Ionization Methods in Mass Spectrometry at the SCS Mass Spectrometry Laboratory
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1 Ionization Methods in Mass Spectrometry at the SCS Mass Spectrometry Laboratory Steven L. Mullen, Ph.D. Associate Director
2 SCS Mass Spectrometry Laboratory Contact Information 31 oyes Laboratory (8:00-5:00 M-F)
3 SCS Mass Spectrometry Laboratory Staff Vera Mainz, Ph.D. (Acting Director) Steven L. Mullen, Ph.D. (Associate Director) Furong Sun Rachel Taylor Richard Milberg, Ph.D.
4 SCS Mass Spectrometry Laboratory Instrumentation Micromass 70-VSE (3) Micromass 70-SE-4F Micromass Quattro Micromass Q-Tof Ultima Applied Biosystems Voyager DE-STR Finnigan LCQ Deca XP
5 SCS Mass Spectrometry Laboratory 70-VSE (A) 70-VSE (B) 70-VSE (C) 70-SE-4F
6 SCS Mass Spectrometry Laboratory Quattro Q-Tof Voyager LCQ Deca
7 Information from Mass Spectrometry 1. Molecular Weight From molecular ion 2. Structure M + or (M+H) + [also (M+a) + or (M+K) + ] From fragments Hard ionization MS/MS 3. Elemental Composition From exact mass
8 Parts of a Mass Spectrometer Inlet Source Analyzer Detector Probe Leak Valve Syringe Plate GC LC EI CI FI FD FAB MALDI ESI Sector Quadrupole FT TF Ion Trap Electron Multiplier Photomultiplier Array Detector
9 Parts of a Mass Spectrometer Analyzer Source Detector Inlet
10 How a Mass Spectrometer Works Source Analyzer Detector
11 Interpreting a Mass Spectrum H 3 C CH 3 Acetone MW=
12 Isotopic Peaks 12C C6H6 13C 35Cl C6H5Cl 37Cl 79Br 81Br C6H5Br
13 Resolution Leu-enkephalin Monoisotopic Mass = Average Mass = R= R= R=100
14 Charge Leu-enkephalin Monoisotopic Mass = ion m/z = ion m/z =
15 HR Exact Mass Measurement H H CH 3 H H H C 21 H 22 4 ominal Mass: 338 Exact Mass: Found Mass: (Error = 0.1 ppm) C 22 H 26 3 ominal Mass: 338 Exact Mass: Found Mass: (Error = 1.0 ppm)
16 Ionization Techniques Short name Full name How it works EI Electron Ionization Impact of 70 ev electrons CI Chemical Ionization Transfer of H + + from CH 5 FI Field Ionization High Field (12kV) FAB Fast Atom Bombardment Beam of Xenon atoms FD Field Desorption High Field (12kV) MALDI Matrix-Assisted Laser Desorption Ionization Laser hits sample in UV-absorbing matrix ESI Electrospray Ionization Spray and desolvation of preformed ions
17 Ionization Technique Summary Ionization Volatile Thermal Size Amount Examples EI Yes Stable Small 1-2 mg rganics CI Yes Stable Small 1-2 mg rganics FI Yes Stable Small 1-2 mg rganics FAB o Labile Medium 50μg-1 mg FD o Labile Medium 1-2 mg Polar/ionic organics, peptides, biomolecules, organometallic on-polar organics, organometallics, porphyrins, PAHs MALDI o Labile Large 250fmol- 500pmol Peptides, proteins, DA/RA, polymers, dendrimers, fullerenes ESI o Labile Large 1-300pmol/μL Polar/ionic organics, peptides, proteins, biomolecules, organometallics, polymers
18 Electron Ionization (EI) Short name Full name How it works EI Electron Ionization Impact of 70 ev electrons Ionization Volatile Thermal Size Amount Examples EI Yes Stable Small 1-2 mg rganics M - e - M + Small stable organics Extensive fragmentation, 70eV libraries available HR available
19 EI Sample H Chemical Formula: C 11 H 12 2 Exact Mass: Molecular Weight:
20 EI Sample (Typical EI sample) Chemical Formula: C 12 H 12 2 Exact Mass: Molecular Weight:
21 EI Sample Chemical Formula: C 35 H Exact Mass: Molecular Weight:
22 EI Sample H Chemical Formula: C 24 H 28 2 Exact Mass: Molecular Weight:
23 EI Sample Br Chemical Formula: C 13 H 17 Br Exact Mass: Molecular Weight: (Typical EI sample)
24 Chemical Ionization (CI) Short name Full name How it works CI Chemical Ionization Transfer of H + from CH 5 + Ionization Volatile Thermal Size Amount Examples CI Yes Stable Small 1-2 mg rganics M + CH 5+ (M+H) + Small organics that fall apart in EI Extensive fragmentation HR available
25 CI Sample H Chemical Formula: C 10 H 14 4 Exact Mass: Molecular Weight:
26 CI Sample H Chemical Formula: C 18 H 25 3 Exact Mass: Molecular Weight:
27 CI Sample H Si(C 2 H 5 ) 3 Si(C 2 H 5 ) 3 Chemical Formula: C 18 H 38 4 Si 2 Exact Mass: Molecular Weight:
28 CI Sample H 2 H 2 Chemical Formula: C 19 H 34 2 Exact Mass: Molecular Weight:
