ;流速 0 8mL·min-1;检测波长 2 30nm。取血浆样品 0 5mL ,在碱性条件下用二氯甲烷 -正己烷提取 ,HPLC检测。结果 :本法对替马西泮、去甲地西泮和地西泮 3种物质的最低检测限均为 2 μg·L-1,线性范围为 10～ 15 0 0 μg·L-1;奥沙西泮的最低检测限为 5 μg·L-1,线性范围为 2 0～ 15 0 0 μg·L-1。回收率均接近 10 0 % ,日内、日间RSD <5 %。结论 :本法能同时测定血浆中地西泮及其代谢物浓度 ,具有重现性好 ,灵敏、可靠 ,可用于地西泮中毒的监测     

高效毛细管电泳法测定育亨宾树皮中育亨宾的含量

目的建立高效毛细管电泳(HPCE)测定育亨宾树皮中育亨宾含量的方法。方法运用高效毛细管电泳方法,熔融石英毛细管(75μmID×50cm),缓冲液为20mmol·L-1磷酸盐缓冲液(PH=3.0),检测波长:270nm,分离电压:15kV,柱温25℃,用0.45μm微孔滤膜过滤后进样,压力进样:50mbar×5s。结果方法最低检测浓度为0.1μg·mL-1,线性范围1～200μg·mL-1,r=0.9991,线性关系良好。精密度RSD为1.3%,回收率为98.4%。结论本法简单、灵敏经济,可作为育亨宾树皮中育亨宾的含量的测定。     

高效毛细管电泳法测定注射用头孢哌酮钠的含量

目的　测定注射用头孢哌酮钠的含量。方法　采用高效毛细管电泳法 ,毛细管 6 0cm× 75 μm ;运行缓冲液 30mmol·L-1硼砂溶液 (pH 9 2 ) ,高压进样 5s ,分离电压 12kV ,温度为 2 5℃ ,检测波长为 2 5 4nm ,地塞米松磷酸钠为内标。结果　头孢哌酮钠在 6～ 30 μg·ml-1浓度范围内线性关系良好 (r=0 9999) ,平均回收率为 99 17% (n =5 ,RSD =1.89% )。结论　所用方法简单、快捷、灵敏。     

高效毛细管电泳法测定氟尿嘧啶注射液中5-氟尿嘧啶的含量

目的建立高效毛细管电泳(HPEC)测定氟尿嘧啶注射液中氟尿嘧啶(5-FU)含量的方法.方法运用高效毛细管电泳方法,熔融石英毛细管(50 cm×57 cm,75 μm),缓冲液10 mmol·L-1磷酸二氢钠(pH 5.5),检测波长280 nm,分离电压15kV,用微孔滤膜过滤后进样,压力进样6 s.结果方法最低检测浓度为0.1 mg·L-1,线性范围5～200mg·L-1,r=0.999 0,线性关系良好.日内RSD为2.3%,日间RSD为2.7%,回收率为99.7%.结论本法简单、灵敏经济,可作为氟尿嘧啶注射液中氟尿嘧啶的含量的测定方法.     

槐角丸中染料木素的高效毛细管电泳测定

目的:建立适合槐角丸中染料木素测定的毛细管电泳方法。方法:采用高效毛细管电泳法。实验时,先分别用0.1 mol·L-1盐酸溶液、0.1 mol·L-1氢氧化钠溶液及运行缓冲液冲洗未涂层弹性石英毛细管(50μm ×43 cm)10 min,然后以槲皮素为内标,运行电压28 kV,运行缓冲液为20 mmo|·L-1磷酸盐缓冲液(pH 9.2),检测波长254 nm,室温下电动进样5 s。结果:染料木素的线性范围为6.25-31.26μg·mL-1(r=0.9994),其高、中、低3个量的加样回收率分别为100.7％,100.1％和99.2％,RSD分别为1.5％,0.63％和1.7％。结论:本法用于测定槐角丸中染料木素的含量快速、简便、准确可靠,有良好的重复性。     

氯霉素滴眼液及其相关物质二醇物的毛细管电泳分析法

目的　建立适合氯霉素滴眼液含量测定及其杂质二醇物检测的高效毛细管电泳定量检测方法。 方法 　采用毛细电泳法 ,以水杨酸为内标 ,运行电压 2 5 KV,运行缓冲液为 2 5 mmol· L- 1 磷酸盐缓冲液 (p H=6.0 ) ,检测波长 2 14 nm。结果 　氯霉素滴眼液及二醇物线性范围分别为 5 0～ 2 5 0 μg·ml- 1 (r=0 .9992 )和 4.0～ 2 0 .0 μg· ml- 1 (r=0 .9988)。平均回收率分别为 99.8%和 10 1.5 % ,RSD分别为 0 .8%和 2 .0 % (n=5 )。结论 　本方法能同时进行含量测定和杂质检测 ,对控制氯霉素滴眼液的质量有一定的价值     

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.