HPLC法测定全血中环孢素浓度

目的建立测定全血中环孢素(CsA)浓度的固相萃取HPLC法。方法全血样品用硫酸锌甲醇混合液处理,取上清液进样C18(10 mm×3.9 mm)柱,经水冲洗去杂质后,用纯甲醇洗脱CsA,进入C18(250 mm×4.2 mm)柱,以流动相:乙腈-甲醇-水(60∶10∶30,V/V/V)洗脱后进行定量测定。结果本法最低检出浓度为40μg.L-1,线性范围为62.5~2 000μg.L-1,方法回收率为90%~95%,提取回收率>80%(n=5),日内RSD为3.25%~4.12%,日间RSD为3.37%~6.87%。结论本法测定CsA前处理步骤简便,结果可靠,可用于临床标本检测。     

高效液相色谱法测定甲氨蝶呤的血药浓度

目的:建立以高效液相色谱法测定人血清中甲氨蝶呤(MTX)浓度的方法。方法:采用Shim-pack VP-ODS(150mm×4.6mm,5μm)的色谱柱;流动相:乙腈-0.3%磷酸(15∶85,用三乙胺调pH2.5);紫外检测波长为302nm;柱温:40℃;流速:0.8mL.min-1。结果:MTX在0.1~100.0mg.L-1质量浓度内线性关系良好,Y=0.2+30 174X,r=0.999 4;MTX低、中、高(0.25,12.50,100.00mg.L-1)3种质量浓度的平均回收率分别为104.0%,102.3%,102.5%。日内、日间RSD均小于3%,最低血清检测浓度50μg.L-1。结论:高效液相色谱法测定MTX的血药浓度灵敏度高,结果准确可靠,操作简便,能满足其血清浓度的测定。     

高效液相色谱法测定硝西泮的血药浓度

目的测定血清中硝西泮浓度;方法采用高效液相色谱法,甲基睾丸酮为内标,经YW G-C18色谱柱(4.6mm i.d.×200 mm,5μm)分离,流动相为〔乙腈-甲醇(1∶2,v/v)〕-水(58∶42,v/v),检测波长为265nm。结果在45.4~454μg.L-1浓度范围内硝西泮和内标的色谱峰高比与浓度呈线性关系(r=0.9999);平均回收率为92.08%(RSD=2.30%,n=9);最低检测限为25μg.L-1;结论本法简便、准确,适用于临床上对硝西泮的血药浓度的常规监测。     

甲氨蝶呤血药浓度监测方法的改进及其临床应用

目的建立改进的甲氨蝶呤(MTX)血药浓度监测方法,为临床合理应用MTX提供参考。方法采用高效液相色谱外标法,色谱柱:Aglient TC-C18柱(4.6×250mm,5μm),流动相:0.025mol·L-1磷酸二氢钠溶液(pH 5.41)-甲醇(76∶24,V/V),流速:1.0mL·min-1,检测波长:313nm,柱温:35℃,样品经高氯酸沉淀蛋白后直接进样。结果血浆MTX浓度在0.05～5.00μg·mL-1和5.00～80.00μg·mL-1范围内线性关系良好,r1=0.9999,r2=0.9992,平均回收率分别为98.74%和98.60%,平均提取回收率为69.43%和70.99%,最低定量浓度为0.05μg·mL-1,日内及日间RSD均小于4%。结论改进后的方法简便快速、准确可靠,灵敏度高,重现性好,线性范围宽,成本低,适用于甲氨蝶呤血药浓度监测。     

恶性淋巴瘤患者血清甲氨蝶呤的浓度监测

目的建立反相高效液相色谱法监测恶性淋巴瘤患者血清中甲氨蝶呤(MTX)浓度,为临床合理用药提供依据。方法色谱柱HWG-C18(4.6mm×200mm,5μm),柱温:35℃,流动相为甲醇-磷酸盐缓冲液(19∶81,V/V),检测波长:306nm,流速:1.0mL.min-1,血浆样品经高氯酸沉淀蛋白后进样20μL。结果血清MTX浓度在0.01~10.00和10.00~100.00μg.mL-1范围内线性良好,平均回收率为92.00%,日内及日间RSD均<6.00%。结论该法简便、准确、灵敏,适用于应用大剂量MTX患者血药浓度的监测。     

