HPLC柱前衍生化法测定硫酸依替米星的含量

目的 :采用反相高效液相色谱柱前衍生化法测定硫酸依替米星的含量。方法 :衍生化条件 :衍生化试剂为邻苯二醛 ,反应温度为 6 0℃ ,反应时间为 15min。色谱条件 :色谱柱为KromasilC1 8柱 (5 μm ,2 0 0mm× 4 6mm) ;流动相为水 冰醋酸 甲醇 (2 0∶5∶75 )配制的 0 0 2mol L的庚烷磺酸钠溶液 ;检测波长为 330nm ;流速为 1ml min。结果 :线性范围 :0 1～ 1 0mg ml,r=0 9999;该方法与微生物法测定结果基本一致 ,衍生化产物稳定。结论 :所建立的反相高效液相色谱柱前衍生化法能准确测定硫酸依替米星的含量 ,重复性好     

高效液相色谱法测定卡马西平片的含量

建立卡马西平片含量测定的HPLC方法。色谱柱为LichrosorbDiol (4 0× 2 5 0mm ,5 μm ) ;流动相 :乙腈 -甲醇 - 0 0 5 %冰醋酸溶液 (5∶5∶90 ) ;流速 :1 0ml·min-1;检测波长 :2 30nm ;柱温 :2 5℃ ;峰面积外标法。卡马西平在 10～ 10 0 μg·ml-1范围内具有良好的线性关系 ,回归方程为A =1 6 95 3× 10 4C - 2 5 5 5 5× 10 3 ,r=0 99997;平均回收率为 99 6 9% ,RSD =0 75 % (n =6 )。本法准确可靠 ,专属性强 ,能更好地控制其质量。     

HPLC法测定艾司奥美拉唑钠中3个潜在基因毒性杂质的研究及控制

目的:建立HPLC法测定艾司奥美拉唑钠中潜在基因毒性杂质E、杂质I、2-氯甲基-3,5二甲基-4-甲氧基吡啶。方法:杂质E色谱条件：YMC-Triart C₁₈色谱柱(250 mm×4.6 mm, 5μm),流动相A为0.05 mol·L^-1磷酸二氢钾缓冲液,流动相B为乙腈,梯度洗脱,流速为1.0 mL·min^-1,检测波长为302 nm,柱温为30℃;杂质I色谱条件：Agilent Microspher C₁₈色谱柱(100 mm×4.6 mm, 3μm),流动相A为水-磷酸盐缓冲液(pH 7.6)-乙腈(80∶10∶10),流动相B为乙腈-磷酸盐缓冲液(pH 7.6)-水(80∶1∶19),梯度洗脱,流速为1.0 mL·min^-1,检测波长为302 nm,柱温为30℃;2-氯甲基-3,5二甲基-4-甲氧基吡啶色谱条件：GL Inertsil ODS-3色谱柱(250 mm×4.6 mm, 5μm),流动相A为0.01 mol·L^-1磷酸氢二钠溶液(pH...     

雷丸中16种氨基酸的柱前衍生化RP-HPLC法含量测定

目的:测定雷丸中16种氨基酸的含量。方法:样品以6 mol.L-1盐酸于150℃水解1 h,以异硫氰酸苯酯(PITC)为衍生化试剂衍生后进行HPLC分析。采用Phenomenex Luna C18(250 mm×4.6 mm,5μm)色谱柱,柱温为43℃;流动相A为0.1mol.L-1醋酸钠缓冲液(以醋酸调pH 6.5)-乙腈(93∶7),流动相B为乙腈-水(4∶1),梯度洗脱,流速为1.0 mL.min-1;检测波长为254 nm。结果:16种氨基酸浓度在1.16～47.32μg.mL-1范围内呈良好的线性关系(r=0.9998～1.000);平均回收率在93.3%～107.2%之间;RSD均小于3.0%。结论:本方法灵敏、准确,具有良好的重复性和稳定性,可用于雷丸中氨基酸的检测。     

氨基酸分析法测定复方骨肽注射液中多肽含量

目的建立复方骨肽注射液中多肽含量的氨基酸分析法。方法氨基酸水解方法:6mol.L-1盐酸110℃水解22h;氨基酸衍生方法:异硫氰酸苯酯(PITC)法;HPLC条件:Agilent Eclipse AAA色谱柱;流动相A:0.1mol.L-1乙酸钠(用冰醋酸调至pH 6.5)-乙腈(97∶3);流动相B:乙腈-水(4∶1);流速:1.0mL.min-1;检测波长:254nm。结果 18种氨基酸的线性、精密度及稳定性良好;除水解样品中的胱氨酸与色氨酸外,其他氨基酸的回收率均达到定量要求。结论方法准确、专属性强,可作为复方骨肽注射液中多肽的定量分析方法。     

