MASS SPECTROMETRY/Principles and Instrumentation
Poulsen M (1995) Føroyskar Føðslutalvur (Faroese Food
Composition Tables.) Heilsufrøðiliga Starvstovan, pp.
1–25. Tórshavn: Food and Environmental Agency.
Skaale Ó and Johannesen M (1974) Matur og Matgerð.
Grønalı́ð. Tórshavn. (Faroese cookbook.)
Market Research See Food Acceptability: Affective Methods; Market Research Methods
MASS SPECTROMETRY
Contents
Principles and Instrumentation
Applications
Principles and Instrumentation
F A Mellon, AFRC Institute of Food Research,
Norwich, UK
Copyright 2003, Elsevier Science Ltd. All Rights Reserved.
Principles
0001 Mass spectrometry is an extremely sensitive and spe-
cific analytical technique, capable of providing quali-
tative and quantitative analytical data on nanomolar
to attomolar amounts of analyte. It has been applied
successfully to a very wide range of analytical prob-
lems in the food and nutrition sciences. Mass spec-
trometers have become far more accessible, cheaper
to purchase and operate, and easier to use, especially
over the last decade. This is mainly a consequence of
the advent of compact, bench-top instruments, usu-
ally coupled to gas or liquid chromatographs. The
following article describes the basic theory of mass
spectrometry and the principal types of mass spectro-
metric equipment relevant to food and nutrition
analysis and research. Applications of mass spectrom-
etry and techniques of combined chromatography
mass spectrometry are covered in separate articles.
Mass Spectrometer
0002 All mass spectrometers comprise four main compon-
ents. First, an ion source, where sample molecules
may be ionized by a variety of means, second, a
mass analyzer that separates ions according to
their mass-to-charge ratio, m/z, third, a detector that
measures the abundances of the separated ions as an
electrical signal, and fourth, a recording device that
3739
Small-Type Coastal Whaling in Japan (1988) Report of
an International Workshop. Occasional publication
number 27. Alberta, Canada: Japan Social Sciences
Association of Canada Fund to Promote International
Educational Exchange and Boreal Institute for
Northern Studies.
converts the detector signal into a form suitable for
further study and processing.
Ionization Techniques – Organic Mass
Spectrometry
Ionization of organic molecules may be accomplished 0003
by a variety of techniques. The most common prac-
tical methods are described below.
Formation of Ions
Organic mass spectrometry 0004
Electron ionization (EI) The analyte is vaporized
into the EI source and bombarded with 70-eV elec-
trons. The ion source chamber is maintained at a
vacuum of 103 Pa. A proportion of the energy of the
electron beam is transferred to the molecule, ejecting an
electron to generate a molecular ion (Mþ ), the most
important ion in the mass spectrum (eqn (1)).
M þ e ! Mþ þ 2e: ð1Þ
Excess electronic energy is also transferred to the 0005
molecule during the ionization process and is rapidly
converted to internal vibrational energy. This energy
may be sufficient to induce bond cleavage in some
ions, generating fragment ions that may also decom-
pose. This is shown in the sequence of decompos-
itions of a hypothetical molecule ABCD (eqn (2)).
ABCDþ ! ABCþ þ D ! ABþ þ C ! Aþ þ B:
ð2Þ
Rearrangement processes may also occur (eqn (3)). 0006
ABCDþ ! ADþ þ CB: ð3Þ
 3740 MASS SPECTROMETRY/Principles and Instrumentation

