Mass Spectrometry - Faculté de médecine de l'université d'Ottawa

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2,5-Dihydroxy benzoic acid (DHB) Trans-3,5-dimethoxy-4-hydroxy cinnamic acid (commonly known as sinapic acid or Sinapinic acid, SA) 4-Hydroxy-α-cyanocinnamic acid Glycerol* * - common matrix for IR MALDI experiments 2,4,6-Trihydroxy acetophenone (THAP) Common chromophoric matrices used in UV MALDI with the noted exception of glycerol (in grey box), which is commonly used as a chromophoric matrix in IR MALDI. 11/21/2005 30
Sample plate 2-3 kV + + + + + + + + + + + + + + + + + + + + + ++ + + + N 2 laser (337 nm) + + + Grid or lens at lower voltage than plate + + + ++ + Video camera The matrix-assisted laser desorption ionization process. Laser ablation causes ions and neutrals to be desorbed and efficient ionization to occur. Ions are released into the gas-phase and their m/z ratios analyzed 11/21/2005 31 by mass spectrometry. Ions Ions Ions Ions Ions + + Neutral matrix + 1 + matrix 1 + matrix dimer + + + + ++ + + + + + Neutral analyte 1 + analyte + 1 + analyte dimer ++ 2+ analyte
Page 1 and 2: Mass Spectrometry D. Figeys The Ott
Page 3 and 4: A mass spectrum: a plot of the rela
Page 5 and 6: Pumping system Sample introduction
Page 7 and 8: Why is a vacuum necessary? The mean
Page 9 and 10: Various ionisation methods �� E
Page 11 and 12: Electrospray ionization 11/21/2005
Page 13 and 14: Droplet shrinks due to solvent evap
Page 15 and 16: Two examples of common ESI oxidatio
Page 17 and 18: The principal outcome of the electr
Page 19 and 20: ESI + + + + + Direction of electric
Page 21 and 22: ES spectrum of Rho protein 100 % 68
Page 23 and 24: 100 % ES spectrum after deconvoluti
Page 25 and 26: 22+ 11/21/2005 25 22+
Page 27 and 28: Matrix assisted laser desorption io
Page 29: A standard 100 well UV MALDI sample
Page 33 and 34: Why is the matrix so important? �
Page 35 and 36: Matrix Assisted Laser Desorption/Io
Page 37 and 38: Pumping system Sample introduction
Page 39 and 40: -(U + V o cosΩt) r r o + − −
Page 41 and 42: http://elchem.kaist.ac.kr/vt/chem-e
Page 43 and 44: What if we place three quadrupoles
Page 45 and 46: Product ion scan or MS/MS MS/MS gas
Page 47 and 48: MS Neutral loss scan gas Neutral lo
Page 49 and 50: Time-of Time of-flight flight (ToF)
Page 51 and 52: Reflectron ion optics: s3 s2 s1 700
Page 53 and 54: Time-Of Time Of-Flight Flight MS 11
Page 55 and 56: Ion traps: Hyperbolic 11/21/2005 55
Page 57 and 58: Ion Trap. Axial Radial toroidal rin
Page 59 and 60: Once in the trap 11/21/2005 59
Page 61 and 62: Ion traps: Linear 11/21/2005 61
Page 63 and 64: Ion traps: Orbital 11/21/2005 63
Page 65 and 66: Lord of the ion rings: Forging of t
Page 67 and 68: LTQ-Orbitrap: LTQ Orbitrap: 2 nd ge
Page 69 and 70: 700 L/s 250 L/s Ion path Q 760 Torr
Page 71 and 72: 11/21/2005 71
Page 73 and 74: GR q0 SL Pusher Q1 Puller q2 TOF Li
Page 75 and 76: Fourier transformed ion cyclotron 1
Page 77 and 78: Protein identification by mass spec
Page 79 and 80: Protein identification: Bioinformat
Page 81 and 82:
MALDI-TOF spectrum 11/21/2005 81
Page 83 and 84:
Proteomic applications of mass spec
Page 85 and 86:
Protein identification by mass spec
Page 87 and 88:
MEF2A MGRKKIQITRIMDERNRQVTFTKRKFGLM
Page 89 and 90:
Database Searching WWW Sites: Matri
Page 91 and 92:
AGAATVKENSSTRVKHCAAGAAWKAGGARERAAKT
Page 93 and 94:
Select: TNSDIVETIR m n R m 1 IR m 2
Page 95 and 96:
The peptide fragments give us seque
Page 97 and 98:
The 9 step strategy for manual inte
Page 99 and 100:
The 9 step strategy for manual inte
Page 101 and 102:
400 300 200 100 0 Step 2: b 2 and a
Page 103 and 104:
400 300 200 100 0 Step 4: Find y n-
Page 105 and 106:
400 300 200 100 0 Step 6: Find b-io
Page 107 and 108:
400 300 200 100 0 Peptide sequence:
Page 109 and 110:
Relative Abundance 100 80 60 40 20
Page 111 and 112:
Relative Intensity TOF Binary data
Page 113 and 114:
80 40 0 Identification of Proteins
Page 115 and 116:
A) B) C) ∆p Sample introduction:
Page 117 and 118:
A) B) C) HPLC/ autosampler Sample i
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Mass Spectrometry - Faculté de médecine de l'université d'Ottawa

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