1.串联液质系统分析原理

串联液质系统分析原理美国应用生物系统中国公司 应用技术服务部Opportunities in the Services and System Solutions MarketWhat is a mass spectrometer ?2Date | AB CONFIDENTIAL© 2006 Applied Biosystems1Opportunities in the Services and System Solutions Market质谱技术：● 质谱分析过程：样品通过进样系统进入离子源，由于结构 性质不同而电离为各种不同质荷比（m/z）的分子离子和碎 片离子，而后，带有样品信息的离子碎片被加速进入质量 分析器，不同的离子在质量分析器中被分离并按质荷比大 小依次抵达检测器，经纪录即得到按不同质荷比排列的离 子质量谱，也就是质谱 (mass spectrum)。

● 质谱：按离子的质荷比的大小依次排列形成的图谱3Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market质谱仪的主要部件： 真空系统 数据系统样品导入 系统直接进样 GC LC CE离子源质量分析器离子检测器MALDI ESI API APCI APPI四极杆：Quadrupole 离子阱、线性离子阱：IT，LIT 飞行时间：TOF 傅立叶变换离子回旋共振：FT-ICR4Date | AB CONFIDENTIAL© 2006 Applied Biosystems2Opportunities in the Services and System Solutions Market高效分离系统高灵敏度检测系统GC 大量的载气EI、CI…MS 高真空体系HPLC 大量的液态流动相？MS/MS 高真空体系5Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market液相色谱与质谱的联用技术● 主要障碍：1、能否把大量的溶剂除去并不使质谱仪受到污染 2、能否使被分析样品在温和条件下带上电荷得到分析而 不被分解上面两点是影响灵敏度的主要因素GS2 TEMGS1 GS2 TEM1、GS2↑，TEM↓ 2、H2O比例大时，TEM ↑，有利于离子化6Date | AB CONFIDENTIAL© 2006 Applied Biosystems3Opportunities in the Services and System Solutions MarketTriple Quadrupole sequence of eventsIon production Ion transport Ion filtering Ion processing Ion filtering Ion detection9Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market1st Step：Ion ProductionIon production10Date | AB CONFIDENTIAL© 2006 Applied Biosystems4Opportunities in the Services and System Solutions MarketPrinciple of Electrospray (ESI)IonSpray™ source: Electrospray Ionization (ESI) softest ionization technique polar to ionic substances11 Date | AB CONFIDENTIAL © 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketPrinciple of Atmospheric Pressure Chemical Ionization (APCI)Heated Nebulizer source: Atmospheric Pressure Chemical Ionization (APCI) corona discharge needle polar to unpolar and thermally stable compounds12 Date | AB CONFIDENTIAL © 2006 Applied Biosystems5Opportunities in the Services and System Solutions Market2nd Step：Ion transportIon transport13Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketCurtain Gas™ interfaceATMOSPHERIC PRESSURECUR DP SK Q0 + + FP + + + + + + +VACUUM+++ + + +++++++++N21. Assists in droplet evaporation 2. Declusters ions – CID (in-source) fragmentation 3. Collisional focussing in Q014 Date | AB CONFIDENTIAL © 2006 Applied Biosystems6Opportunities in the Services and System Solutions Market抗污染 LC流速大时，CUR↑ 助气化DP： 不同化合物，DP不同，应分别优化 DP设置过大，化合物会被部分打碎——源内裂解15Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market3rd Step：Ion FiletingIon filtering16Date | AB CONFIDENTIAL© 2006 Applied Biosystems7Opportunities in the Services and System Solutions MarketHow are ions moved in a quadrupole ?Radio frequency (RF) Alternating current / direct current (AC/DC) Ion energy17Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market– ++ +++ + ++ –1. ... A Movement of allows only one m/z 6. RF-voltage changes into system to pass 5. Electrical DCthe ioninto direction of the 4. Ion of AC of quadrupoledirection respectively, 3. Movement/thethe ion polarity and electrical repulsion 2. pair entersrepulsion and attraction, of the nearest the quadrupole rod with rodsopposite charge charge and attraction, All other m/z areion nearest quadrupole rod with between quadrupole between quadrupole the and destabilized. quadrupole. respectively, the opposite rods and ion18 Date | AB CONFIDENTIAL © 2006 Applied Biosystems+–– –+++– – + + – + + – – +8– –+ +Opportunities in the Services and System Solutions MarketHow are ions moved in the quadrupole ?Radio frequency (RF) Alternating current / direct current (AC/DC) Ion energy19Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketIon energy spirals ions through the quadrupole.20 Date | AB CONFIDENTIAL © 2006 Applied Biosystems+ + + + ++ + + + + ++ + + + ++ + + + ++ + + + ++ + + + + ++ + + + ++ + + + ++ + +9Opportunities in the Services and System Solutions MarketMS: modes of operationQ1 full scanscan Precursor IonsSelected Ion Monitoring (SIM)select Precursor Ion21 Date | AB CONFIDENTIAL © 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketQuadrupole：full scanning mode1st scanLast m/z2nd scanLast m/zm/z ratiom/z 3 m/z 2 m/z 1m/z 3 m/z 2Time22 Date | AB CONFIDENTIAL © 2006 Applied Biosystems10Opportunities in the Services and System Solutions MarketQuadrupole: filtering mode1st experimentm/z 32nd experimentm/z 3m/z ratio• Higher dwell time per m/z • Higher selectivity • Higher S/Nm/z 2 m/z 2m/z 1m/z 1Time23 Date | AB CONFIDENTIAL © 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market扫描时间的含义： 1、仪器扫描频率一定 2、扫描时间，指采集图谱的时间间隔，即该时间内 多个扫描循环获得的所有图谱进行平均，并记录该 平均图谱 扫描时间的设定： 1、全扫描： Parameter range，根据质量范围大小，设为1～2sec Center/Width，6～10Da/amu，设为0.5sec 2、MRM： ～100msec/transition，可根据离子对灵敏度调整24Date | AB CONFIDENTIAL© 2006 Applied Biosystems11Opportunities in the Services and System Solutions Market4th Step：Ion ProcessingIon processing25Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketMS/MS: LINAC® collision cell● Precursor ion flies into collision cell ● Collides with collision gas (N2) ● Fragments into a charged and a neutral fragment ● Product ions will be detected in Q3entranceexit• Linear Accelerator: Electrical field gradient will accelerate product ion after collision26 Date | AB CONFIDENTIAL © 2006 Applied Biosystems12Opportunities in the Services and System Solutions MarketPower of LINAC® collision cell100 pesticides in fruit and vegetable Calibration 0.001 – 0.1mg/kg 25msec dwell time per MRM27Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions Market5th Step：Ion FilteringIon filtering28Date | AB CONFIDENTIAL© 2006 Applied Biosystems13Opportunities in the Services and System Solutions MarketMS/MS: modes of operationProduct Ion Scan Precursor Ion Scanfilter Precursor Ion scan Product Ionscan Precursor Ion filter Product IonNeutral Loss ScanMultiple Reaction Monitoring (MRM)29Date | AB CONFIDENTIALscan Q1 and Q3 with constant mass off-set© 2006 Applied Biosystemsfilter Precursor Ion filter Product IonOpportunities in the Services and System Solutions Market6th Step：Ion DetectionIon detection30Date | AB CONFIDENTIAL© 2006 Applied Biosystems14Opportunities in the Services and System Solutions MarketCEM Gain Check (Q1 SIM)31Date | AB CONFIDENTIAL© 2006 Applied BiosystemsOpportunities in the Services and System Solutions MarketThanks for listening !32Date | AB CONFIDENTIAL© 2006 Applied Biosystems15。