0007 Most ions formed by EI have a single positive
charge (z ¼ 1), and m/z is then equivalent to the
mass of the ion. The way in which a molecule frag-
ment is determined by a number of factors that will
not be discussed here. Semiempirical rules for predict-
ing or rationalizing bond cleavages and rearrange-
ments can be found in several textbooks. The most
important consequence of the fragmentation process
is that it generates a characteristic, reproducible mass
spectrum that serves as a spectrometric ‘fingerprint’
of the analyte.
0008 Mass spectra are generally recorded in the form of
a line diagram (bar graph). Ion intensities are normal-
ized so that the largest peak in the mass spectrum, the
base peak, has an intensity of 100 units, and all other
ion abundances are expressed as a percentage of this
value. The low-resolution mass spectrum of caffeine
is shown in Figure 1.
0009 A molecular ion can be seen at m/z 194, and a
number of abundant fragment ions are also present.
In this example, the molecular ion is also the base
peak, although this is not always the case; strongly
favored fragmentations may yield abundant fragment
ions that are more intense than the molecular ion.
0010 Some molecules are difficult to analyze by EI mass
spectrometry for either or both of two main reasons.
They may fragment very readily, so that the molecular
ion is of very low abundance and therefore (in the
case of an unknown) difficult to assign. Alternatively,
the molecules may be involatile or thermally labile
and difficult or impossible to vaporize without
inducing thermolytic decomposition. Several ‘soft’
ionization methods are available for analyzing mol-
ecules that fall into this category. These techniques
are used to transfer analytes from the liquid or solid
phase to the gas phase and ionize them by mechan-
isms that impart minimal amounts of energy.
Chemical ionization (CI) CI is based on the reac- 0011
tions of gaseous analyte molecules with an excess of
ions formed by electron bombardment of a reagent
gas. These reactions take place at high (relative to EI)
ion source pressures, typically in the range 10–150 Pa.
Typical reagent gases include methane and ammonia:
these generate CHþ þ
5 and NH4 ions, respectively under
CI conditions. The reagent ions are present in large
excess (104:1) relative to analyte molecules. The re-
agent ions generally react with analyte molecules M
by protonation (eqn (4)),
M þ NHþ þ
4 ! MH þ NH3 : ð4Þ
Negative ion chemical ionization is also possi-
ble. Negative ion reagent gases include water. This
generates OH ions and reacts by proton abstraction
(eqn (5)),
M þ OH ! ½M  H  þ H2 O: ð5Þ
A special case of negative ionization, electron capture
negative chemical ionization (ECNCI) is not strictly
CI. It is an electron capture process where low-energy
electrons generate radical anions (eqn (6)),
M þ eðthermalÞ ! M : ð6Þ

194
100

O CH3
H3C N
Relative intensity (%)

