离子飞行的平均自由程需最大化.pdf

1 WW Seminar Tour Spring 2006 Liquid Chromatography Mass Spectrometry Fundamental Jimmy Chan WW Seminar Tour Spring 2006 A general detector It can connect with HPLC to separate and get more structure informations It can use in quantitative and qualitative analysis The Advantage of LC MS PDF created with pdfFactory trial version 2 WW Seminar Tour Spring 2006 The Limit of LC MS Sample must be ionization Ionic compound can motion transfer and analysis MS must vacuum 离子飞行的平均自由程需最大化离子飞行的平均自由程需最大化 理论上 真空度越高越好理论上 真空度越高越好 MS must analysis with M Z WW Seminar Tour Spring 2006 Suitable for both ES and APCI Methanol Acetonitrile WaterWater Ethanol Propanol Isopropanol Butanol DMF 1 DMSO 1 Acetone CH2Cl2 CHCl3 Suitable for only APCI Toluene Benzene 2 Hydrocarbons e g Hexane Styrene 2 CCl4 CS2 Cyclic Hydrocarbons e g Cyclohexane 1 At lower solvent percentages 10 or less under ES conditions 2 These solvents not often used due to environmental health and safety issues The Solvent of API MS PDF created with pdfFactory trial version 3 WW Seminar Tour Spring 2006 The Selection of APIThe Selection of API AddtiveAddtive pH 乙酸 甲酸 TFA 氢氧化铵 一般的缓冲液一般的缓冲液 离子配对试剂离子配对试剂 乙酸铵 甲酸铵 三氟乙酸 TFA 七氟丁酸 HFBA 四乙基或四丁基氢氧化铵 TBAH 阳离子化试剂阳离子化试剂 20 50 M的乙酸钠或乙酸钾 一般考虑一般考虑 挥发性 污染喷雾室 堵塞喷嘴 导电性 对离子气化过程减少小液滴的形成 离子配对 进入气相时中和预带电离子 不能使用在不能使用在不能使用在不能使用在HPLCHPLC中常使用的磷酸盐 硫酸盐或硼酸盐中常使用的磷酸盐 硫酸盐或硼酸盐中常使用的磷酸盐 硫酸盐或硼酸盐中常使用的磷酸盐 硫酸盐或硼酸盐 WW Seminar Tour Spring 2006 The Application of wide range Proteomics Peptide and Protein molecular Weight Peptide Sequence Peptide and Protein Purity Analysis Protein identification Small Molecular Medical drug Analysis Drug abuse Analysis Antibacterial drug Analysis Steroid Analysis Structure comformation PDF created with pdfFactory trial version 4 WW Seminar Tour Spring 2006 非挥发性的缓冲盐非挥发性的缓冲盐挥发性缓冲盐挥发性缓冲盐 浓度浓度 10 mM 对于对于 ES 或或 溶剂溶剂 H 溶剂溶剂 H M 溶溶 剂剂 M WW Seminar Tour Spring 2006 The Characteristic Of API ESI Compound with volitility Compound with thermo unstabilite Ionization form in liquid Besides singal charge it can form mutiple charges APCI Compound with volitility Compound polarity lower than ESI Compound with thermo stabilite Ionization form in gas Only singal charge PDF created with pdfFactory trial version 6 WW Seminar Tour Spring 2006 The Relative Application of LC MS WW Seminar Tour Spring 2006 What is LC MS PDF created with pdfFactory trial version 7 WW Seminar Tour Spring 2006 MW 308 100 120 140160 180 200 220240 260 280 300 320 340 360 380 400 420 6000 14000 22000 30000 38000 46000 m z Abundance 309 M H 苯基丁氮酮 Phenylbutazone 的API ES 谱图 Which mode do you get the singal charge 基峰 谱图中最强的离子 M H WW Seminar Tour Spring 2006 Ginsenoside Which mode do you get the singal charge PDF created with pdfFactory trial version 8 WW Seminar Tour Spring 2006 6507007508008509009501000105011001150 12001250 Abundance 5000 10000 15000 20000 25000 30000 35000 40000 m z A 25 679 45 A 24 707 65 A 23 738 45 A 22 771 90 A 21 808 60A 20 848 95A 19 893 55A 18 943 15A 17 998 55 A 16 1060 85 A 15 1131 50A 14 1212 30 Abundance 16400166001680017000172001740017600m z 40000 80000 120000 160000 200000 240000 280000 A 16959 09 Assume adduct is a proton M1 A n n M2 A n 1 n 1 Solve for A Which mode do you get the mutiple charge 肌红蛋白 Myoglobin 的API ES 谱图 WW Seminar Tour Spring 2006 Mass Spectrum Concept COOH OH CF3 COOH OCOCH3 CF3 反应产物的联机分析结果 a 色谱图 b 总离子流图 d 质谱棒图 Time m z Abundance 3D Data R T m z Intensity TIC PDF created with pdfFactory trial version 9 WW Seminar Tour Spring 2006 Raw Filtered Full and Condensed Data Raw Data 原始数据原始数据 Filtered Full Data 过滤后的数据过滤后的数据 Profile Condensed Data 