RP-HPLC测定消痔灵片中胆酸的含量

目的采用反相液相色谱-紫外检测法测定消痔灵片中胆酸的含量.方法采用十八烷基键合硅胶色谱柱(Hypersil C18,150 mm × 4.6 mm, 5 μm).流动相:乙腈-水-冰醋酸(37 ∶63∶0.01),流速:0.8 mL·min-1,检测波长190 nm,外标法定量.结果消痔灵片中胆酸与其他成分得到较好分离,在0.05 ～ 4.00 mg·mL-1范围内线性良好(r = 0.9999);回归方程为 Y = 11.99+2086.02 X.检测限为0.9 μg ;样品低,中,高3种浓度的方法回收率为 98.4 % ～99.7 %;精密度 RSD < 2.0 %.结论本法结果准确,重复性良好,方法简便可行.     

危重症感染患者血清中替考拉宁浓度测定

目的:建立高效液相色谱法快速测定人血清中替考拉宁浓度的方法,并用于危重症感染患者治疗药物监测。方法:色谱柱:Hypersil C18柱(250mm×4.6mm,5μm);流动相:0.01mol.L-1磷酸二氢钠-乙腈-甲醇(70∶25∶5,pH2.1),流速1.0mL.min-1。紫外检测波长为240nm,进样量20μL,柱温25℃。结果:替考拉宁在5.63~125.00mg.L-1内线性关系良好(r=0.999 5),最低定量限为5.63mg.L-1,提取回收率为95.13%,方法回收率为98.13%,日内、日间精密度的RSD分别为3.2%和6.8%。另外,所监测的8例肾功能不全患者中,有50%(4/8)患者替考拉宁质量浓度小于10mg.L-1。结论:HPLC简便、灵敏、快速、重复性好,可用于危重症感染患者替考拉宁的血药浓度监测;危重症感染患者应用替考拉宁后血药浓度变化较大,需要通过治疗药物监测的方法制定个体化给药方案。     

高效液相色谱法测定三七伤药片中芍药苷含量

目的:用HPLC法测定三七伤药片中芍药苷的含量.方法:色谱柱为ODS-C18;乙腈-0.1%磷酸溶液(17∶83)为流动相;检测波长为230 nm.结果:芍药苷在10.84～54.20 mg·L-1范围内呈良好线性关系(r=0.999 3),精密度高,RSD为0.7%.平均回收率为99.1%,RSD为1.9%(n=5).结论:方法简便可行,重现性好,可作为本制剂的质量控制标准.     

高效液相色谱法测定盐酸吡格列酮的血药浓度

目的:建立高效液相色谱法测定人血浆中盐酸吡格列酮浓度的方法.方法:以卡马西平为内标,血浆样品用醋酸乙酯萃取,采用Diamonsil(TM)C18柱(250 mm×4.6mm,5μm)分析.流动相为乙腈-0.05 mol·L-1磷酸二氢钾(pH 6.5)(42:58),流速1.2 mL·min-1,检测波长229 nm,柱温35℃.结果:本法在0.025～4.0 mg·L-1间线性关系良好,γ=0.999 9,RSD为2.08%,(n=6),最低检测质量浓度为0.02 mg·L-1.日内、日间精密度RSD均小于10%,低、中、高浓度的提取回收率均大于99%(n=5).结论:此方法灵敏度高、重复性好,尤其适用于临床治疗合并用药时的药物浓度监测.     

HPLC法同时监测5种抗精神病药血药浓度

目的:建立以高效液相色谱法同时监测氯丙嗪、氯氮平、氯硝西泮、地西泮、阿普唑仑血药浓度的方法。方法:用乙酸乙酯提取血清中的药物,并进样分析,其中色谱柱为NovapakC18,流动相为甲醇-乙腈-水-三乙胺-冰醋酸(16∶36.5∶47.5∶0.03∶0.02),检测波长为254nm,流速为1.5mL.min-1,柱温为50℃,灵敏度为0.01AUFS。结果:血清中5种药物检测浓度均在0.03～100μg·mL-1范围内线性关系良好,方法回收率为88.6%～98.7%,日内和日间精密度小于5.0%。结论:本方法简便、快速、灵敏、准确、用血量少,可用于临床用药监护以及急性药物中毒快速定性和定量分析。     