N
N
109 O N
50
CH3
55

82

165

40 80 120 160 200

m/z ratio

fig0001 Figure 1 Low-resolution electron ionization mass spectrum of caffeine in bar graph format.
 MASS SPECTROMETRY/Principles and Instrumentation
In this case, the ‘reagent’ gas acts as a moderator,
slowing the electrons to the thermal energies where
the electron capture process become efficient. ECNCI
is particularly useful for determining analytes with
strong electron capturing properties, for example,
halogenated molecules such as dioxins. ECNCI can
yield much greater sensitivities and lower detection
limits in these cases.
0012 Positive and negative reagent ions may also react in
other ways. A range of gases are available for specific
applications. CI spectra usually contain intense pro-
tonated or negatively charged molecules and fewer
fragment ions than EI.
0013 Desorption EI and CI In conventional EI and CI,
analyte molecules are typically introduced into the
ion source via a heated probe or GC inlet. Desorp-
tion EI and desorption CI provide techniques for
obtaining mass spectra of less volatile, thermally
labile molecules by rapid heating whilst the samples
are very close to the electron beam or are immersed in
the CI plasma. These techniques are still used occa-
sionally but have largely been superseded by more
robust and efficient methods for analyzing involatile
molecules, as described below.
0014 Fast atom bombardment (FAB) and liquid secondary
ionization mass spectrometry FAB and liquid
secondary ionization mass spectrometry essentially
describe the same technique, and the terms can be
used interchangeably. FAB was the first ionization
technique to enjoy widespread success in analyzing
polar, labile, and large molecules, including peptides
and polysaccharides. The analyte in a viscous, low-
boiling liquid matrix, typically glycerol that is bom-
barded with a stream of atoms or ions, typically Xe
atoms or Csþ ions that have average translational
energies in the keV range. Abundant protonated
or cationized molecules, for example [M þ H]þ or
[M þ Na]þ and deprotonated ions, [M  H], are
formed and detected in positive and negative
ion modes, respectively. Characteristic fragment ions
may also be formed.
0015 FAB enjoyed great and deserved popularity in the
1980s and helped to establish mass spectrometry very
strongly in the biological sciences. However, it has
generally been superseded by the more powerful, ver-
satile techniques of electrospray and matrix-assisted
laser desorption ionization.
0016 Electrospray ionization (ESI) and atmospheric pres-
sure chemical ionization (APCI) In ESI, the analyte
is dissolved in a suitable solvent, for example a 50:50
mixture of acetonitrile and water, and is pumped
through a narrow diameter stainless steel capillary
3741
tube that is maintained at a voltage in the range of 1–
4 kV. This process generates a plume of charged liquid
droplets in atmosphere (the Taylor cone). A heated
(usually nitrogen) gas aids evaporation of the charged
droplets and helps break up clusters of analyte and
solvent ions. Analyte ions are generated from the
charged droplets by an evaporation mechanism. The
ions are conducted into the mass spectrometer vacuum
via a pumped nozzle-skimmer system (Figure 2).
A curtain of nitrogen gas, which also aids evapor- 0017
ation of the charged droplets, prevents cluster ion
formation. The ions are sampled through an orifice
and enter the mass analyzer. ESI has revolutionized
mass spectrometry by enabling mass spectra to be
obtained routinely on polar, involatile molecules.
Even high-molecular-weight biomolecules can be
analyzed by ESI. Proteins, for example, yield a series
of multiply charged ions well within the mass range of
conventional mass spectrometers such as quadrupoles
or magnetic sector instruments. These signals can be
transformed mathematically to yield the molecular
weight of the sample (Figure 3).
APCI is closely related to ESI and often employs an 0018
almost identical ion source. The only changes are the
addition of a discharge electrode and replacement of
the ESI spray probe with an APCI probe. In APCI,
flowing liquid, typically an HPLC eluent, that con-
tains the dissolved analyte(s) is conducted to a pneu-
matic nebulizer, where it forms a spray. As with ESI,
the spray is generated in atmosphere and is directed
towards a discharge electrode that is maintained at
1–4 kV. This is placed close to a small-diameter orifice
that leads to the high-vacuum mass analyzer region. A
pumped nozzle and skimmer arrangement removes
excess solvent molecules, and desolvated ions are
admitted to the mass analyzer, in a similar manner
to an ESI source. Reagent ions are generated from
solvent molecules in the electrical discharge near the
corona pin. These react with analyte molecules to
generate protonated or deprotonated molecules
([M þ H]þ or [MH]), in a similar manner to con-
ventional CI.
APCI is more robust (being less prone to solvent or 0019
solvent modifier effects) than ESI but is usually too
vigorous and energetic to yield useful data on fragile
biomolecules. However, it is very useful for determin-
ing molecules of low to intermediate polarity and is
capable of producing useful data on some quite polar
molecules, for example monoglycosides.
Matrix-assisted laser desorption Ionization 0020
(MALDI) MALDI is one of the best techniques for
obtaining analytical data, including molecular
weight information, on medium and large biomole-
cules. The sample, in low concentration, is mixed
 3742 MASS SPECTROMETRY/Principles and Instrumentation
N2 drying gas
Atmospheric pressure region
LC or Syringe pump flow
3.5 kV
Spray
Nozzle
fig0002 Figure 2 Schematic diagram of an electrospray ionization source.
with a concentrated solution of, for example, sinapi-
nic acid, on a metal target. The solution is dried, and
the target is introduced into the mass spectrometer
ion source vacuum. The sample plus matrix is irradi-
ated with a pulsed laser (typically a 327-nm nitrogen
laser). The matrix chosen should be sublimable and
be able to absorb and transfer energy at the laser
wavelength. The mechanism of desorption and ion-
ization is complex. It is thought that the matrix then
transfers sufficient transverse vibrational energy to
the sample molecules to desorb some of them with
very little excess energy for fragmentation. The tech-
nique is particularly useful for mass measuring large
and medium-sized biomolecules, especially peptides,
proteins, and DNA fragments, and generates
[MþH]þ, [MH] or adduct ions, [MþNa]þ.
0021 Other ionization methods Several other methods
have been used to ionize organic molecules in the
past. For example, field ionization, field desorption,
and 252Cf plasma desorption were once popular tech-
niques for analyzing otherwise intractable molecules,
such as biopolymers. These techniques are now
mainly of historical interest and have generally been
superseded by more practical, reliable and robust
methods, particularly ESI and MALDI.
0022 Inductively coupled plasma ion source (ICP-
MS) Inorganic mass spectrometry has increased in
High vacuum region
Ions to mass analyzer
Skimmer
To rotary pump
importance in food and nutrition science through
applications in authentication, nutrient mineral me-
tabolism and toxicology. One of the principle tech-
niques of inorganic mass spectrometry, ICP-MS, is
based on the production of ions in a high-temperature
(6000–8000 K) atmospheric-pressure argon plasma
formed by a self-sustaining electrical discharge that
is induced by a high-frequency induction coil. This
plasma is capable of ionizing most elements in the
periodic table very efficiently. Samples are introduced
into the ion source in nebulized solutions flowing at
rates of 0.1 –1 ml min1. They are then ionized effi-
ciently by the high-temperature plasma. Alternative
sample introduction techniques, including electro-
thermal vaporization, laser desorption, hydride
generation, electrospray, and combined chromatog-
raphy/mass spectrometry, are also available.
The ionization efficiency is generally very high for 0023
most nutrient and toxic elements. The plasma forms a
supersonic jet in the differentially pumped region
behind the sampling cone. Ions are introduced into
the vacuum of the mass analyzer via a series of differ-
entially pumped stages. A schematic diagram of an
ICP-MS ion source is shown in Figure 4.
Most ICP-MS instruments are based on quadru- 0024
pole mass analyzers. However, high-resolution mag-
netic sector instruments are also available. These are
capable of resolving polyatomic interferences gener-
ated in the argon plasma that may have the same mass
 MASS SPECTROMETRY/Principles and Instrumentation 3743