29 CI Sample H (Typical CI sample) Chemical Formula: C 17 H 32 3 Exact Mass: Molecular Weight:
30 Field Ionization (FI) Short name Full name How it works FI Field Ionization High Field (12kV) Ionization Volatile Thermal Size Amount Examples FI Yes Stable Small 1-2 mg rganics M + kv M + Small organics that fall apart in EI, Isotopic enrichment Little fragmentation HR not available
31 Field Desorption (FD) Short name Full name How it works FD Field Desorption High Field (12kV) Ionization Volatile Thermal Size Amount Examples FD o Labile Medium 1-2 mg on-polar organics, organometallics, porphyrins, PAHs M + kv M + on-polar compounds o fragmentation HR not available
32 FD Sample H (Typical FD sample) Chemical Formula: C 48 H Exact Mass: Molecular Weight:
33 FD Sample Chemical Formula: C 37 H Exact Mass: Molecular Weight:
34 FD Sample Chemical Formula: C 12 H Exact Mass: Molecular Weight:
35 FD Sample (C) 4 s H (C) 3 s s(c) 3 H Chemical Formula: C 10 H 2 10 s 3 Exact Mass: Molecular Weight: (Typical FD sample)
36 FD Sample Chemical Formula: C 42 H 24 Exact Mass: Molecular Weight:
37 Fast-Atom Bombardment (FAB) Short name Full name How it works FAB Fast Atom Bombardment Beam of Xenon atoms Ionization Volatile Thermal Size Amount Examples FAB o Labile Medium 50μg-1 mg Polar/ionic organics, peptides, biomolecules, organometallic M + H + (M+H) + M + M + Polar/ionic compounds in liquid matrix (DTT/DTE or 3-nba) Some fragmentation HR available Rarely used, superseded by ESI
38 FAB Sample H H Chemical Formula: C 53 H Exact Mass: Molecular Weight:
39 Electrospray Ionization (ESI) Short name Full name How it works ESI Electrospray Ionization Spray and desolvation of preformed ions Ionization Volatile Thermal Size Amount Examples ESI o Labile Large 1-300pmol/μL Polar/ionic organics, peptides, proteins, biomolecules, organometallics, polymers M + H + (M+H) + M + M + M - H + (M-H) - M - M - Polar/ionic compounds, very little sample needed Little fragmentation HR available
40 ESI Sample H 2 H C H 2 C CH H 2 C H H H 2 C H 2 C H 3 C H 2 C CH CH 3 C H CH C H CH C H H 2 C CH C H CH C Angiotensin Asp-Arg-Val-Tyr-Ile-His-Pro-Phe-His-Leu Chemical Formula: C 62 H Exact Mass: Molecular Weight: H CH C C H H C CH H C CH H C CH CH 2 H 2 C H 2 C CH 3 CH CH 3 H CH CH 3 CH 2 H CH 2 CH 3 (Typical ESI sample) H
41 ESI Sample H Chemical Formula: C 21 H 32 7 Exact Mass: Molecular Weight:
42 ESI Sample 2 Chemical Formula: C 32 H Exact Mass: Molecular Weight:
43 ESI Sample H H Ir S PF 6 (Typical ESI sample) Chemical Formula: C 31 H 37 F 6 Ir 2 2 PS Exact Mass: Molecular Weight:
44 ESI Sample H Chemical Formula: C 12 H Exact Mass: Molecular Weight:
45 Matrix-Assisted Laser Desorption Ionization (MALDI) Short name Full name How it works MALDI Matrix-Assisted Laser Desorption Ionization Laser hits sample in UV-absorbing matrix Ionization Volatile Thermal Size Amount Examples MALDI o Labile Large 250fmol- 500pmol Peptides, proteins, DA/RA, polymers, dendrimers, fullerenes M + laser (M+H) + Large compounds, very little sample needed Little fragmentation HR not available
46 MALDI Sample Tg Tg Tg Tg Tg MW=1907 Tg
47 MALDI Sample C 2 Tg TIPS 12 TIPS MW=3822
48 MALDI Sample H 2 H H H H H H P - - n MW=34,000
49 MALDI Sample H 2 -U PS TFPGHSALMICGTLTLK-H MW=1852
50 MALDI Sample Br H = * H H * 7 MW = 1270
51 MS Experiment Summary Experiment Ionization Comments LR All Mass Spectrum HR EI, CI, FAB, ESI Exact Mass Elemental Composition MS/MS ESI, FAB, MALDI Structure, e.g. Peptide Sequence Isotope Ratio FI Isotopic Enrichment GC/MS EI, CI, FI LC/MS ESI HPLC or Capillary HPLC
52 Graphic by Dorothy Loudermilk
53 Further Information Mass Spectrometry: A Textbook Jürgen H. Gross 2004, Springer-Verlag, ISB Interpretation of Mass Spectra, 4 th edition F. W. McLafferty 1993, University Science Books, ISB Mass Spectrometry for Biotechnology Gary Siuzdak 1996, Academic Press, ISB
54 Use of this file This slide show was prepared by Steven L. Mullen, Ph.D. Mass Spectrometry Laboratory School of Chemical Sciences University of Illinois (217) It may be used for non-commercial educational purposes, with appropriate credit and no alterations.
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