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

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Advantages �� MALDI primarily creates singly charged ions [M+H] + �� Less sensitive to contaminants �� Sensitivity at femtomole level �� High throughput analysis Brancia FL, Trieste, 12/02/2004 11/21/2005 36
Pumping system Sample introduction DIRECT INTRODUCTION (solid, liquid, gas) SEPARATION TECHNIQUES (HPLC, CE, GC) ION SOURCE (“ion ion generation”) generation EI, FAB, MALDI,Electrospray TOF, quadrupole, quadrupole, ion trap vacuum ANALYZER (“mass mass analysis”) analysis Detector Data Processing Brancia FL , Trieste, 12/02/2004 11/21/2005 37
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 and 30: A standard 100 well UV MALDI sample
Page 31 and 32: Sample plate 2-3 kV + + + + + + + +
Page 33 and 34: Why is the matrix so important? �
Page 35: Matrix Assisted Laser Desorption/Io
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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