848.6
100

893.2

942.9

808.2

998.2

771.3
1060.5

737.7 1131.3

693.6
1211.8
706.9 1305.0

0 m/z
650 700 750 800 850 900 950 1000 1050 1100 1150 1200 1250 1300 1350 1400 1450

100
16951.8

0 Mass
16860 16880 16900 16920 16940 16960 16980 17000 17020 17040 17060 17080 17100 17120 17140

fig0003 Figure 3 Electrospray mass spectrum of horse heart myoglobin, showing the raw, multiply charged data (upper trace) and the
transformed spectrum (lower trace) showing the measured molecular weight of the sample.

as isotopes of interest, for example 40Ar 16Oþ and mass channel. This interference can be attenuated
56
Feþ. High-precision isotope ratio measurements considerably by charge exchange with H2 in the colli-
may also be conducted if multicollector magnetic sion cell.
sector mass analyzers are employed. ICP-MS is a rapid and useful multielement analysis 0026

0025 A major step forward has been the introduction of
collision cell technology to eliminate polyatomic
interference ions. These cells have been used with
quadrupole and magnetic sector instruments. The
important nutritional element selenium suffers from
technique that is also capable of determining isotope
ratios in studies of trace element metabolism in
humans.
Thermal ionization mass spectrometry (TIMS) TIMS 0027

an abundant interference peak from 40Arþ2 in the

80
Se is a well-established technique in inorganic mass
 3744 MASS SPECTROMETRY/Principles and Instrumentation
spectrometry with a much longer pedigree than ICP-
MS. It is based on the production of positive or
negative atomic or molecular ions at the surface of a
resistively heated metal filament. It has been used to
conduct stable studies of nutrient mineral metabolism
and is a ‘gold standard’ analytical technique for quan-
titative analysis of nutrients and toxicants in foods
by isotope dilution mass spectrometry, in which
measurements are conducted by adding a predeter-
mined quantity (the ‘spike’) of an enriched stable
isotope (of known isotopic composition) to the target
analyte. Isotope ratio measurement of the mineral of
interest then yields the amount of analyte precisely
and accurately.
0028 Mass analyzer The mass analyzer of the mass spec-
trometer separates ionized atoms and molecules
according to their mass-to-charge ratio, m/z. Several
different types of mass analyzer are available. The
main types of relevance to food and nutrition research
are described below.
0029 The maximum practical resolving power, R, of a
mass spectrometer is an important instrumental char-
acteristic. It is effectively a measure of the instru-
ment’s ability to separate ions. Several different
methods can be used to define R, but the two most
common are the ‘10% valley’ definition (eqn (7)),
which is based on the separation between two peaks
of equal height and mass m and m þ Dm,
m performed using other types of mass analyzer).
R¼ , ð7Þ
m þ
m
when the peaks are separated by a valley of 10% peak
height, and the full width at half height definition
(eqn (8)),
m trical potential, to yield a ‘double-focusing’ instru-
R¼ , ð8Þ

