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MALDI-TOF
Mass
Spectrometry
Introduction
❖ MALDI is the abbreviation for "Matrix
Assisted Laser Desorption/Ionization.”
❖ MALDI is appropriate to analyze
biomolecules like peptides, lipids,
saccharides, or other organic
macromolecules.
❖ TOF stands for “Time of Flight”.
❖ This generates characteristic mass
spectral fingerprints which is compared
with large library of mass spectra.
❖ A versatile analytical technique to detect
and characterize mixtures of organic
molecules.
Principle of MALDI
❖ Matrix absorbs the ultraviolet light (nitrogen laser light, wavelength 337 nm) and converts it
to heat energy.
❖ The analyte is embedded on the Target Plate.
❖After a laser pulse, the irradiated spot is rapidly heated and becomes vibrationally excited.
❖Matrix molecules energetically ablated from the surface of the sample.
❖During ablation, the analyte molecules are usually ionized by being protonated or
deprotonated with the nearby matrix molecules.
Types of MALDI Variation Commonly
Used
❖ Laser: ultraviolet (UV) lasers are by far the most
important light sources.
❖ Time Of Flight: ideally suited to the MALDI ionization
process since the pulsed laser takes individual 'shots'
rather than working in continuous operation.
❖ Atmospheric Pressure: is an ionization technique (ion
source) that operates at normal atmospheric
environment.
TOF analyser
❖ The mass analyser used for MALDI is TOF analyser.
❖ It allows accurate mass determination for high molecular
weight species
❖ It makes TOF ideal for biomolecules.
❖ The ions desorbed by the laser pulse for accelerated in
an electric field to a kinetic energy of several KeV.
TOF Analyser
Accelaration
distance
Field Free Drift Distance
Electrode with gridSample
holder Detector
TOF Analyser
❖ Flight tube length and flight time
❖ At the end of the tube, ions hit the detector. And
is measured electronically.
❖ Lighter ions are accelerated more than the heavier ions
and reach the detector first.
2 m nanoseconds
TOF analyser
❖ Kinetic energy of the drifting ions is defined as
❖ 𝐸 𝑘𝑖𝑛 =
1
2
. 𝑚. 𝑣2
= 𝑧. 𝑒. 𝑉
❖ The velocity can be defined as
❖ 𝑣 =
𝐿
𝑡
❖ Substituting the velocity in eq.
❖
𝑚
𝑧
=
2.𝑒.𝑉
𝐿2 𝑡2
Sample pre-treatment
❖ Sample excess matrix.
co-crystallised
Inert
Stable in
vacuum
Embed with
analyte
Destruction
Fragmentation
Co-desorption of the analyte
sample treatment
• Dried droplet method
Mix Dry
Sample treatment
• Fast evaporation method
Apllications of Maldi
❖ Used for the analysis of proteins and peptides .
changes in structure can also be identified.
❖ Vey low amount of sample . sample preparation is
easy and spectra is simple.
❖ Very fast separation method
❖ https://youtu.be/0jeFpXHZ8W0

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MALDI-TOF Mass Spectrometry

  • 2. Introduction ❖ MALDI is the abbreviation for "Matrix Assisted Laser Desorption/Ionization.” ❖ MALDI is appropriate to analyze biomolecules like peptides, lipids, saccharides, or other organic macromolecules. ❖ TOF stands for “Time of Flight”. ❖ This generates characteristic mass spectral fingerprints which is compared with large library of mass spectra. ❖ A versatile analytical technique to detect and characterize mixtures of organic molecules.
  • 3. Principle of MALDI ❖ Matrix absorbs the ultraviolet light (nitrogen laser light, wavelength 337 nm) and converts it to heat energy. ❖ The analyte is embedded on the Target Plate. ❖After a laser pulse, the irradiated spot is rapidly heated and becomes vibrationally excited. ❖Matrix molecules energetically ablated from the surface of the sample. ❖During ablation, the analyte molecules are usually ionized by being protonated or deprotonated with the nearby matrix molecules.
  • 4.
  • 5. Types of MALDI Variation Commonly Used ❖ Laser: ultraviolet (UV) lasers are by far the most important light sources. ❖ Time Of Flight: ideally suited to the MALDI ionization process since the pulsed laser takes individual 'shots' rather than working in continuous operation. ❖ Atmospheric Pressure: is an ionization technique (ion source) that operates at normal atmospheric environment.
  • 6. TOF analyser ❖ The mass analyser used for MALDI is TOF analyser. ❖ It allows accurate mass determination for high molecular weight species ❖ It makes TOF ideal for biomolecules. ❖ The ions desorbed by the laser pulse for accelerated in an electric field to a kinetic energy of several KeV.
  • 7. TOF Analyser Accelaration distance Field Free Drift Distance Electrode with gridSample holder Detector
  • 8. TOF Analyser ❖ Flight tube length and flight time ❖ At the end of the tube, ions hit the detector. And is measured electronically. ❖ Lighter ions are accelerated more than the heavier ions and reach the detector first. 2 m nanoseconds
  • 9. TOF analyser ❖ Kinetic energy of the drifting ions is defined as ❖ 𝐸 𝑘𝑖𝑛 = 1 2 . 𝑚. 𝑣2 = 𝑧. 𝑒. 𝑉 ❖ The velocity can be defined as ❖ 𝑣 = 𝐿 𝑡 ❖ Substituting the velocity in eq. ❖ 𝑚 𝑧 = 2.𝑒.𝑉 𝐿2 𝑡2
  • 10. Sample pre-treatment ❖ Sample excess matrix. co-crystallised Inert Stable in vacuum Embed with analyte Destruction Fragmentation Co-desorption of the analyte
  • 11. sample treatment • Dried droplet method Mix Dry
  • 12. Sample treatment • Fast evaporation method
  • 13. Apllications of Maldi ❖ Used for the analysis of proteins and peptides . changes in structure can also be identified. ❖ Vey low amount of sample . sample preparation is easy and spectra is simple. ❖ Very fast separation method