反相高效液相色谱法测定人血清中格列齐特浓度

目的 :建立快速分离与测定人血清中格列齐特的反相高效液相色谱法。方法 :固定相为ZorbaxODS柱 ,以甲醇 0 .2 %冰醋酸 (5∶4,v/v)为流动相 ,流速 1.0ml·min-1,检测波长 2 2 9nm ,格列齐特的保留时间为 7.92min。结果 :本法在 0 .0 1～10 .0 0 μg·ml-1间线性良好 ,相关系数r =0 .9997。高中低浓度样品的日内和日间测定相对标准偏差RSD均小于 5 % ,平均回收率为 (10 1.1± 4.8) % ,最低检测血清浓度为 0 .0 1μg·ml-1。结论 :本方法可用于格列齐特的临床药物动力学研究及临床血药浓度监测     

HPLC测定人体血浆中替硝唑浓度

目的:建立替硝唑血药浓度测定方法,用于复方替硝唑片血药浓度研究。方法:采用高效液相色谱法。色谱柱为C18柱,流动相为12%乙腈水溶液(pH7.0),检测波长318nm。结果:样品组分在0.2～20μg.ml-1线性良好(r=0.9998),最低血浆药物检测浓度为0.02μg.ml-1。回收率为94.4%～98.3%,日内及日间RSD<5%。结论:本法简便、迅速、灵敏、准确,适合于临床药学研究需要。     

复合氨基酸胶囊中水溶性维生素的含量测定

目的:建立复合氨基酸胶囊中5种水溶性维生素的含量测定方法.方法:采用高效液相色谱法,用乙腈与庚烷磺酸钠-磷酸钠的水溶液(pH=3.0)为流动相,流速1.0ml·min-1,色谱柱为C18(4.6×250mm,5μm),柱温40℃,检测波长为280nm,进样量为20μl.结果:维生素Bl、维生素B2、维生素B6、维生素C和烟酰胺分别在24.7～75.2μg·ml-1、15.0～44.9μg·ml-1、12.2～37.5μg·ml-1、99.2～299.9μg·ml-1和100.3～300.9μg·ml-1范围内呈线性关系,相关系数均为1.000,平均回收率均在100%±2%范围以内.RSD分别为0.19%、0.26%、0.46%、1.17%和0.52%.结论:本方法准确,简便,快捷,可以同时测定复合氨基酸胶囊中维生素B1、维生素B2、维生素B6、维生素C和烟酰胺的含量.     

RP-HPLC测定人血浆中的阿昔洛韦

目的采用RP-HPLC法测定人血浆中的阿昔洛韦。方法Lichrospher C18色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-10 mmol.L-1磷酸二氢钠缓冲液(3:97,磷酸调pH3.4);流速1.0 ml.min-1;检测波长254 nm;柱温40℃。结果阿昔洛韦在0.05~5.00μg.ml-1线性良好(r=0.9994),最低检测浓度为50.00 ng.ml-1,方法回收率93.33%~104.96%,日内、日间RSD分别为1.46%~3.83%和2.32%~6.37%。结论所建方法简便、快速、灵敏、准确、干扰少,适用于阿昔洛韦血药浓度的测定。     

高效液相色谱法测定阿昔洛韦尿素乳膏中阿昔洛韦的含量

建立阿昔洛韦尿素乳膏中阿昔洛韦含量的HPLC测定方法.色谱柱为SUNTEK Kromasil C18;流动相为甲醇-水(10:90),流速为1.0ml·min-1,检测波长为254nm.线性范围:2.5～20μg·ml-1,r=0.9999 ,平均回收率为96.90%,RSD为0.77%.方法准确可靠,简单易行,适用于阿昔洛韦尿素乳膏中阿昔洛韦的含量测定.     

SPE-HPLC测定索他洛尔血药浓度

目的:建立检测索他洛尔血药浓度的SPE-HPLC方法.方法:以C18固相萃取小柱预处理样品.色谱柱为Shim-pack VP-ODS C18柱,流动相为乙腈-水-三乙胺(5:95:0.2,v/v,磷酸调pH4.0),流速0.8 ml·min-1,柱温30℃,检测波长为227 nm,进样量20μl.结果:索他洛尔在0.04～5.0μg·ml-1浓度范围内线性关系良好,回归方程为Y=1.79X-0.02,r=0.999 8,索他洛尔萃取回收率为91.05%～100.75%,方法回收率为98.55%～101.36%(n=5),日内RSD小于5%,日间RSD小于10%.结论:本方法灵敏、准确,可用于索他洛尔的血药浓度检测.     