m
Distance from coil (mm)
20 15 10
Sampling
region
6200 6500 8000
Temperature K ⫾10%
fig0004 Figure 4 Schematic diagram of an ICP-MS ion source. Redrawn from a diagram supplied by VG Elemental, with permission.
where Dm is the width of a peak of mass m at 50% of
its height.
Some instruments, magnetic sector-based mass 0030
spectrometers, for example, yield constant resolution
over the entire mass range (i.e., m/Dm is constant). In
contrast, quadrupole mass analyzers yield a constant
Dm over the entire mass range. In this case, resolution
is mass-dependent, usually some multiple of the mass,
e.g., twice the mass.
Magnetic sector mass spectrometers Ions formed in 0031
the ion source region are accelerated through a po-
tential and are injected into a sector magnetic field
(See Figure 8). The motion of the ions in the magnetic
field is described by eqn (9),
B2 r2 e
m=z ¼ , ð9Þ
2V
where z is the number of charges on an ion of mass m,
e is the electronic charge, B is the magnetic field
strength, and r is the radius of the ion path. The
mass scale is generally scanned by varying, B rather
than V, as this yields the most sensitive and reprodu-
cible data. Voltage scanning is used to perform
selected ion monitoring measurements, in which
a small number of mass channels are monitored con-
secutively and repetitively. This mode is used to
record quantitative data with high sensitivity
(selected ion monitoring measurements may also be
Single-focusing magnetic sector mass spectrom- 0032
eters (now something of a rarity) are capable of a
maximum resolution of about 5000. Increased resolv-
ing power is attainable by adding an energy resolving
sector, a pair of curved plates held at a defined elec-
ment. These help to increase resolving power by
Induction coil
Sample aerosol
Body of torch
Induction
region
 MASS SPECTROMETRY/Principles and Instrumentation 3745

correcting for the spread in ion velocities that most High-resolution mass spectrometry is commonly 0034

ionization techniques produce. The electric sector can
be placed before or after the magnetic sector. Figure 5
shows a design of ‘Nier–Johnson’ geometry, in which
an electric sector deflects the ion beam through p/2
radians, before it enters a magnetic sector with a
deflection of p/3 radians.
0033 Double focusing mass spectrometers are available
in a number of designs and can attain maximum
resolving powers from 10 000 to over 200 000, de-
pending on their design.
used to measure ‘accurate masses’, i.e., masses deter-
mined to three or four decimal places. These data may
then be converted into the empirical formulae of ions
in the mass spectrum. High-resolution mass spec-
trometry may also be used to increase the specificity
of quantitative measurements, or to resolve isotopic
clusters of multiply charged peaks formed by electro-
spray so that the charge state can be measured unam-
biguously.
Isotope ratio mass spectrometers Although isotope 0035

ratios may be measured successfully using conven-

tional mass spectrometers, some applications much
require higher precision measurements than can be
2FFR attained by using scanning instruments. In such cases,
specialized high precision isotope ratio mass spec-
Magnetic
+ analyzer trometers are available. An example of this type of
Electrostatic instrument is the gas isotope ratio mass spectrometer
analyzer − (GIRMS), used to determine the isotope ratios of
gases such as CO2, N2, SO2, and H2 to a very high
degree of precision and accuracy.
The analyte is combusted or reduced to generate 0036
1FFR the gases that are ionized in a gas tight EI source. A
Ion source Collector magnetic sector separates the ion beam, with the
difference (from a conventional scanning instru-
Figure 5 Schematic diagram of double-focusing magnetic
fig0005
ment) that each mass channel is collected simultane-
sector ion optics of Nier–Johnson geometry. FFR, first field-free
region. From Chapman JR (1985) Practical Organic Mass Spec- ously, i.e., the mass spectrometer uses a multicollector
trometry, 2nd edn. Chichester, UK: Wiley-Interscience with per- ion detection system. This yields high-precision
mission. isotope ratios because any fluctuations in ion beam

Standard

Ion Magnet
beam

Combustion Ion
Sample
system Source

Faraday
collectors

Amplification
and
recording

Computer

fig0006 Figure 6 Schematic diagram of a gas isotope ratio mass spectrometer. From Mellon F, Self R and Startin JR (2000) Mass Spec-
trometry of Natural Substances in Food. London: The Royal Society of Chemistry with permission.
 3746 MASS SPECTROMETRY/Principles and Instrumentation

intensity occur simultaneously at each detector.
A diagram of a GIRMS instrument is shown in
Figure 6.
0037 The main uses of GIRMS in food and nutrition
science are in determining the authenticity of foods
traps are generally used in the scientific literature to
refer to Paul ion traps, thanks to the great success of
these instruments in compact, bench-top designs of
mass spectrometer.
An ion trap may be described as a quadrupole that 0041

and in human studies of nutrient metabolism. High-
precision multicollector mass spectrometers are also
available for use with TIMS or ICP-MS.
0038 Quadrupole mass analyzers Quadrupole mass ana-
has undergone a solid of rotation. A typical ion trap
comprises two endcap electrodes and a ring electrode,
all of hyperbolic or hemispherical cross-section
(Figure 8).
The end-cap electrodes contain small-diameter 0042

