You are on page 1of 13

MALDI-TOF Spectrometry

Stephen Gangemi November 2002

Overview

What is MALDI-TOF? How does it work? Why is it useful? What are its figures of merit? What is its future?

What is MALDI-TOF?

Matrix-assisted laser desorption/ionization time of flight mass spectrometry


Ionizes molecules via laser pulses Separates molecules according to mass to charge ratio

Mainly used for detection of large biomolecules

Development of MALDI-TOF
Developed in 1988 by Professor Hillenkamp Designed to enhance mass-spec. by solving two main problems

Thermal instability and low volatility Large and heavy biomolecules

Prof. Franz Hillenkamp

The Principles of MALDI

Dissolve compound and add matrix Evacuate air from sample chamber Laser shoots pulses of light at sample

TOF Spectrometer
Ions are accelerated via charge Separated by time it takes to reach detector

TOF Analysis the Math Behind It All

Equate accelerating potential to Kinetic energy

zV=1/2mv2

Substitute distance / time for v

zV= 1/2m(x/t)2 m/z=(2Vx2)/t2 m/z = A(B-t)2, where A and B are calibration constants
m=mass z=charge t=time V=accelerating potential x=length of chamber

For more precision use calibration standards

Factors Affecting Quality of Output

Size of crystals Energy of laser Detector voltage Accelerator voltage Extraction time Number of laser shots

Evaporation rate Sample concentration Baseline calculations Scanning settings

Uses of MALDI-TOF
Used to characterize and identify large molecules Used in pharmaceutical for QC, monitoring of enzyme reactions Used in DNA sequencing for forensics Used to identify different strains of viruses to help develop vaccines

Figures of Merit
Mass Range
Detection Limit Accuracy Sensitivity Sample Consumption

1-400,000 Da
10-15 10-18 moles 0.1 0.01% 1 uL containing 100 fmol 10 pmol Less than 1 pmol

Advantages / Disadvantages
Small sample needed Works well with large biomolecules Gives absolute mass measurements

Finding matrices to work with samples Ions may collisionally relax Sensitive to contaminants, such as salts

Whats its future?


Will help revolutionize the medical world and will help lead to treatments for many diseases Will be useful for DNA sequencing, thus can be useful for forensic investigations

Sources
http://www.psrc.usm.edu/mauritz/maldi.html

http://www.psrc.usm.edu/macrog/maldi.htm
http://www.metabion.com/techinfo/z-maltof.html http://cbms.st-andrews.ac.uk/services/maldi-tof/maldi.html http://www.lsc.psu.edu/stf/imsc/MaldiTof.html http://www.babraham.com/new/maldi.htm http://www.humboldt-forum-recht.de/6-1997/Biographie.html http://www.biotech.iastate.edu/facilities/protein/maldi.html

http://www.geocities.com/justinhettick/maldi.html
http://www.itl.nist.gov/div898/pubs/ar/ar1998/node13.html http://cmgm.stanford.edu/pan/voyager.html http://www.laserscience.com/maldi_tof.htm