MATRIX ASSISTED LASER

DESORPTION/IONIZATION TIME OF FLIGHT

(MALDI-TOF)

Submitted to: Submitted by:

Dr. Rajeev Kapoor Yashika Rani
Associated Professor PhD coursework (2293)
Department of Microbiology Department of Microbiology
 MALDI-TOF is a mass spectrometry technique used for the analysis of Proteins,
Peptides, and other biomolecules.

Basic mechanism

Sample is mixed with matrix solution which acts as laser absorbing agent and helps in ionizing the
molecules. The mixture is then spotted onto a target plate and dried and a laser is used to vaporise the
sample. The resulting ions are then accelerated through a time of flight tube, which separate the ions
based on their mass to charge ratio. The ions detected at the end of tube, and the resulting spectrum is
used for the identification and quantification of the sample molecules.
 STEPS

 Sample preparation :
Sample is prepared by mixing it with a matrix solution, which absorbs the laser energy and helps in ionizing
the molecules.
The sample and matrix are co-crystallized on a target plate, and the mixture is allowed to dry.
 Laser desorption/ionization :
A pulsed laser is used to vaporise the matrix and the sample, which leads to the desorption and ionization of
the molecules.
The laser energy is absorbed by the matrix, which generates a plume of ions from the sample.
 Ion acceleration :
The ions produced by the laser are accelerated through a vacuum tube using an electric field.
The ions are separated based on their mass to charge ratio as they travel through the tube, with lighter ions
travelling faster than heavier ones.
 Time of Flight analysis :
The separated ions reach a detector at the end of the tube, and the time it takes for the ions to travel from the
ion source to the detector is recorded.
This is Time of Flight and it is used to determine the mass of each ion.
The resulting mass spectrum is used for the identification and quantification of the sample molecules.
 Data analysis :
The mass spectrum obtained from the TOF analyser is analysed using specialized software, which compares
the peaks in the spectrum to a database of known mass spectra.
The software can identify the sample molecules and provide information about their molecular weight,
structure, and composition.
 TYPES

 Linear MALDI-TOF –
This is the most common type of MALDI-TOF mass spectrometer.
It uses a linear TOF analyser which separates the ions based on their mass to charge ratio and time of
flight in a straight line.
 Reflection MALDI-TOF –
It uses a reflectron to improve the resolution and accuracy of the mass analysis.
The reflectron is a curved electrostatic field that reflects the ions back towards the ion source, allowing
for longer flight time and improved resolution.
 MALDI-TOF/TOF –
This is a tandem mass spectrometry technique that combines MALDI-TOF with a second TOF analyser.
It allows the fragmentation of the ions in the first TOF analyser and mass analysis of the fragment ions in
the second TOF analyser.
This prides more detailed information about the composition of the sample molecules.
 MALDI-TOF/MS/MS –
It combines MALDI-TOF with two additional mass spectrometry analyser.
It allows for even greater fragmentation and analysis of the sample molecules, providing more
detailed structural and compositional information.

The type of MALDI-TOF mass spectrometer used depends

on the specific application and analytical requirements of the
sample being analysed.
APPLICATION

o Proteomics : to identify and quantify protein in complex mixture.

o Clinical microbiology : rapidly identify the microorganism such as bacteria and fungi.
o Drug discovery : to screen and identify potential drug target and lead compounds. It can
also be used to monitor metabolism and pharmacokinetics of drug.
o Environmental analysis : identify and analyse the pollutants and other environmental
contaminants. It can also be used to study the microbial communities in soil and water.
o Food safety : identify and quantify food borne pathogen.
THANK YOU