HPLC法测定体液中氨溴索浓度及其药代动力学参数

建立了 HPLC测定人血浆及尿中盐酸氨溴索含量的方法 :Hypersill C18柱 (4.6mm× 2 50 mm,5μm) ,乙腈 -甲醇 - 0 .0 1 mol/L磷酸盐缓冲液 -四氢呋喃 (35∶ 35∶ 2 7.5∶ 2 .5,V/V)为流动相 ,流速 1 .5ml/min,检测波长 2 4 2 nm。结果表明 :最低检测浓度为 5ng/ml,血药浓度在 1 0～ 32 0 ng/ml范围内线性良好 ,尿药浓度在 0 .2 5～ 8.0μg/ml范围内线性良好 ,氨溴索生物半衰期为 (4.2 1± 0 .93) h。     

人血浆中吡格列酮的固相萃取高效液相色谱法测定

以吡格列酮的同系物 PIOGA为内标 ,采用 C2 固相萃取紫外检测的 HPL C分析法测定人血浆中吡格列酮的浓度。 C1 8分析柱 (15 0× 4.6 mm,5 μm) ,流动相 :水 -乙腈 -冰醋酸 (5 40∶ 46 0∶ 1.2 ,用氨试液调至 p H6 .0 ) ,流速 :1.0 ml/ min,检测波长 2 6 9nm。取血样 0 .5 m l,加入内标经 C2 固相萃取后进样 40μl。最低定量限 (L OQ)为10 ng/ ml,线性范围为 10～ 16 0 0 ng/ ml     

3种交沙霉素片剂的生物等效性研究

目的 :研究 3种交沙霉素片剂在 12名男性健康受试者中的药物动力学和生物等效性。方法 :采用反相高效液相色谱法检测血浆中交沙霉素的浓度。结果 :交沙霉素的药 -时曲线经 3P97拟合符合开放性一室模型 ,A片的 AUC0 - t、AUC(0 -α) ,cmax、tmax、T1 /2 (ke) 分别为 (2 870 .75± 15 0 1.5 7) h· ng/ m l、(2 982 .0± 15 40 .91) h· ng/ m l、(115 8.2 8± 5 83.17) ng/ m l、(1.33± 0 .33) h、(1.18± 0 .40 ) h;B片分别为 (2 879.49± 136 5 .14) h· ng/ m l、(2 949.12± 140 4) h· ng/ ml、(130 8.40± 5 17.40 )ng/ ml、 (1.5 8± 0 .42 ) h、(1.19± 0 .2 3) h;C片分别为 (2 40 6 .77± 10 2 7.0 3) h· ng/ ml、 (2 482 .4± 10 2 8.2 8) h· ml、(1148.2 9± 784.0 3) ng/ m l、(1.5 8± 0 .19) h、(1.17± 0 .30 ) h。3种交沙霉素片剂的所有药物动力学参数经统计学 (SPSS软件的方差分析 )处理均无统计学差异 (P>0 .0 5 )。以片剂 B作为参比制剂 ,交沙霉素片剂 A和 C的生物利用度分别为 89.36 %和78.5 5 %。结论 :3种交沙霉素片剂具有生物等效性     

猪血浆中阿托品含量的毛细管气相色谱测定

采用气相色谱法测定猪血浆中阿托品的浓度。色谱柱为 HP- 5 (5 %苯基 - 95 %聚二甲基硅氧烷 )石英毛细管柱(30 m× 0 .5 3mm,1.5 μm) ,FID检测器。进样口温度 2 5 0°C,柱温 2 2 0°C,检测器温度 2 5 0°C。线性范围 0 .6 8～4 .2 5 μg/ m l(r=0 .9991) ,最低检测浓度 2 .5 ng/ ml     

维拉帕米及去甲基维拉帕米临床血药浓度的HPLC—FLU检测

目的 :建立一种简便的维拉帕米 ( Ver)及去甲基维拉帕米 ( Norv)血药浓度 HPL C- FL U的测定方法。方法 :色谱柱为Alltima C1 8柱 ( 15 0 mm× 4.6 m m,5 ︼m) ;流动相为甲醇 -水 -三乙胺 ( 5 2∶ 47.6∶ 0 .4) ,用冰醋酸调节 p H至 5 .41,流速 1ml/min。荧光检测激发波长 Ex=2 84nm,发射波长 Em=313nm。Ver和去 Norv的保留时间分别为 7.14min和 5 .17min,血样预处理采用乙腈直接沉淀法。结果 :Ver及 Norv血药浓度在 2 5～ 40 0 ng/ ml范围内线性关系良好 ( r≥ 0 .9997) ,最低检测限分别为 2 0 0 pg和 10 0 pg( r S/N≥ 3) ,最低检测浓度均为 2 0 ng/ ml。批内、批间精密度小于 5 %。结论 :本方法简便可靠 ,可较好地满足临床治疗药物监测工作的需要     

