Mass spectrometry

For what purposes MS is used?

▶ For measuring the molecular mass of a sample.
▶ For identification of the unknown compounds little as 10-
12g, 10-15 moles ; compounds identified at very low

concentrations (one part in 1012) in chemically complex

mixtures.
▶ For quantification of known compounds.
▶ Used to discover the number of isotopes ,determine the
relative abundance of the isotopes and measure of their
exact masses.
▶ Within an accuracy of 0.01% of total weight of sample
and within 5 ppm for small organic molecules
▶ helps to determine the chemical and structural
information about molecules.
▶ Fragment sample & analyse products.
▶ Uses a machine called Mass Spectrometer.
 Electron Ionization (EI) technique

EI is the most appropriate technique for relatively small (m.w.<600) neutral organic
molecules which can easily be promoted to the gas phase without decomposition, i.e. volatile.
Since EI samples are thermally desorbed to the gas phase and subjected to the high energy of
EI, analytes must be thermally stable. The gas phase molecules enter into the ion source
where they are bombarded with free electrons emitted from a filament (Figure 1). The
electrons bombard the molecules causing a hard ionization that fragments the molecule, and
turn into positively charged particles called ions.
MALDI | Matrix Assisted LASER Desorption Ionization Technique
It is a soft ionization technique under desorption ionization methods, which
uses pulsed LASER (Light Amplification by stimulated Emission of Radiation)
beam.

It is used to determine the molecular weight of peptides, antibodies,

protiein molecules etc. up to the size of 300 Kda.

Matrix compound α-cyano-4-hydroxycinnamic acid (CHCA),

2,5-dihydroxybenzoicacid (DHB), 2-(4-hydroxyphenylazo) benzoic acid
(THAP), sinapinicacid (SA) and 2-(4-hydroxy-phenylazo) benzoicacid (HABA).