第八章质谱法.ppt

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,*,第八章 质谱法,Mass Spectroscopy(MS),质谱仪的一般结构,真空系统,进样系统,离子检测器,数据系统,质量分析器,离子源,大气,样品,机械泵，分子涡轮泵，扩散泵，,离子泵，升华泵,气体：气相色谱仪,(,GC/MS),液体：液相色谱仪,(,LC/MS),固体：固体进样杆,(,DIP,DCI),电子倍增器，多通道板检测器,光电倍增管，阵列检测器,通常的离子化方法,电子轰击离子化,EI,化学电离离子化,CI,正化学电离离子化,PCI,负化学电离离子化,NCI,大气压力离子化,API,电喷雾离子化,ESI,大气压化学电离离子化,APCI,GC/MS,LC/MS,离子化方式,分子量,非极性,极性,EI/CI,ESI,APCI,100,000,1000,离子化和碎片,离子化,硬,无碎片,有碎片,API,EI,CI,软,质量分析器,四极杆,80%,的台式质谱仪(3000/年,),HP,PE,TMQ,Shimadzu,离子阱,20%,的台式质谱仪,(700/,年,),Varian,TMQ,飞行时间(,ToF,),目前用户较少(50-60/年),Leco,Micromass,TMQ,扇形磁场,通常不是台式仪器,四极质量分析器,+,+,+,+,离子阱,原理与四极杆的非常相似，离子被存储在,RF,和,DC,场中，扫描场逐出精确,m/z,的离子。,飞行时间质量分析器,+,+,+,+,所有离子同时启动,小的离子移动速度快,测量已知距离下的速度,(,飞行时间),扇形磁场质量分析器,+,+,+,离子通过变化的磁场和电场被传输,Electron Impact Ionization(EI),e,-,e,-,e,-,e,-,e,-,Sample,from GC,filament,+,+,+,+,Fragment ion,QP,Electron Impact Ionization(EI),Thermal electrons(ca.70,eV,)hit molecules and ionize them.,Ample fragment ions=ample structural information Finger print,EI spectrum,Methylstearate,M.W.298,Positive Chemical Ionization(PCI),from,gas cylinder,filament,e,-,e,-,e,-,e,-,e,-,QP,CH,4,CH,5,+,C,2,H,5,+,Sample,from GC,H,+,-CH,4,M+1,-C,2,H,4,Positive Chemical Ionization(PCI),Reagent gas molecules are ionized first.,Sample molecules are ionized mainly,by,損,roton,transfer?,Quasi molecular ions easily produced,useful for molecular weight verification.,+,PCI spectrum,Methylstearate,M.W.,298,Comparison of EI and PCI,EI mode,PCI mode,Fragment,Molecular weight,Negative Chemical Ionization(NCI),Negative ion formation,by electron capture,associative resonance capture,dissociative,resonance capture,ion-pair production,Negative ion formation,by ion-molecule reactions,proton transfer,charge exchange,nucleophilic,addition,nucleophilic,displacement,Negative ion formation by electron capture (associative resonance capture),MX+e,-,(0,eV,)MX,-.,X:positive electron affinity part,CH4,e,-,MX,CH4,+,CH3,+,H,CH4,CH4,CH4,e,-,MX,-,质谱图的样子?,EI,和,CI,的质谱图,50,100,150,200,250,300,350,400,450,500,550,m/z,0,100,%,510,511,512,50,100,150,200,250,300,350,400,450,500,550,m/z,0,100,%,241,213,510,242,EI,O,O,O,C,F,3,C,F,3C,H,3,NCI,扫描,50,到300,amu,流入质谱仪的化合物,质谱图.,60,80,100,120,140,160,180,200,220,240,260,280,300,0,100,%,240,241,断裂机理,1.,断裂,2.,断裂,3.,i,断裂,4.逆,Diels,-Alder,反应,5.,Mclafferty,重排,Configuration of GCMS,Mass Spectrometer,Gas Chromatograph,Interface,Ion source box,Ion Source Design,-Increased ionization efficacyInterface,The sample separated,by GC will be introduced,through this interface.,Filament,Ion generated by filament will give the electron impact to the sample to produce the sample ionization.,Ion source,box,Ion Lens,-High ion introduction to mass analyzer-,This ion lens is functioning for effective introduction,to mass analyzer.Optimization for adjustment of,this ion lens will be done by PC automatically.,Quadrupole,Rod,-Minimize losses in transit to the,Quadrupole,-,Main Rod,Specific ion only can be passed by the electronic field,into,Quadrupole,Rod which is controlled by PC.,Pre Rod,Fromion source,To,detector,rotary pump,turbo molecular pump(151 L/sec),turbo molecular,pump(151 L/sec),rotary pump,Good vacuum system,-Minimize losses in transit to the,Quadrupole,-,Turbo Molecular Pump,Air cooled and oil free,Turbomolecular,Pump(TMP)with lubricant impregnated ceramic bearings is maintenance