精简数据精简数据 line spectra Abund Abund Abund m z m z m z threshold WW Seminar Tour Spring 2006 Mass Spectrum PDF created with pdfFactory trial version 10 WW Seminar Tour Spring 2006 Total Ion Chromatography TIC UV TIC WW Seminar Tour Spring 2006 LCDEMO 02 D EIC 285 ll MS Smoothed 2 4 3 SG LCDEMO02 D BPC 100 400 ll MS Smoothed 2 4 3 SG 0 00 0 25 0 50 0 75 1 00 1 25 6 x10 Intens 0 0 0 5 1 0 1 5 2 0 2 5 3 0 7 x10 123456T ime min Increase S N Subtract background Separate the co elution What the purpose of EIC Quantitative Extract Ion Chromatography EIC EIC 285 TIC PDF created with pdfFactory trial version 11 WW Seminar Tour Spring 2006 The Explain of API Mass Spectrum ESI and APCI can supply the compound messgae The ion depend on additive and condition Positive M H 酸性条件酸性条件 M Na M K 有盐时有盐时 M NH4 有铵盐缓冲溶液有铵盐缓冲溶液 M X X 溶剂或缓冲溶液中的阳离子溶剂或缓冲溶液中的阳离子 2M H 在高浓度时形成的二聚体在高浓度时形成的二聚体 M H S 溶剂添加剂溶剂添加剂 Negative M H 碱性条件碱性条件 M X X 溶剂或缓冲溶液中的阴离子溶剂或缓冲溶液中的阴离子 M H S 溶剂添加剂溶剂添加剂 WW Seminar Tour Spring 2006 What is the Collision Induced Dissonciation CID 通过与中性分子碰通过与中性分子碰撞将撞将能能量传递给量传递给离子的过离子的过程 程 能能量量 传递足传递足以以导致键导致键的的开开裂裂和所选和所选离子的离子的重排 重排 PDF created with pdfFactory trial version 12 WW Seminar Tour Spring 2006 ION TRAP The CID in API Transition region 这是什么这是什么 利用 API MS 的传输区的电场在中压区 0 01 1 torr 碰撞激活离子来 产生CID质谱图 硬件硬件 WW Seminar Tour Spring 2006 The Process of HED and EM 正正离子离子采样采样 负负离子离子采样采样 Lens Dynode 7 kV EM 0 to 3000 EM 0 to 3000 Lens e e Dynode 7 kV SQ SQ PDF created with pdfFactory trial version 13 WW Seminar Tour Spring 2006 Introducing the Agilent 6130 Singal Quadrupole Mass Spectrometer WW Seminar Tour Spring 2006 6300 Series Ion Trap LC MS 6210 Time of Flight LC MS 6510 Quadrupole Time of Flight LC MS New QQQ New QTOF 6410 Triple Quadrupole LC MS New Agilent 6000 LC MS Series A new and expanded portfolio for applications from life science to chemical analysis combining exceptional performance and legendary Agilent reliability 6100 Series Single Quad LC MS New Single Quads Updated Ion Trap Family Updated TOF PDF created with pdfFactory trial version 14 WW Seminar Tour Spring 2006 Lassen LC MS Singal Quadrupole system introduction WW Seminar Tour Spring 2006 Atmosphere Pressure Ionization Mass Spectrometry API MS API MS is usde for analysis liquid samples that introduce by HPLC FIA and Infusion Application in wide mass range 1pg sensitiveity limit PDF created with pdfFactory trial version 15 WW Seminar Tour Spring 2006 6100 Series SQ 6110 6120 6130 and 6140 1 Entry Level 10 to 1500amu p n G6110AA 2 Mid range POS NEG 10 to 1500amu p n G6120AA 3 Research 2 to 3000amu p n G6130AA 4 Research fast scan 10 to 1350amu p n G6140AA 6120 SIM Scan 10pg S N 20 1 Mass range 10 1500 m z Pos Neg Switching and Muti Signal Acquisition WW Seminar Tour Spring 2006 Function Block Diagram Inlets Ion Source Ion OpticsMass Filter Detecto r Vacuum System Electronics PDF created with pdfFactory trial version 16 WW Seminar Tour Spring 2006 Agilent LC 6320 Ion Trap MS Neutral Molecules Analyte Ions Clusters Salts capillary Liquid introduce Nubalizer Heat N2 Nubalizer gas 碎片碰撞电碎片碰撞电 压压 CID Shield hole Octopole Quadrupole HED EM Focus Len WW Seminar Tour Spring 2006 Mutimod Source ESI APCI Capillary HPLC inlet Nebulizer Drying gas Corona needle ESI Zone APCI Zone Thermal container IR emitters Charging electrode Reversing electrode Sensor for vapor temperature APCI counterelectrode PDF created with pdfFactory trial version 17 WW Seminar Tour Spring 2006 The G1978A Multimode Source APCI corona needle Nebulizer Field shaping electrodes IR emitters Sensor for vapor temperature APCI Counterelectrode WW Seminar Tour