高效液相色谱法测定人血浆中阿糖胞苷的浓度

目的:建立以高效液相色谱法测定人血浆中阿糖胞苷浓度的方法。方法:色谱柱为Zorbax C18,检测波长为280nm,流动相为0·01mol/L磷酸盐(pH=7·0)-乙腈(95∶5),流速为1·0ml/min。结果:阿糖胞苷血药浓度在0·05~20μg/ml(r=0·9999)范围内线性关系良好,最低检测浓度为2ng/ml,方法回收率为98·3%~103·2%,日内、日间相对标准差分别为2·8%、3·6%。结论:本方法简单、快速、准确,适用于阿糖胞苷临床血药浓度监测和药动学研究。     

高效液相色谱法测定人血浆中盐酸氨溴索浓度

目的 :建立测定盐酸氨溴索血药浓度的方法。方法 :采用高效液相色谱法 ,以盐酸地尔硫为内标 ,色谱柱采用HypersilBDSC18 不锈钢柱 (250mm×4 6mm ,5μm ) ,流动相为乙腈 -甲醇 -0 01mol/L磷酸盐缓冲液 (pH7 0) -四氢呋喃 (35∶35∶27 5∶2 5) ,流速为1 0ml/min ,检测波长为242nm。9名健康男性志愿者 ,单剂量口服进口盐酸氨溴索片后 ,测定其血浆中药物浓度。结果 :线性范围为10～640ng/ml,最低检测浓度为10ng/ml ,提取回收率均大于90 %。9名志愿者的血药浓度数据经3p87拟合 ,符合血管外给药一室模型 ,Cmax= (226 53±34 59)ng/ml ,Tmax= (1 82±0 80)h ,T1/2ke= (6 27±0 66)h ,AUC0～24=(2363 55±448 86) (ng·h)/ml。结论 :本方法简单 ,灵敏度和准确度较高 ,能满足人体药动学研究的需要。     

RP-HPLC荧光法测定兔血浆中秦皮甲素的浓度

目的建立测定兔血浆中秦皮甲素浓度的RP-HPLC-FLD方法。方法以C18柱(150mm×4.6mm,5μm)分离;流动相为甲醇-水(76∶24);流速:1ml.min-1;柱温:30℃;荧光检测器,激发波长230nm,发射波长460nm。结果秦皮甲素的保留时间为(3.620±0.05)min;血药浓度在0.1～100mg·L-1范围内线性关系良好(r=0.997);绝对回收率为74.0%～82.6%,日内、日间精密度(RSD)均<9.0%,最低检测限为0.05ng。兔单次静脉注射秦皮甲素5mg·kg-1后,T12a、T21β分别为(0.196±0.008).h-1,(6.70±0.645).h-1。结论该法快速、灵敏、准确,可用于秦皮甲素的体内分析。     

吡喃霉素在乳腺癌患者体内的药代动力学

目的 :建立HPLC分离和检测人血浆中吡喃霉素 (Pir)的分析方法 ,并研究Pir在乳腺癌患者体内的药代动力学。方法 :以正定霉素为内标 ,1ml血浆加0 1ml(0 4mol/L ,pH=9 0)的氯化铵缓冲液碱化后以氯仿 -甲醇 (2∶1 ,v/v)混合溶剂提取2次。在Phe nomenex(C18)柱上 ,以0 04mol/L磷酸二氢钾 (pH=3 0)∶乙腈=68∶32为流动相 ,荧光检测波长480nm/550nm(Ex/Em )。结果 :血浆中Pir的线性范围5～1000ng/ml(r=0 9997) ,方法回收率为95 3 % ,日内及日间变异性RSD均小于5 % ;Pir在人体的药代动力学特征为开放性二室模型 ,消除相半衰期T1/2β、清除率CLs、表观分布容积Vd及曲线下面积AUC分别依次为 (12 8±5 9)h、(128 3±52 6)L/(m2·h)、(1754 3±478 2)L/m2 和 (428 7±137 2)ng/(h·ml)。结论 :本方法适用于临床Pir的血药浓度监测和药代动力学研究。     

HPLC法测定盐酸二甲双胍的血药浓度

目的 :用HPLC测定血浆中盐酸二甲双胍。方法 :采用LichrospherC1 8柱 ( 4.6× 2 5 0mm ,5 μm)为色谱柱、3mmol/L十二烷基磺酸钠水溶液 (含 0 .5 %三乙胺 ,滤过后调 pH =4) -乙腈 ( 3 5 /65 )为流动相 ;流速为 1ml/min ,检测波长为 2 3 2nm。结果 :标准曲线的线形范围为 0 .0 5～ 4μg/ml(r=0 .9997,n =7) ,最低检测浓度为 5 0ng/ml,低、中、高 3种浓度的日内和日间变异分别为 5 .2 4% ,2 .41 % ,1 .3 0 % (n =5 )和 8.0 4% ,4.65 % ,4.3 4% (n =5 ) ,相对回收率分别为 96.1 7% ,94.0 3 %和 1 0 4.2 2 % ,绝对回收率分别为 ( 85 .47± 5 .1 4) % ,( 92 .0 3± 2 .3 4) % ,( 92 .63± 2 .0 3 ) %。结论 :本方法简便、快速、准确、专一性好 ,可用于盐酸二甲双胍的血药浓度测定     

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.