free for up to 25,000 hours.,Provides high quality vacuum in less than 5,mins,.,Ion detection,+7.5,kV,-,+,e,-,-0.73.0,kV,AMP,-7.5,KV,+,e,-,-0.73.0,kV,AMP,e,-,Negative ion detection,Positive ion detection,GCMS-QP5050A NCI,sensitivity,200,fg Hexachlorbenzene,NCI spectrum,Mass chromatogram,of m/z284,OFN(,octafluoronaphthalene,)NCI scan mode,Outstanding High Sensitivity,scan range,m/z 60-310,0.5sec interval,200,fg,S/N 30,GCMS-QP5050A,NCI pesticides date 3,10,pg,Cyanophos,NCI,mass spectrum,phosphorus pesticides 10pg,Comparison between EI and NCI,extract from cabbage(pesticides added),EI scanning mode,NCI scanning mode,Comparison between EI and NCI,extract from cabbage(pesticides added),Principle of,Programed,temperature vaporize(PTV),PTV,100,C,split,MS,50-250,C,Solvent will be,vaporized and,exhausted.,Sample will be,concentrated.,Optimization of PTV conditions,Selection of packing materials,Tenax,GC,20%OV-1,20%OV-17,20%,Dexil,300,Shincarbon,A,Carbopack,C,Trap temperature,Desorption,temperature,Temperature program,Very-Large-Volume Injection using PTV,Effect of Temperature on Trap efficiency,Very-Large-Volume Injection using PTV,Temperature and Split ratio Program,Very-Large-Volume Injection using PTV,2ppb each Pesticides in n-hexane,1000,uL,Injection,Analytical ConditionsGC17A Ver.2DB1 30mX0.25mm50C(4)-160C-280C 20C/min 5C/minPTV70C(1)-100C(1)-280C 30C/min 250C/minWide range FTD 1pA 280CCarrier gas He 150kPa,Precolumn Tenax,GC,Features of GCMS QP-5050A,Outstanding high sensitivity,Extended mass range up to 900,amu,Versatile Mode-EI/PCI/NCI/DI,Compact Design,Easy Operation using Windows Software,Easy tuning-PFTBA built in standard,Easy maintenance,Analysis of Coconut oil,MS Spectrum of C48,DI option,The direct inlet option allows less volatile or thermally labile samples to be directly introduced into the mass spectrometer.Mixtures of compounds which have different boiling points can be positively identified by programming the DI probe temperature.,Application using direct inlet probe,Front-open ion chamber system,-easy maintenance-,Dual Filaments System,-easy maintenance-,LCMS-QP8000,Validation,Productivity and Mass Detection,What kind of benefits LC/MS users can get?,Powerful qualitative capability,Selective quantitative capability,M/Z,M/Z,M/Z,What kind of benefits LC/MS users can get?,Powerful qualitative capability,Selective quantitative capability,A:100,D:150,B:100,C:150,m/z=150,TIC,m/z=100,A,B,C,D,HPLC,MS,New Shimadzu LCMSQP-8000,QP-8000 is LC/MS with API interface,which is designed to be a routine LC detector,in our new VP series HPLC system.,LCMS Interface,Curved,Desolvation,Line,(CDL),Deflector,Probe,Spray Chamber,Skimmer,Lens Stack,RP TMP1 TMP2,Quadru,pole,API Spray Chamber,API Spray Chamber,ESI Probe(Easy to change to APCI Probe),API Probe,ESI and APCI probes are included as standard.They can be applied to various kinds of samples.All you have to do to switch the ionization method is to change the prove.,APCI probe,ESI Probe,Mobile phase,LC flow rate and,Nebulizer,flow rate slightly influence spray condition.,You can adjust the probe position to the best for each condition.,The APCI probe contains,corona discharge needle,.So no need to change the API spray chamber.,Easy Maintenance,Skimmer Lens stack access,Spray Chamber Access,You can pick it up from upper side