LC MSMS based MSMS market survey MSMS Terminology
LC MSMS based 주요 기능 및 분석 요약 MSMS market survey용
내용 * MSMS Terminology 1. MS의 주요 Data, 기능 및 Spec. * 농약시료의 분석특징 2. 시료 전처리 및 분취의 필요성 3. MS, MSn 의 응용 * Conventional LC vs UPLC 3. 1 MRM mode 3. 2 Product Ion Scan(SIM/SCAN) 3. 3 Information/Data Dependant Analysis 3. 4 MS 3 의 활용 3. 5 Precusor Ion Scan (SCAN/SIM), Neutral Loss (SCAN/SCAN) 3. 6 Mass Defect Filtering 4. MSMS의 주요 응용 영역 MSMS market survey용
MSMS Terminology Sample 주입 Ion Source Ion Guide Orthogonal In-source CID GC LC CE ICP DIP EI/CI ESI/APCI/APPI MALDI Odd Ion Even Ion Cluster ion m/z Target 정제 분취 Signal Data Spectrum Solution Extraction 시료 전처리 Column , … PDA MS MSMS , … Focus Accelation Ion Mobiility MS 1 Separation 1 _ Separation/Purification Cell MS 2 Frag. Separation 2 CID Ar N 2 SQ TOF Trap(3 D) LIT(2 D) Triwave Ion Mobility Multiplier SQ TOF Trap(3 D) LIT(2 D) Detection EM PM MCP Off-axis Orbitrap Accurate Mass : 1 -수백 ppm, SIM Chromatogram/SCAN Spectrum, Formula, Structural information, Isotope Ratio, … Mass Resolution : FWHM, HR/LR Identification Point System m/z Separation : gas phase 분자이온의 경로/속도차 분석시간 : 수 ms- 수 십 ms 정성(m/z)/정량 감도 Sub PPB (LOQ), LOD 진공 10 -5 -10 -7 torr MSMS market survey용 Data System Qual/Quan Matrix MC Isotope LC Library/DB Deconvolution Profiling s/w PCA
1. MS의 주요 Data, 기능 및 Spec. Mass Filter SQ TOF SIM chromatogram Yes No XIC Yes 정량 MS Scan spectrum Yes(Resolution) Yes(Trap) 정성 MSn No In-source CID Post source decay Yes 정성 Mass Resolution 1 Da 수 mili-Dalton 0. 1 Da 정성 Mass Accuracy 수백 ppm 수십ppm 정성 주요기능 Ion Trap(3 D) 정성/정량 • MSn : MRM(SIM/SIM)/Scheduled MRM, SRM, MS 3, Product Ion Scan(SIM/SCAN), Precusor Ion Scan(SCAN/SIM), Neutral Loss(SCAN/SCAN), Information/Data dependant analysis MSMS market survey용
Enhanced MS Scan _ Deconvolution M+Z+2 Ab M+Z+3 Calculation Equation M+Z+1 M+Z+4 M+Z MW 2 MW 1 = (M+Z) / Z MW 2 = (M+(Z+1)) / Z+1. . M = 26. 83 K Da MW 1 MS Scan sensitivity Pure Protein from Prep LC ? MSMS market survey용
2. Sample Matrix Effects in Pesticide Analysis Targets in Sample Matrix : solid/liquid/gas polar/non-polar/ionic/small/large compounds 추출 Clean-up 용매 추출 회수율 방해요인 1. 고체 덩어리 2. 추출 용매에 녹지 않는 성분 3. 온도 변화, Chemical Reaction, … 정성 정량 선택적인 검출/정량 방해요인 1. Signal Addition : Ghost peak, RT shift, => Matrix Matched Calibration 2. Signal Suppression : Ion Suppression, … => Isotope Labeled Standard Calibration MSMS market survey용
Reducing and Changing of Sample Matrix(Interference) 1. Universal Extraction : 다 성분 분석, 농약 원재료 잔류물 분석등 Sample Matrix 용매추출 Interference 석출법 Chemical Dispersive Solid Phase Freeze Drying 재 용해, 필터 후 분석 2. Partial Extraction : 부분 성분 분석, 농약 Metabolites 잔류물 분석등 Sample Matrix 용매추출 Interference 제거 SPE흡착용 용매로 희석 On-line SPE? SPE 흡착 선택적 용리 필터 후 분석 선택적 3. Selective Extraction : Impurity 분취, Prep Process(Prep LC, HPCPC, …) , Im. AC(Mycotoxin, …), MIP, … MSMS market survey용
시료전처리 종류 및 분석장비 추출 Clean-up 정성 정량 Sohxlet Precipitation Ultrafiltration LLE LC MSMS GC MSMS (On/Off–line, Micro) Qu. ECh. ERS GPC/RAM SPE (On/Off–line, Micro) Multi-Residue Selectivity Clean-up & Concentration Matrix Effect Specificity Metabolites Immuno. AC MIP MSMS market survey용 (Compound Specific Information) LC MS GC MS (RT, 유도체화) LC FLD LC UV/Vis GC ECD/NPD GC FID
3. MSn to MSMS, Datadase구축? A. MSMS in Time : 농축(Trap/Enhanced)/Mass필터 - Ion Trap(3 D) : 1/3 Cut-off Ion Trap(3 D)-TOF Q-Trap (Hybrid Linear Ion Trap) LTQ-Orbitrap (Hybrid Linear Ion Trap) B. MSMS in Space - QQQ - Q-TOF - TOF-TOF MSMS market survey용
