SCS Mass Spectrometry Laboratory Contact Information Staff 31 Noyes Laboratory (8:00-5:00 M-F) 217-333-2545 http://scs.illinois.edu/massspec/ Furong Sun (frs@illinois.edu) Furong Sun Director Training and LCMS, EI and CI Dr. Haijun Yao Spectrometrist ESI, FD/FI/LIFDI
Instrumentation and ionization techniques Submitted Samples Waters 70-VSE Double focusing Sector (3) (EI/CI, FAB) Water GCT Premier (EI, CI, FD, FI, LIFDI and GCMS) Waters Q-TOF Ultima (HR ESI, LCMS) Waters Synapt Q-TOF (HR ESI, LCMS, AP-GC) Thermo LTQ Orbitrap XL (ESI, DESI) Self-Service or Submitted Samples Waters Quattro Ultima Triple Quadrupole (LR ESI, LCMS, MRM) Waters ZMD Single Quadrupole (LR ESI) AB Sciex Voyager DE-STR TOF (MALDI) Bruker Daltonics UltrafleXtreme TOF/TOF (MALDI, TLC, MS imaging)
We offer the following services Low and High resolution ESI, FD/FI and EI/CI LR APCI, FD/FI, MALDI and LIFDI TLC MALDI MALDI imaging GC/MS LC/MS and LCMS/MS (MRM, DDA)
SCS Mass Spectrometry Laboratory 70-VSE Waters GCT These instruments are used for EI/CI, FD/FI, FAB and GC/MS.
SCS Mass Spectrometry Laboratory Quattro Ultima ESI/APCI Q-Tof - HR ESI, LCMS Orbitrap ESI, DESI ZMD - LR ESI Synapt HR ESI, APGC These instruments are used for LR and HR ESI and LCMS.
SCS Mass Spectrometry Laboratory Bruker Daltonics Autoflex Speed MALDI Bruker Daltonics UltrafleXtreme MALDI TOF-TOF
Mass Spectrometry for small molecules Molecular Weight From molecular ion M + or (M+H) + [also (M+Na) + or (M+K) + ] Structure From fragments Hard ionization MS/MS Elemental Composition From exact mass
Mass Spectrometry for biomolecules Molecular weight determination, structure information For example - Protein, peptides, DNA/RNA, Carbohydrates, Oligonucleotides, Oligosaccharide, Lipids, Natural products, Dendrimers, Porphyrins, Metalloporphyrins Identification, sequence and modifications For example - Protein, peptides, DNA/RNA, Carbohydrates, Oligonucleotides, Oligosaccharide, Lipids, Natural products
Parts of a Mass Spectrometer Inlet Source Analyzer Detector Probe Leak Valve Syringe Plate GC LC EI CI FI FD MALDI ESI APCI APPI Sector Quadrupole TOF Ion trap Orbitrap Hybrid Electron Multiplier Photomultiplier Array Detector MCP
Features of Electrospray MS Soft ionization ions are produced in solution It can be used for small and large molecules Multiple charged ions are usually produced for large molecules Suitable for using as an interface with HPLC Fast data acquisition High sensitivity, superior mass resolution and accuracy Sample must be soluble and stable in solution, polar relative clean
Features of MALDI-TOF MS Soft ionization - analyze intact biomolecules and synthetic polymers Broad mass range - analyze a wide variety of biomolecules Simple mixtures are okay Relatively tolerant of buffers and salts Fast data acquisition Easy to use and maintain, no water or gas hook ups required High sensitivity, superior mass resolution and accuracy
Ionization Technique Summary Ionization Volatile Thermal Size of molecules Amount Examples EI Yes Stable Small 1-2 mg Organics CI Yes Stable Small 1-2 mg Organics FI Yes Stable Small 1-2 mg Organics FD No Labile Medium 1-2 mg Non-polar organics, organometallics, porphyrins, PAHs MALDI No Labile Large 250fmol- 500pmol Peptides, proteins, DNA/RNA, polymers, dendrimers, fullerenes ESI No Labile Large 1-300pmol/mL Polar/ionic organics, peptides, proteins, biomolecules, organometallics, polymers
Flowchart of Mass Spectrometry Techniques To print - use the landscape setting.
Chromatogram LCMS Mass Spectra
LC Tandem Mass Spectrometry (LCMSMS) Chromatogram Mass Spectra
LCMS Data Dependent Analysis (DDA) NAS_01 Qtof_27783_dda 100 0% 0% 0% 0% Chromatogram 21-Jan-2010 21:30:55 28.46 39.66 49.89 2:1,B 4: TOF MSMS ES+ BPI 2.35e3 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 Qtof_27783_dda 3: TOF MSMS ES+ 100 56.84 65.60 BPI 2.03e3 28.98 37.97 47.61 49.85 53.48 61.96 63.88 70.23 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 Qtof_27783_dda 2: TOF MSMS ES+ 65.79 100 BPI 56.80 2.91e3 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 Qtof_27783_dda 1: TOF MS ES+ 65.75 100 BPI 56.73 6.96e3 28.86 38.16 53.37 44.21 46.34 48.7650.52 55.32 60.75 63.36 68.72 Time 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00 70.00 53.05 56.88 58.31 63.16 28.58 38.23 53.44 49.81 55.37 62.28 68.57 73.05 65.67 MS and MS2 NAS_01 Qtof_27783_dda 203 (53.626) 100 0% 0% 0% QTOF 4: TOF MSMS 679.53ES+ 54 Qtof_27783_dda 711 (53.366) 1: TOF MS ES+ 100 518.0 776.4 2.55e3 512.3 519.1 778.0 679.5 744.9 883.4 992.8 1488.7 1551.9 m/z 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 0% 322.3 435.3 548.4 Qtof_27783_dda 212 (53.786) 512.3 100 538.8 506.6 276.2 360.2 431.3 679.6 680.6 638.9 768.4 3: TOF MSMS 517.96ES+ 454 Qtof_27783_dda 310 (53.441) 2: TOF MSMS 776.43ES+ 625.4 100 336 626.4 767.9 276.1 360.2 686.4 459.2 778.0849.5 1076.6 573.3 910.5 962.6 1175.7 1276.7 1308.7