Hard vs.

Soft Ionization methods,

ICP sources
Spark Sources
Glow Discharge
Secondary Ion/Neutral Mass Spec
fast atom bombardment (FAB) sources
desorption sources
field ionization (FI)
field desorption,
laser desorption . . . (especially MALDI)
plasma desoprtion sources
electron impact (EI)
chemical ionization (CI)
electrospray ionization sources (ESI)
Desorption electrospray ionization (DESI)
ICP sources
gas, liquid or solid
sample is introduced
into hot plasma
an efficient source
of positively charged
analyte ions
Ar plasma is
generated and maintained at the end of the glass torch located
inside the loops of a water cooled copper load coil.
RF potential applied to the coil produces an electromagnetic
field in the part of the torch located within its loops.
electrons are accelerated and collide with Ar atoms in the Ar
gas flowing through the torch producing Ar
+
ions and free
electrons - a plasma.
ICP sources
the ions have to be extracted
from the high temperature
(~ 6000K or more), atmospheric
pressure (760 torr) environment
of an often chemically corrosive
Ar plasma into a mass spectrometer
operating in a high vacuum (10-5 torr) at room temperature.
interface region contains two successive cones (mm orifices)
ions in the center of the plasma are sampled into the region
between two cones held at a pressure of about 1-3 torr
At this stage, most of the Ar atoms are removed by a vacuum pump.
ion beam is further extracted through the skimmer cone orifice
into the front section of the mass spectrometer (pressure of
about 10
-3
- 10
-4
torr)
Spark Ionization Sources
samples are physically incorporated into two conductive
electrodes (usually either carbon or silver)
a high-voltage arc is produced, ionizing the material
semiquantitative trace element technique for solids and liquids
samples: conducting, semiconducting and insulating solids,
powders, crystals, liquids, organometallics, ash from organics,
unknowns and many other sample forms.
detection capabilities encompass the periodic table (Li U)
has the ability to determine impurity levels from the sub-ppm
level to 0.1%. SSMS
total simultaneous elemental coverage
low detection limits
high res. capabilities - eliminates many spectral interferences.
Glow Discharge MS
analytical technique for the
bulk elemental analysis of
inorganic solid samples.
capable of analyzing,
conducting, semi-conducting
and insulating samples.
amenable to solids, powders,
crystals, wafers, and many other sample forms.
elemental coverage encompasses Li U
determine impurity levels from the sub-ppb to percent level
advantages include
high precision and low detection limits,
quantitative accuracy (+/- 25% on average), without the use of standards
high resolution capabilities eliminate most spectral interferences.
Secondary Ion/Neutral Mass Spec
a primary, high-energy beam
of ions (usually oxygen,
argon, or cesium) is aimed
at a small area of a sample,
such as a mineral grain.
the primary ions have
energies of ~ 10 keV
the primary ions sputter
away the sample by causing
the ejection of atoms and
ions (called secondary
neutrals and ions)
these secondary ions
(approximately 1% of the
sputtered material) are
accelerated into a mass
spectrometer to reveal the
elemental and isotopic
characteristics of the sample.
Fast Atom Bombardment (FAB)
material to be analyzed is
mixed with a non-volatile
chemical protection
environment called
a matrix
This is bombarded under
vacuum with a high energy
(4 10 keV) beam of atoms.
atoms are typically an inert gas (Ar or Xe)
common matricies include glycerol, thioglycerol,
3-nitrobenzyl alcohol (3-NBA), 18-Crown-6 ether,
2-nitrophenyloctyl ether, sulfolane, diethanolamine,
and triethanolamine.
Field Desorption
Field desorption (FD) is a method for emitting ions into the
gas phase.
Sample spread on an emitter is heated while a high electric
field is applied.
Ions are then emitted by the tunneling, ion-molecule reactions,
thermal fusion effects,
and other phenomenon
occurring on the emitter
surface and around the
whisker ends.
The ionization phase depends strongly on the sample material
and the spread condition.
Plasma Desorption
Plasma desorption ionization mass spectrometry (PDMS; also
called fission fragment ionization) is a mass spectrometry
technique in which ionization of material in a solid sample by
bombarding it with ionic or neutral atoms formed as a result of
the nuclear fission of a suitable nuclide, typically the
Californium isotope
252
Cf
Californium-252 plasma desorption mass spectroscopy
RD Macfarlane and DF Torgerson
We have shown that 252Cf-PDMS is capable of producing mass spectra of
quasi-molecular ions for a wide variety of compounds, including amino
acids, moderately large peptides, nucleotides, and natural products. Positive
and negative ion mass spectra can be obtained, and in many cases quasi-
molecular ions are observed in both. The method is nondestructive, as only
a relatively few molecules are used and samples can be recovered after the
measurement is made. Fragmentation patterns are obtained which can yield
structure information. The present sensitivity of the method is at the
nanogram level and there are possibilities for reducing this to picograms.
The mass resolution is sufficient to give elemental identification up to mass
500. This may be extended to higher masses with improved time-of-flight
techniques. There are indications that 252Cf-PDMS may extend the mass
range of molecules that can be studied to as high as 3000 or more.
Science, Vol 191, Issue 4230, 920-925
Copyright 1976 by American Association for the Advancement of Science
Especially MALDI
Matrix-assisted laser desorption ionization (MALDI)
Ionization method using matrix-assisted laser desorption.
a soft ionization technique
analysis of biomolecules
proteins,
Peptides
sugars)
large organic molecules
polymers,
dendrimers
other macromolecules
tend to be fragile and
fragment when ionized
by other methods.
MALDI contd
identity of suitable matrix compounds is determined using
specific molecular design considerations
fairly low molecular weight (to allow facile vaporization)
large enough (with a high enough vapor pressure) not to evaporate
during sample preparation or while standing in the spectrometer
are acidic / act as a proton source to encourage ionization of the analyte
have strong absorption in the UV so they rapidly and efficiently absorb
the laser irradiation
functionalized with polar groups - allowing use in aqueous solutions
matrix solution is mixed with the analyte (e.g. protein-sample)
organic solvent allows hydrophobic molecules to dissolve
water allows for hydrophilic molecules to do the same
solution is spotted onto a MALDI plate
solvents vaporize, leaving only the recrystallized matrix
analyte molecules spread throughout the crystals in co-crystallized
MALDI spot
laser desorption . . .
UV MALDI Matrix List
Compound Solvent l (nm) Applications
2,5-dihydroxy benzoic
acid
acetonitrile, water,
methanol, acetone,
chloroform
337, 355,
266
peptides,
nucleotides,
oligonucleotides,
oligosaccharides
3,5-dimethoxy-4-
hydroxycinnamic acid
acetonitrile, water,
acetone, chloroform
337, 355,
266
peptides, proteins,
lipids
4-hydroxy-3-
methoxycinnamic
acid
acetonitrile, water,
propanol
337, 355,
266
proteins
-cyano-4-
hydroxycinnamic acid
acetonitrile, water,
ethanol, acetone
337, 355
peptides, lipids,
nucleotides
Picolinic acid Ethanol 266 oligonucleotides
3-hydroxy picolinic
acid
Ethanol 337, 355 oligonucleotides
Field Ionization
Field ionization (FI) is the generation of M+ ions by removal
of electrons, primarily from gas sample molecules, using a
high electric field.
This generally occurs at a sharp edge or tip that is biased to a
high electrical potential
Field ionization (FI) is a method that uses very strong electric fields to produce ions
from gas-phase molecules. Its use as a soft ionization method in organic mass
spectrometry is principally due to Beckey [8]. Like EI or CI, FI is only suitable for gas-
phase ionization.
Therefore, the sample is introduced into the FI source by the same techniques that
are commonly used in EI and CI sources, for example using a direct probe that can be
heated or the eluent from a gas chromatograph.
The intense electric fields used in this ionization method are generally produced by a
potential difference of 812 kV that is applied between a filament called the emitter
and a counter-electrode that is a few millimetres distant. Sample molecules in the gas
phase approach the surface of the emitter that is held at high positive potential. If the
electric field at the surface is sufficiently intense, that is if its strength reaches about
107108 Vcm1, one of the electrons from the sample molecule is transferred to the
emitter by quantum tunnelling, resulting in the formation of a radical cation M+.
This ion is repelled by the emitter and flies towards the negative counter-electrode. A
hole in the counter-electrode allows the ion to pass into the mass analyser
compartment. In order to achieve the high electric field necessary for ionization, the
emitter constituted of tungsten or rhenium filament is covered with thousands of
carbon microneedles on its surface. It is at the tips of these microneedles that the
electric field strength reaches its maximum. FI leads to the formation of M+ and/or
MH+ ions depending on the analyte.
The formation of protonated molecular species results from ionmolecule reactions
that can occur between the initial ion and the sample molecules close to the surface of
the emitter. It is not unusual to observe both M+ and MH+ in the FI spectrum.
Electron Ionization (EI)
EI (formerly known as electron impact) is an ionization
technique widely used in mass spectrometry, particularly
for organic molecules.
The gas phase reaction producing electron ionization is:
M + e
-
M
+
+ 2e
-

