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单击以编辑,母版标题样式,单击以编辑母版文本样式,第二级,第三级,第四级,第五级,#,药物分析药物现代仪器分析法课件,1,第十六章 药物现代仪器分析法,药物现代仪器分析法,1.,色谱法,2.,光谱法,3.,质谱法,(MS),4.,毛细管电泳法,5.,联用技术,6.,微型全分析系统,(Lab-on-a-chip),7.,场流分离技术,(FFF),第十六章 药物现代仪器分析法药物现代仪器分析法1.色谱法2,2,1.,色谱法,色谱法,1.,薄层色谱法,(TLC),:定性鉴别、杂质检查,2.,毛细管气相色谱法,(GC),残留溶剂的检测,药物的定量分析。,3.,高效液相色谱法,(HPLC),药物含量测定,手性药物拆分。,色谱系统适用性试验,:,柱效,(,即理论塔板数,n),分离度,R (,一般,R,1.5,),拖尾因子,T ( 0.95,1.05,),重复性,(,一般,RSD,2.0%,),1.色谱法色谱法1. 薄层色谱法(TLC):定性鉴别、杂质,3,2.,光谱法,紫外,可见,光谱法,(UV) , A,=ECL,灵敏度达,10,-4, 10,-6,g/ml,波长(,200 400, 760 nm,),2.,荧光光度法,(,检测光与入射光成直角,), F=KC,灵敏度达,10,-7, 10,-10,g/ml,一般采用,激发光波长,250 700 nm,4.,红外光谱法,(IR),主要用于有机药物的定性鉴别 波长,400 760 nm,5.,核磁共振法,(NMR),,主要用于有机药物的定性鉴别,6. X,射线粉末衍射法,主要用于药物晶型的定性鉴别,3.,原子吸收光度法主要用于金属元素的分析,灵敏度达,10,-6, 10,-9,g/ml,现代光谱法,2. 光谱法紫外可见光谱法 (UV) , A=ECL,4,3.,质谱法,(MS),原理:,将化合物形成离子和碎片离子,按其质荷比的不同进行分离,可做成分和结构分析。,特点:,应用范围广、灵敏度高,(10,-11,g),、分析速度快,(,小于,1,秒,),。,样品导入系统,离子源,质量分析器,检测器,记录仪,用途:,测定分子量,鉴定化合物,推测未知物结构,测定分子中,Cl,和,Br,等的原子数,如与色谱联用可进行多组分的定性定量分析。,3. 质谱法(MS)原理:将化合物形成离子和碎片离子,按其,5,4.,高效毛细管电泳法,(High Performance Capillary Electrophoresis, HPCE),原理:,依据样品中各个组分之间淌度和分配行为上的差异实现分离分析。以石英毛细管为分离通道、高压直流电场为驱动力。,优点:,高效,(n,可达几十万至几千万,),高速,(,最快可在,1,分钟内完成,一般几分钟即可,),微量,(nL,级,即,10,-9,L),低消耗,(,流动相 几,mL,即可,),分离模式:,毛细管区带电泳,(CZE),胶束电动毛细管色谱,(MECC),毛细管凝胶电泳,(CGE),毛细管等电聚焦,(CIEF),4. 高效毛细管电泳法 原理:依据样品中各个组分之间淌度和,6,药物分析药物现代仪器分析法课件,7,药物分析药物现代仪器分析法课件,8,5.,联用技术,1. GC IR,:中成药复方制剂、 药用挥发油、香精香料分析、毒物检测、废水和农药分析等,2. GC MS,:合成产物的确证、副产物的鉴定、中药未知成分的鉴定、药物代谢的研究。,LOQ,可达,10,-9, 10,-12,g,3. HPLC MS,:药品质量控制,(,杂质、降解产物、生物转化产物等,),体内药物分析,药物代谢研究,4. HPLC NMR,:药物杂质鉴定、药物体内外代谢产物的结构鉴定、天然产物化学筛选等,5. HPCE MS,:合成产物的确证、副产物的鉴定、中药未知成分的鉴定,couple,5. 联用技术1. GC IR:中成药复方制剂、 药用,9,6.,微型全分析系统,(Miniaturized Total Chemical Analysis System, -TAS),1990,年左右,,Manz,和,Widmer,率先提出微全分析系统,(Miniaturized total analysis system,TAS),,全世界分析化学家高度重视,,2001,年新的学术季刊,“,Lab-on-a-chip,”,创立,美国,Aanalytical Chemistry,期刊每两年的综述中列有专门的文章综述,TAS,领域的发展。,TAS,领域中微加工方法,大致以最简易方法为主。,TAS,检测器:激光诱导萤光检测器;吸收光度检测器;化学发光检测器;折光率检测器;发射光谱检测器;电化学检测器;质谱检测器,等,。,TAS,卡脖子难点:,1),进样系统,2),前处理不容易,3),重复性差,TAS,的巨大潜力:,1),多重平行检测,2),大规模集成化,未来的研究领域:,与生命科学相关,如生物医学、高通量的药物合成筛选、卫生检疫等。随着人类基因组计划的初步完成,已步入后基因组时代,单核甘酸多态性分析、基因表达分析、基因变异分析以及蛋白组分析等, 由于,TAS,有大规模平行处理能力,可望成为后基因组时代的支撑性技术。, 1990年左右, Manz 和Widmer 率先提,10,药物分析药物现代仪器分析法课件,11,药物分析药物现代仪器分析法课件,12,药物分析药物现代仪器分析法课件,13,7.,场流分离技术,F,ield-flow fractionation (FFF) is the most versatile separation technique of all macro molecular and particle separation methodologies in terms of separation range, selectivity and resolution. FFF comprises a family of techniques that have demonstrated, over more than two decades, the ability to characterize supra molecular species in a size range spanning,many order of magnitude, from macromolecules to micron-sized particles.,The separation occurs by differential retention in a stream of liquid flowing through a thin, empty channel. The separated components are eluted one at a time into the detector.As in classical chromatography, sample components elute at given,retention times,which are related, and often rigorously predictable, to various physicochemical properties of the retained species. Measurements of retention times can yield these properties for each fractionated component.This is the main distinguishing feature of FFF with respect to classical chromatography that can neither directly predict retention for a given component nor extract chemical information straight from retention parameters.,7. 场流分离技术 Field-flow f,14,The FFF mechanism combines elements of chromatography and field-driven techniques such a electrophoresis and ultracentrifugation. Like chromatography, FFF is an elution technique; like field-driven techniques, FFF requires a field or gradient. The field in FFF is applied at,right angles,to flow and serves to drive components into different stream laminae in a capillary channel. The different velocities of the fluid laminae across the channel develops the separation induced by the action of field.,The FFF Family,Sedimentation FFF,FlowFFF,ThermalFFF,Steric FFF,What can you play with FFF,?,SOME APPLICATIONS,Proteins, protein aggregates,lipoproteins,DNA, viruses, chromosomes,Pharmaceutical emulsions, liposomes,Bacterial cells, blood cells,The FFF mechanism combines ele,15,What do you need to work with FFF,What do you need to work with,16,FFF differs from a standard HPLC in the column. Here the column is the channel. The channel is capillary with rectangular cross-section. There is no sta,t,ionary phase inside the channel and separation is generated by an external field applied perpendicularly to the mobile phase flowing inside the channel.An FFF system can, therefore, be easily implemented in labs with a standard HPLC expertize.,Thank you for your attenation!,FFF differs from a standard HP,17,
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