HPLC法测定人血浆中西洛他唑的浓度

目的建立测定人血浆中西洛他唑浓度的HPLC法。方法以迪马公司钻石C18反相柱(250 mm×4.6 mm,5μm)为色谱柱,流动相为甲醇-0.03 mol.L-1醋酸铵(62∶38,V/V),流速0.8 mL.min-1,检测波长257 nm,以乙酸乙酯为提取剂。结果西洛他唑高(1 000μg.L-1)、中(500μg.L-1)、低(25μg.L-1)3种质量浓度的平均回收率分别为99.51%、97.07%、98.28%,日内、日间RSD均<7%;分析方法的最低检测浓度为10μg.L-1。线性范围为10~1 500μg.L-1。结论该方法灵敏、准确、简单、快速,可用于西洛他唑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氯丙嗪的浓度

目的建立测定人血浆中氯丙嗪浓度的高效液相色谱法。方法以DiamonsilTM C18反相柱(150mm×4.6mm,5μm)为色谱柱,流动相0.03mol·L-1醋酸铵-甲醇(21∶79);流速:1.0mL·min-1;柱温:40℃;检测波长:254nm。结果氯丙嗪血药浓度在10.0~1000.0μg·L-1范围内,与峰面积比有良好的线性关系(r=0.9994)。其10.0,400.0,1000.0μg·L-13种浓度平均回收率分别为100.16%,98.33%,97.57%;分析方法的最低检测浓度为10.0μg·L-1;日内、日间差RSD均低于8%(n=5)。结论本法灵敏、准确、简单、快速,可用于临床血药浓度监测和药动学研究。     

HPLC法测定人血浆中替硝唑的浓度

目的采用HPLC法测定人血浆中替硝唑的浓度。方法以D iam onsilTMC18反相柱(250 mm×4.6 mm,5μm)为色谱柱,流动相为甲醇-0.03 m o l.L-1醋酸铵(25∶75,V/V),流速0.8 mL.m i-n 1,紫外检测波长320 nm,以乙酸乙酯∶二氯甲烷(4∶1,V/V)为提取剂。结果替硝唑的高、中、低3种浓度的回收率分别为111.01%、99.00%和100.74%,日内RSD≤5.87%(n=5)、日间RSD≤7.82%(n=3);线性范围为0.25~75 m g.L-1,回归方程为:c=25.65F-0.03,r=0.9998(n=9);最低检测浓度为0.05 m g.L-1(R sn≥3)。结论该方法可用于临床血药浓度监测和药动学研究。     

高效液相色谱法同时测定血清中普罗帕酮和小檗碱的浓度

目的:建立同时测定血清普罗帕酮和小檗碱浓度的高效液相色谱法.方法:采用YWG-C18柱(5 mm×150 mm),流动相为甲醇-(醋酸钠-醋酸缓冲液)-二乙胺-水(68∶25∶2∶5),紫外波长240 nm检测.结果:本法对普罗帕酮和小檗碱最低有效检测浓度均为0.05 mg·L-1;回收率接近100%,日内、日间精密度＜5%;线性范围均为0.05～5.00mm mg·L-1.结论:方法简单、灵敏、准确,可用于临床药物监测.     

高效液相色谱法测定血清中氯唑沙宗的浓度

目的建立测定血清中氯唑沙宗浓度的高效液相色谱法。方法采用迪马C18色谱柱(250×4.6mm,5μm);流动相:乙腈-0.02MKH2PO4(pH6.3)=9∶11(V/V),含0.16%三乙胺;流速:1.0mL.min-1;紫外检测波长:280nm;柱温:30℃。结果氯唑沙宗在2.5~12.5μg.mL-1的浓度范围内线性关系良好,最低检测限为0.4μg.mL-1,回收率为102.82±4.39%。结论本法操作简便,灵敏度高,快速可靠,可用于血清中氯唑沙宗的浓度测定。     

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.

---

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.