low energies (around 20 eV), the interactions between the
electrons and the analyte molecules do not transfer enough
energy to cause ionization
at around 70 eV, the de Broglie wavelength of the electrons
matches the length of typical bonds in organic molecules (about
0.14 nm), and energy transfer to organic analyte molecules is
maximized, leading to the strongest possible ionization and
fragmentation
Electron Ionization (EI)
Chemical Ionization (CI)
Chemical ionization (CI) is an ionization technique used in
mass spectrometry
ionization is achieved by interaction of its molecules with
reagent ions
the analyte is ionized by chemical ion-molecule reactions
during collisions in the source
the process may involve transfer of an electron, a proton or
other charged species between the reactants.
a less energetic procedure than electron ionization and the ions
produced are, for example, protonated molecules: [M + H]+.
These ions are often relatively stable, tending not to fragment
as readily as ions produced by electron ionization.
Chemical Ionization (CI)
typical reagent gases (ex. CH
4
, isobutane, or NH
3
) are present
in a millionfold excess with respect to the analyte.
analyte is ionized by ion-molecule chemical reactions:
Primary Ion Formation:
CH
4
+ e
-
CH
4
+
+ 2e
-
Secondary Reagent Ions:
CH
4
+ CH
4
+
CH
5
+
+ CH
3
CH
4
+ CH
3
+
C
2
H
5
+
+ H
2
Product Ion Formation:
M + CH
5
+
CH
4
+ [M + H]
+
(protonation)
AH + CH
3
+
CH
4
+ A
+
(H

