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MALDI TOF MS

Muhammad Farooq

1/31/2013

Institut fr Physik

OUTLINE
What Is MALDI-TOF MS?
Fundamentals of MALDI-TOF MS

Sample Preparation
MALDI TOF MS In Action

Example

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Institut fr Physik

Mass Spectroscopy
An analytical spectroscopic tool primarily concerned with the separation of
molecular (and atomic) species according to their mass

Configuration of the mass spectrometer


Inlet

Ion source

Mass Analyzer

Detector

Sample plate Target Solids probe

MALDI Electro-spray APCI EI CI

TOF Quadrupole Ion Trap Magnetic Sector FTMS

Micro-channel PCs Plate DEC Station Electron Multiplier Hybrid


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Problem With Polymers


Polymer Characterization With Mass Spectroscopy Is Not Possible Reason !!
MS Requires Gas Phase Ions Polymers Composed Of Large, Entangled Chains Not Easily Converted To Gas Phase Ions
Thermal Energy Information Loss

Decomposition

Fragmentation

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Institut fr Physik

MALDI-TOF MS
Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry
The general principle of MALDI-TOF MS revolves around the rapid photovolatilization of a sample embedded in a UV-absorbing matrix followed by

time-of-flight mass spectrum analysis


Mass Range
Detection Limit Accuracy

1-400,000 Da
10-15 10-18 moles 0.1 0.01%

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Institut fr Physik

Fundamentals of MALDI -TOF MS


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Analyzer
Sample 1 Polymer along with UV absorbing matrix prepared by a special technique Laser Beam 2 Nitrogen Laser Pulse (337 Nm)/ UV Ion Source 3 Generating Of Gas-phase Ions From Solid Phase(sample) Flight Tube 4 Connecting Tube Between The Ion Source And Detector Detector 5 Determination of mass of ions, resolved by the mass analyzer
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Sample Preparation
MALDI Sample Matrix Polymer

Must be able to embed analyte molecules


Isolate the oligomers Absorb energy Desorb the analyte
Cyano Hydroxycinnamic Acid (CHCA) Sinapic Acid (SA) Dihydrobenzoic Acid (DHA) Hydroxypicolinic Acid (DHA)

Ionize the analyte


Ratio M:A is 1000:1 to 10000:1
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Institut fr Physik

Sample Preparation
Methods
Dried Droplet Method Layer Method Electrospray Matrix-pre Coated Layers Fast-evaporation Method Chemical Liquid Particle-doped (Two-phase) Liquid

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Institut fr Physik

Sample Preparation
Dried-Droplet
introduced in 1988 by Hillenkamp and Karas A drop of aqueous matrix compound solution is mixed with analyte solution A solid deposit of analyte-doped matrix crystal is obtained Sample placed into the mass spectrometer for analysis.

Matrix
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Polymer

Mix
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Sample
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MALDI TOF MS In Action


Pulsed Laser 337 Nm & 355 Nm Matrix / Analyte Polymer Matrix Detector

Flight Tube

MALDI Ion Focusing Lens

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Institut fr Physik

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Time-of-Flight Mass Analyzer


Linear Mode
Flight Tube

Ion Source

Field free drift tube

Detector

The lighter ions travel faster and strike the detector before the heavier ions. This time of flight (TOF) can be correlated with mass of the ion

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Institut fr Physik

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Time-of-Flight Equations
In the ideal case all ions leave the source with the same kinetic energy
U =kinetic energy of the ion V= applied potential

e = electron z = charge

The same kinetic energy accelerating the ions based on the mass-to-charge (m/z) ratios

Assuming that most of the ion's flight time (t) is spent in the drift tube (D>>s), the

time-of-flight can be determined through

conversion of the time-of-flight data into a direct mass-to-charge relation

=
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Mass Calibration

M3

M2

M1

M2

M1 M3

t1

t2

t3

from empirical determination of the times-of-flight of several known Masses, a calibration line is drawn, according to = +
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a & b are calibration constants

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Resolution
Resolution is the ability to distinguish two ions of different masses, and is usually scaled to the mass range under consideration, such that is the mass difference between two adjacent peaks that are just resolved Average mass Low resolution High resolution

/z
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Resolving Power Example


RP= 3,000
100 100

RP= 5,000
100

RP= 7,000 C6H5Cl C6H5OF

80

80

80

In ten sity (%)

In ten sity (%)

60

60

In ten sity (%)

60

40

40

40

20

20

20

0 111.95 112.00 Mass [amu] 112.05 112.10

0 111.95 112.00 Mass [amu] 112.05 112.10

0 111.95 112.00 Mass [amu] 112.05 112.10

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Example

MALDI-TOF MS spectra of epoxide-amine

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Institut fr Physik

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Thank You

1/31/2013

Institut fr Physik

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