Conventional LC vs UPLC Porous Beads Totally Porous, : Column 충전물 Non-Porous Beads Superficially Porous, Surface Porous 11
Conventional LC vs UPLC : Van Deemter Plot Optimal Linear Velocity Column Downscale 12
Target Quan/Qual 3. 1 MRM mode Agilent QQQ 추가적인 정성 MSMS market survey용
많은 시료, 다 성분 screening : UPLC+s. MRMmode High Throughput ? AB Sciex MSMS market survey용
3. 2 Product Ion Scan Metabolic route에 따른 Mass Shift Phase I Metabolism Mass Shift Group Reaction P 32 O 2 Oxidation at C, N, S P 16 O P 14 O-H 2 Double bond N 2 -H 2 N 14 -CH 2 dealkylation N 28 -C 2 H 4 N 30 -O 2+H 2 Phase II Metabolism Mass Shift Group Reaction P 305 Glutathion Conjugation P 192 Glu. A Glucuronidation P 176 Glu. A Glucuronidation P 80 SO 3 Sulfation P 42 CH 2 CO Acethylation -H 2 O -C 4 H 4 Double Bond Oxidation Ring Opening MSMS market survey용 (SIM/SCAN) MSMS Scan Sensitivity Database ?
Flonicamid and metabolite of 파프리카
3. 3 Information/Data Dependant Analysis Thermo Finnigan LTQ AB Sciex Q-Trap MSMS 주요기능 Q 1 Q 2 Q 3 e. PI Scan SIM Fragment Trap, Scan MS 3 SIM Fragment Trap, Isolate, Fragment, Scan SRM SIM Fragment Scan MSMS market survey용
Information/Data Dependant Analysis MS SCAN and MSMS SCAN switching Inclusion/Exclusion list 764 MS-MS SCAN Survey SCAN Database 비교? 764 MSMS market survey용
Information/Data Dependant Analysis MS and MSn switching, Inclusion/Exclusion list MS Scan(Baseline noise) Exclusion : 402. 3, 418. 4 MS Scan(Target+Baselin) Inclusion : 262. 1, 358. 9, 376. 2 New metabolite MW 376 in Buspirone And Target ID MSMS market survey용
Information/Data Dependant Analysis SRM and MSMS SCAN switching Quantitation and Identification/Confirmation SRM 262(Buferalol) and 278(M 1, M 2, M 3, …) SRM Triggered Enhanced PI MSMS market survey용
Precusor Ion Scan/ e. PI Scan(remikiren-OH) (A) precursor ion scan trace of m/z 254 (B) enhanced product ion trace (A) Precursor ion spectrum of the peak at 3. 2 min (B) Enhanced product ion spectrum of the precursor ion at m/z 647 JOURNAL OF MASS SPECTROMETRY J. Mass Spectrom. 2004; 39: 845– 855 Published online in. Wiley Inter. Science (www. interscience. wiley. com). DOI: 10. 1002/jms. 659 MSMS market survey용
3. 4 MS 3 의 활용 The ion in the MS 3 spectrum at m/z 274. 1549 suggests GSH addition to the 2 -chloro-4 -piperazin-1 -ylphenol moiety. MSMS market survey용
3. 5 Precusor Ion Scan MS 1 CID MS 2 Tramadol m/z 264 => m/z 58 MSMS market survey용 (SCAN/SIM)
Neutral Loss MS 1 CID (SCAN/SCAN) MS 2 Scan TIC Neutral Loss MSMS market survey용 Neutral Loss 176 Da Glucuronic acid 80 Da Sulfate conjugate 132 Da , …
3. 6 Mass Defect Filtering Waters Q-TOF Mass Resolution/Accuracy? Metabolic route에 따른 Mass Shift Phase I Metabolism Phase II Metabolism Mass Shift Group Mass Defect Shift P 32 O 2 -0. 0102 P 305 Glutathion 0. 0682 P 16 O -0. 0051 P 192 Glu. A 0. 027 P 14 O-H 2 -0. 0106 P 176 Glu. A 0. 0321 N 2 -H 2 -0. 0157 P 80 SO 3 -0. 0432 N 14 -CH 2 -0. 0157 P 42 CH 2 CO 0. 0106 N 28 -C 2 H 4 -0. 0313 N 30 -O 2+H 2 0. 0055 0. 04 Da 이하 0. 07 Da 이하 MSMS market survey용
Mass Defect Filtering TIC Unfiltered Spectrum XIC Signal Purification MDF spectrum Mass Defect PCA, Profiling, … MSn with mass defect filtering for in vitro and in vivo metabolite identification MSMS market survey용
4. MSMS의 주요 응용 영역 1. Protein/Peptide 2. Metabolite 3. Natural Product Profiling 4. Unknown Analysis : Impurity 분석 5. Target Analysis : 유해 Regulation성분 6. etc, … MSMS market survey용
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