高效液相色谱法测定复方盐酸二甲双胍片中盐酸二甲双胍及格列本脲含量

目的测定复方盐酸二甲双胍中盐酸二甲双胍及格列本脲的含量。方法优化高效液相色谱法条件。 (1)盐酸二甲双胍的色谱条件是 :色谱柱 :HypersilCN(2 0 0mm× 4 6mm ,5 μm,大连依利特 ) ;流动相 :乙腈 - 0 0 0 5mol·L-1NH4H2 PO4(pH 3 5 ,5 3∶4 7) ;检测波长 :2 34nm ;柱温 :35℃ ;流速 :1 0mL·min-1。 (2 )格列本脲的测定 :色谱柱 :HypersilODS柱 (2 0 0mm× 4 6mm ,5 μm,大连依利特 ) ;流动相 :甲醇 - 0 0 0 5mol·L-1NH4H2 PO4(pH3 5 ,5∶3) ;检测波长 :30 0nm ;柱温 :35℃ ;流速 :1 0mL·min-1。结果盐酸二甲双胍和格列本脲的线性范围分别为 5 0～ 5 0 0mg·L-1和4 1 2～ 16 4 8mg·L-1;RSD均不超过 1% ;盐酸二甲双胍质量分别为 80 1、10 0 2、12 0 7μg的回收率分别为 99 9%、10 0 1%、10 0 0 % ,格列本脲质量分别为 80 2 4、10 0 30、12 0 36 μg的回收率分别为 10 1 0 %、99 7%、10 0 2 % ;最低检测限分别为 0 2 0和 0 80mg·L-1。结论本法适于复方盐酸二甲双胍的含量测定     

HPLC法测定帕尼培南/倍他米隆的含量

建立了HPLC法测定帕尼培南 /倍他米隆的含量。色谱条件 :采用C18柱 (5 μm ,2 0 0mm×4 6mm) ,0 0 2mol/L磷酸盐缓冲液 (pH8 0 )—乙腈 (97∶3 )为流动相 ,柱温 45℃ ,检测波长 2 60nm。帕尼培南和倍他米隆分别在 119～ 478和 12 6～ 5 0 5 μg/ml范围内呈良好的线性关系 ,日内RSD分别为 0 4%和 0 5 % ,回收率分别为 99 8%和 10 0 0 % ,含量测定结果与进口药品注册标准方法所得结果相符。     

高效液相色谱法测定人血浆中两性霉素B浓度

目的　测定人血浆中两性霉素B浓度。方法　色谱柱为SupelcosilLC 18 DB( 4 .5mm× 2 5 0mm ,5 μm) ,流动相为0 .0 2 5mol/L乙二胺四乙酸二钠 (EDTA 2Na) 乙腈 ( 5 0∶5 0 ) ,柱温 :2 5℃ ,流速 1.0mL/min ,检测波长 4 0 5nm。结果　人血浆中两性霉素B浓度在 0 .0 5～ 4 .0 μg/mL范围内线性关系良好 ,回归方程为 :C =5 .0 4× 10 -6A + 0 .0 7(r =0 .9994 )。高、中、低 3个浓度平均回收率为 ( 98.4± 6 .9) % ;日内RSD≤ 3.5 % ,日间RSD≤ 7.6 %。结论　本法灵敏、快速、重现性好 ,分析时间短 ,可用于临床药代动力学研究及血药浓度监测     

高效液相色谱-蒸发光散射检测法测定盐酸大观霉素的含量

目的 :应用高效液相色谱 -蒸发光散射检测 (HPL C/ EL SD)法分析盐酸大观霉素含量。方法 :采用 Diamonsl TM(钻石 ) C1 8色谱柱 (2 5 0 mm× 4 .6 mm,5μm) ;流动相 :0 .0 2 mol/ L三氟醋酸水溶液 -乙腈 (99:1) ;检测器条件 ,漂移管温度为 5 0℃ ,雾化气体 (空气 )压力为 3.0 bar。结果 :盐酸大观霉素在 2 .72 5～ 10 .90 4μg范围内 ,进样量对数值与峰面积对数值呈良好的线性关系 ;理论板数以大观霉素峰计算为 5 10 0。结论 :本方法操作简单 ,准确性、重现性好 ,与微生物效价法比较 ,测定大观霉素含量无显著性差异 ,可用于盐酸大观霉素成品的质量控制。     

甲磺酸罗哌卡因中2,6-二甲基苯胺的HPLC-电化学法测定

建立了测定甲磺酸罗哌卡因中 2 ,6 -二甲基苯胺的高效液相色谱 -电化学检测法。采用 C1 8柱 ,流动相为含乙二胺四乙酸二钠和磷酸氢四丁铵的 0 .5 mol/ L磷酸盐缓冲液 -乙腈 (6 0∶ 4 0 ) ,流速 1.0 m l/ min,电极电压为 0 .90 V。2 ,6 -二甲基苯胺在 5× 10 - 3～ 6× 10 - 2 μg/ ml的范围内线性良好 (r=0 .9980 ) ,RSD为 2 .7%。     

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.