abstraction)
M + CH
5
+
[M+ CH
5
]
+
(adduct formation)
A + CH
4
+
CH
4
+ A
+
(charge exchange)
Electrospray Ionization Sources (ESI)
Electrospray ionization (ESI) was first introduced by Dole and
coworkers in 1968 and coupled to MS in 1984 by Yamashita
and Fenn
In ESI the sample is dissolved in a polar, volatile solvent, and
transported through a needle placed at high positive or
negative potential (relative to a nozzle surface)
The high electric potential (1 to 4 kV) between the needle and
nozzle causes the fluid to form a Taylor cone, which is
enriched with positive or negative ions at the tip. A spray of
charged droplets is ejected from the Taylor cone by the electric
field. The droplets shrink through evaporation, assisted by a
warm flow of nitrogen gas passing across the front of the
ionization source (Fig. 2.6).

Mass Spektrometry, Dominic M. Desiderio & Nico M. Nibbering., Series Eddition.
Mass Spektrometry, Dominic M. Desiderio & Nico M. Nibbering., Series
Eddition.
Ions are formed at atmospheric pressure and pass
through a coneshaped orifice, into an intermediate
vacuum region, and from there through a small
aperture into the high vacuum of the mass
analyzer.

Sample preparation requires only dissolution of the sample to a
suitable concentration in a mixture of water and organic solvent,
commonly methanol, isopropanol, or acetonitrile. A trace of formic
acid or acetic acid is often added to aid protonation of the analyte
molecules in the positive ionization mode. In negative ionization
mode ammonia solution or a volatile amine is added to aid
deprotonation of the analyte molecules.
The sensitivity of ESI-MS is good, with low femtomole or attomole
detection levels for many peptides. However, the sensitivity of ESI
is a function of the concentration of the injected sample. High flow
rates, that is, 1 to 1000 mL/min in conventional ESI-MS, result in
high sample consumption. It is therefore advantageous to use the
lowest possible flow rate. Nano-ESI (or nanospray) is a low-flow-
rate (20 to 200 nL/min) version of ESI, with lower sample
consumption and considerably higher sensitivity.
ESI is an soft ionization method, accompanied by very little
fragmentation of the formed molecular ions. Consequently,
weak bonds are often preserved and analysis of intact post-
translationally modified peptides/proteins and noncovalently
bound complexes, such as proteinligand complexes, can be
successfully performed with ESI-MS.
Desorption Electrospray Ionization
Desorption ESI (DESI) was introduced by Takatz et al. The
method is sensitive and large species such as proteins can be
detected. The ions observed are more or less the same as with
regular ESI.
A DESI source consists of a spray capillary and a coaxial
capillary providing the nebulizer gas. High voltage is applied
to the spray needle, which is directed towards the target
surface (Fig. 2.7).
Mass Spektrometry, Dominic M. Desiderio & Nico M.
Nibbering., Series Eddition.
Sample species are then desorbed and will subsequently enter
the orifice to the mass spectrometer. Normal distances between
the spray needle, sample, and orifice range from some
millimeters to several centimeters.
The optimum geometry depends on the sample and on the size
of the desired sampling area. The advantages with DESI are
that the target can be in principle any type of surface and that
the analysis time often can be very short, on the order of
seconds. This means that rapid analyses can be performed
without the need for sample preparation.