without any tools.,You can remove the spray chamber from the front side.,It will be easy to wash this part after analysis.,Standard Syringe Pump,Syringe for,direct sample,injection,Syringe for standard sample,of calibration,LCMS Application,Mass Separation,Column:STR ODS-II(2.0mmI.D.x150mm),Mobile phase:65%,acetonitrile,-water containing 0.1%formic acid,0.2ml/min,Maltose,TIC,387,225,195,Glucose,Xylose,Analysis of,macrolide,antibiotics,UV 210nm,MDM,SIM,SPM-I,II,SPM-III,OM,DH EM,MDM,Summary,LCMS-QP8000 is a,Compact Bench Top,Liquid Chromatograph with high performances,Easy Operation and Maintenance,using CLASS-8000 software,Self diagnostic functions,User login and security defined by access levels.,OLE custom report generation,Affordable price,质谱图和色谱图,?,如果我们注意这个区域.,色谱图?,1.25,1.50,1.75,2.00,2.25,2.50,2.75,3.00,3.25,Time,50,100,%,TIC,在此的细微观察是由许多点组成的,.,色谱图-放大,2.12,2.14,2.16,2.18,2.20,2.22,2.24,2.26,2.28,Time,78,100,%,TIC,每个点相对于质量分析器的一个扫描,(,质谱图,).,60,80,100,120,140,160,180,200,220,240,260,280,300,0,100,%,240,241,质谱图,每个点有一张唯一的质谱图,色谱图上的质谱图,数据观察-色谱图,样品运行中的每秒我们可获得一张或数张质谱图,通常是把检测到的离子叠加在一起获得一个唯一的总的数值,这些总的数值然后排列在一起,(,点对点,),产生一个类似的色谱图,(,TIC).,每次扫描加和所有的离子得到,.,总离子色谱图(,TIC),数据观察-色谱图,我们也能显示某一张质谱图,这让我们一个化合物的信息,质谱图,质量色谱图和,TIC,TIC,Mass 240,1.40,1.60,1.80,2.00,2.20,2.40,2.60,2.80,3.00,Time,0,100,%,0,100,%,240.27,1.41,e5,TIC,4.67,e5,消除背景的影响,增加信噪比,分离共流出物峰,数据观察-选择离子色谱图,TIC,图上显示的两个接近的流出峰，但每个峰是具有不同质谱指纹的不同化合物,.,数据观察-选择离子色谱图,11.00,12.00,Time,0,100,%,TIC,1.57,e5,数据观察-选择离子色谱图,100,200,300,400,500,600,m/z,0,100,%,254.2,194.2,507.3,334.3,399.7,100,200,300,400,500,600,m/z,0,100,%,194.2,580.4,402.2,226.2,460.2,11.00,12.00,Time,0,100,%,TIC,1.57,e5,分离共流出物峰,10.50,11.00,11.50,12.00,12.50,Time,0,100,%,0,100,%,0,100,%,254.23,5.81,e4,194.20,6.14,e4,TIC,1.57,e5,质量254,质量 194,TIC,选择离子色谱图,提取出离子给出更好的信噪比,消除背景影响,分离共流出物峰,提取出离子也能用于,:,峰的跟踪,准确定量,峰跟踪的实例,0,100,%,TIC,2.21,e6,1.00,2.00,3.00,4.00,5.00,6.00,Time,0.52,5.65,3.82,3.25,1.67,2.00,5.59,4.95,4.25,5.72,5.81,爆炸混合物的,TIC,哪一个是,HMX?,HMX,的质谱图,峰跟踪,40,60,80,100,120,140,160,180,200,220,240,260,m/z,0,100,%,84,72,59,69,113,89,106,237,131,205,158,分子离子峰,峰跟踪,质量色谱图,m/z 237,TIC,1.00,2.00,3.00,4.00,5.00,6.00,7.00,8.00,9.00,Time,0,100,%,0,100,%,2.00,0.52,5.65,3.82,3.25,1.67,2.00,4.95,4.25,5.81,分离相邻峰的能力,分子离子,M+H,+,M,.+,M-H,-,分辨率,FWHM,10%,峰谷,m/,M,M+H,+,m/z,intensity,质谱分辨率,复合技术,GC/MS,气相色谱-质谱,LC/MS,液相色谱-质谱,MS/MS,质谱-质谱,MS-MS,方法,MS 1,MS 2,固定扫描,子离子扫描,固定,母离子,扫描,扫描,中性丢失,固定,固定,选择离子监测,结果,MS,n,50,100,150,200,250,300,350,m/z,0,100,%,284,282,142,107,71,249,214,177,253,288,50,100,150,200,250,m/z,0,100,%,50,100,150,200,250,m/z,0,100,%,50,100,150,200,250,m/z,0,100,%,碎片,的碎片,的碎片,.,灵敏度,?,四极杆质谱仪如,TRACE MS,和,AQA,在全扫描模式下能分析到低于,pg,级的水平。,四极杆仪器最终的灵敏度在选择离子,(SIM),模式下能分析到低至,fg,级的水平。,什么是选择离子监测,-SIM?,代替全质谱范围的扫描，质谱仪允许仅在数个质量数范围扫描。,正如我们集中在数个质量数范围扫描，我们能够采集更长的时间。,与全扫描相比，,SIM,灵敏度可提高,10,到,100,倍。,这是四极杆质谱仪的特点,什么是,SIM,模式?,你不采集全质谱图,你仅采集样品中特征离子,分子离子,特征碎片,提高置信度,(,离子的比率,),仪器用更多的时间采集有用的数据使得灵敏度显著地改善。,采样频率快于全扫描，所以定量精度得到改善。,更少的样品更好的信噪比,全扫描和,SIM,8.300,8.400,8.500,8.600,8.700,8.800,8.900,9.000,9.100,time,0,100,%,0,100,%,2,pg,六氯苯，全扫描,选择离子,m/z 283.8,100,fg,六氯苯,SIM of m/z 283.8,SIM-,缺点,没有质谱图,-,不能用于未知物分析,你必须知道哪个离子需要采集,方法设置需要更多的时间,SIM,仅对目标化合物有用途,质谱仪的概要,质谱仪能给我们,.,分子量信息,A,指纹,帮助鉴定化合物,消除背景的影响,分离共流出物峰,示踪色谱峰,Shimadzu can provide customer withoptimized combination of LC and MS,excellent semi-micro,HPLC performance,rugged MS instrument,for routine work,