梯度洗脱高效液相色谱法检测注射用替加环素的有关物质

目的建立测定注射用替加环素有关物质的梯度洗脱反相高效液相色谱法。方法采用Wa-ters C18色谱柱(5μm,416 mm×250 mm),以磷酸盐缓冲液-乙腈(95∶5)为流动相A,以磷酸盐缓冲液-乙腈(50∶50)为流动相B。梯度洗脱条件:0~40 min,A:85%→57%;40~55 min,A:57%→0%;55~56 min,A:0%→85%;以1.0 mL/min的流速进行梯度洗脱,检测波长为248 nm。结果在上述色谱条件下,替加环素与各中间体杂质及降解杂质均能有效分离,分离度大于2.0;检测限为3.6 ng,精密度良好(RSD为0.7%)。结论本方法操作简便,专属性强,灵敏度高,可用于注射用替加环素有关物质的测定。     

HPLC-UV法测定人血浆及尿液中比阿培南的浓度

目的建立HPLC-UV法测定人血浆及尿液中比阿培南的浓度。方法血浆样品经超滤离心法提取后,采用XB-C18柱分离,流动相:乙腈-0.1 mol.L-1醋酸铵缓冲液(pH 4.6)=3∶97(v/v),流速:1.0 mL.min-1,检测波长:300 nm,柱温:40℃,进样量:30μL;尿样处理方法同血样,但以甲醇代替乙腈,进样量20μL。结果比阿培南与血浆、尿液中内源性物质分离度好,血药、尿药浓度分别在0.1~50、0.5~300 mg.L-1与峰面积线性关系良好,定量下限分别为0.1、0.5 mg.L-1,绝对回收率分别在90.9%~97.3%、99.1%~102.5%,相对回收率分别为102.1%~110.1%、100.3%~104.4%,日内精密度(RSD)分别<3.4%、2.1%,日间精密度(RSD)分别<5.7%、2.8%。均满足生物样本测定要求。结论所建立方法高效、灵敏、简捷快速、专属性好、重现性高,可用于比阿培南药动学研究。     

离心分配色谱法测定阿昔洛韦乳膏中主成分的含量

目的:建立测定阿昔洛韦乳膏中主成分含量的方法。方法:采用离心分配色谱法。溶剂体系为正己烷-乙腈-水(2∶1∶1,V/V/V),进样溶剂为5%吐温80水溶液,进样量为1.0 ml,流速为5 ml/min,检测波长为254 nm。结果:阿昔洛韦质量浓度在0.012 7~0.126 7 mg/ml范围内与峰面积呈良好的线性关系(r=0.998 7);精密度、稳定性、重复性试验的RSD≤2.0%;平均加样回收率为97.34%,RSD为0.90%(n=9)。结论:该方法精密度好、准确度高,可用于测定阿昔洛韦乳膏中主成分的含量。     

高效液相色谱法测定人脑脊液中利奈唑胺浓度

目的:建立高效液相色谱法快速测定人脑脊液中利奈唑胺浓度并对危重患者进行药物浓度监测。方法:采用外标法,色谱柱:ZORBAX Edipse XDB-C18(4.6 mm×150 mm,5μm);流动相:乙腈-水(23∶77);流速:1.0mL·min-1;柱温:30℃;紫外检测波长:254 nm,并利用建立的方法对危重患者进行利奈唑胺脑脊液浓度的监测。结果:利奈唑胺在0.31～40 mg.L-1范围内线性关系良好(r=0.999 9),最低定量限为0.31 mg.L-1,平均绝对回收率为46.8%,平均相对回收率为96.99%,高、中、低3个浓度日内、日间精密度的RSD均小于<5%。另外,所监测的危重患者中,利奈唑胺不同时期在脑脊液中的浓度变化很大。结论:所建立的高效液相色谱法简单、快捷、灵敏、准确,可用于监测利奈唑胺在脑脊液中的浓度。     

用柱前衍生RP-HPLC法测定脑蛋白水解物注射液中牛磺酸的含量

目的:建立测定脑蛋白水解物注射液中牛磺酸含量的柱前衍生RP-HPLC法。方法:在碱性条件下,氨基酸与异硫氰酸苯酯反应,生成异硫氰酸苯?氨基酸的衍生物,采用Diamonsil C18色谱柱(250 mm×4.6 mm,5μm),用二元梯度洗脱方式,流动相A为0.1 mol/L乙酸钠?乙腈(93∶7),流动相B为乙腈?水(80∶20),检测波长254 nm。结果:牛磺酸在0.01~0.07 mg/mL浓度范围内呈良好的线性关系(r=0.999 9),精密度良好,低、中、高3种浓度(0.019、0.024、0.029 mg/mL)的加样回收率分别为(97.0±0.5)%、(98.8±0.20)%、(100.0±1.6)%(n=3)。结论:本法结果准确,重复性好。     

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.     

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.