RP-HPLC法测定大剂量化疗时人血浆中甲氨喋呤浓度

目的建立大剂量化疗时人血浆中甲氨喋呤浓度的测定方法。方法采用反相高效液相色谱法(RP-HPLC)。色谱柱:μBondapakC18(4.6×150mm),柱温:35℃,流动相为甲醇∶(pH7.2)磷酸缓冲液(17∶83,v/v),流速:1.0ml·min-1,检测波长306nm。结果线性范围0.08～100μg·ml-1,线性关系良好,r=0.9999,最低检测浓度0.01μg·ml-1,回收率102.0%,日内及日间RSD均<5%。结论该方法简单快捷、灵敏度高、结果准确,适用于大剂量化疗时甲氨喋呤血药浓度监测。     

RP-HPLC法测定维生素B1,B2及B6片中含量

目的:建立一种反相高效液相色谱法测定,维生素B1、维生素B2、维生素B6的含量测定方法.方法:Hypersil, ODSC18柱(250mm×4.6mm,5μm),流动相:甲醇-50mmol·L-1磷酸二氢钾溶液(20:80),检测波长为266nm,流速1.0ml·min-1.结果:维生素B1、维生素B2、维生素B6;线性范围分别为:16.05～80.25μg·ml-1(r=0.9999)21.35～106.75μg·ml-1(r=0.9999)及14.15～70.75μg·ml-1(r=0.9999),平均回收率也分别为101.2%,98.7%及99.6%,RSD 1.9%,1.9%及1.0%.结论:该方法准确,灵敏度高,重现性好,可作为质量检验的定量方法.     

布洛芬颗粒剂的人体生物等效性研究

目的:研究布洛芬颗粒剂的人体生物等效性.方法:HPLC法测定18名男性健康志愿者单剂量口服400 mg布洛芬颗粒剂后的血药浓度,拟合药动学参数并进行统计学评价.结果:单剂量口服布洛芬颗粒剂试验制剂和参比制剂后,血浆中布洛芬的Cmax别为(39.35±5.42)μg·ml-1和(40.10±6.33)μg·ml-1;tmax分别为(1.64±0.41)h和(1.58±0.19)h;AUC(0-10)分别为(142.70±25.02)μg·ml-1·h和(150.29±18.24)μg·ml-1·h;AUC(0-in分别为(152.93±28.14)μg·ml-1·h和(161.18±19.59)μg·ml-1·h.相对生物利用度为(95.97±18.91)%.结论:经方差分析与双单侧t检验,试验制剂与参比制剂具有生物等效性.     

尼美舒利颗粒剂的人体生物等效性及相对生物利用度研究

目的 建立人血浆中尼美舒利的固相萃取结合HPLC-UV测定法,研究两种尼美舒利颗粒的生物等效性.方法 采用XB-Phenyl柱(250mm×4.6mm,5μm),柱温为40℃,流动相为乙腈-30mM乙酸铵(44:56,v/v),流速为1.0ml·min-1,检测波长为350nm.18例男性健康志愿者自身对照随机交叉给药,分别单次口服不同厂家尼美舒利颗粒剂200mg,不同时间点取血,比较两药主要药动学参数的差异和相对生物利用度.结果 尼美舒利与血浆中内源性杂质分离完全,尼美舒利在0.092～15.300μg·ml-1内与峰面积比线性良好,血浆中尼美舒利最低定量浓度为0.092μg·ml-1.方法 的回收率为92.2%～102.2%,日内、日间精密度(RSD)均<8.0%.单剂量口服尼美舒利200mg后,两种制剂的AUC0→t为(83.51±23.34)μg·h-1·ml-1和(86.54±27.92)μg·h-1·ml-1;AUC0→∞为(87.00±26.23)μg·h-1·ml-1和(91.04±32.57)μg·h-1·ml-1;Cmax为(10.18±1.76)μg·ml-1和(10.60±1.62)μg·ml-1;tmax为(2.1±0.7)h和(2.0±0.8)h.结论 该方法简单快速,灵敏度高,可用于尼美舒利的体内过程研究.方差分析表明,两制剂之间药动学参数无显著差异,试验制剂与参比制剂为生物等效制剂.     

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.     

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.     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

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.