lyzers comprise four parallel rods of circular or hyper-
bolic cross-section. These are connected to radio-
frequency (V0 cos ot) and direct-current (U) power
supplies, as shown in Figure 7.
0039 The equations of motion governing the path of
ions between the rods describe a complex series
of oscillations and will not be discussed here. These
oscillations are stable for particular values of the
equations and unstable for other values. By operating
the quadrupole in a stable region, the ions will be
constrained to follow a path between the rods until
they reach the detector. The mass spectrum is scanned
holes for allowing ions to enter and leave the trap.
Ions are confined inside the trap by a radio-frequency
field of constant frequency but variable power. The
ions may be detected, according to their m/z ratio, by
applying voltages sufficient to eject them from the
trapping field. Ion traps, despite their simplicity and
cheapness of construction, can be used to perform
sophisticated tandem mass spectrometry experiments
in addition to fulfilling a role as conventional scan-
ning mass spectrometers.
FTICR mass spectrometry FTICR (sometimes 0043

by varying V0 and U so that the ratio U/V0 remains
constant. The quadrupole mass analyzer acts as a
mass filter, allowing one mass channel at a time to
reach the detector as the mass range is scanned.
Quadrupole mass spectrometers are intrinsically
simply known as Fourier transform mass spectrom-
etry) is a Penning trap in which ions confined by a
strong magnetic field, B, move in circular orbits of
characteristic frequency, o (Figure 9).
The motion of the ions is governed by the cyclotron 0044

low-resolving-power instruments but are relatively equation (eqn. (10)),

cheap and robust.
B
!¼z : ð10Þ
0040 Ion traps The Paul ion trap is related to quadrupole m
mass analyzers. The term ‘Ion Trap’ is, strictly speak- A broad-band ‘chirp’ of electromagnetic radiation 0045

ing, generic and can encompass the Penning ion traps is applied to excite all ions in the trapping cell to their
used in Fourier transform ion cyclotron resonance cyclotron frequency. The orbiting ions induce an
(FTICR) instruments (see below). However, ion alternating ‘image current’ in the walls of the cell

Detector
Filament
End cap

Ring
electrode
End cap
(U+Vcos ωt)
Electron multiplier To preamplifier
detector (Ion signal)

Amplifier and Amplifier and

Scan acquisition
RF generator, RF generator
processor
fundamental supplementary
(computer)
RF voltage RF voltage
lons in −(U + Vcos ωt)
Figure 8 Schematic diagram of an ion trap (Paul trap) mass fig0008
fig0007 Figure 7 Schematic diagram of a quadrupole mass spectrom- spectrometer. From Andrews DL (1990) Perspectives in Modern
eter. Chemical Spectroscopy. Berlin: Springer-Verlag with permission.
 MASS SPECTROMETRY/Principles and Instrumentation 3747

Receiver plate Ionization region

(pulsed EI, FAB, SIMS,
Magnetic laser or 252 Cf)
field B

Ion drift velocity (v) Recording

electronics
Transmitter Ion Transmitter and computer
plate motion plate Drift tube
(length L) Detector
Acceleration
grids

Figure 10 Schematic diagram of a ToF mass spectrometer. fig0010

From Andrews DL (1990) Perspectives in Modern Chemical Spec-

Receiver plate troscopy. Berlin: Springer-Verlag with permission.

fig0009 Figure 9 Schematic diagram of a Fourier transform ion cyclo-

tron resonance mass spectrometer. From Andrews DL (1990)
Perspectives in Modern Chemical Spectroscopy. Berlin: Springer-
Verlag with permission.
through a potential, V, and then enter a field-free drift
region (Figure 10).
According to eqn (11), the ions will all have the 0049

same kinetic energy.

receiver plates. This signal is complex because each
1
m/z value generates its own characteristic frequency. mv2 ¼ zeV, ð11Þ
A Fourier transform of this complex signal yields the 2
masses of all ions in the spectrum. Frequencies can where v is the ion velocity. The ToF, t, of the ions is
be measured with great accuracy, and so very high governed by eqn (12),
resolving powers, over 1 000 000 in some cases, are  1
attainable. However, resolution does fall off with m 2
t¼ L: ð12Þ
increasing m/z. 2zeV
0046 FTICR instruments can be used to perform se-
quential tandem mass spectrometry experiments and Early ToF instruments had limited mass resolution, 0050

are compatible with pulse ionization techniques. but ‘delayed extraction’ techniques and the introduc-
Although they are expensive to purchase, and running tion of electrostatic mirrors (reflectrons) have
costs are high, they have undergone something of increased their resolution dramatically. ToF instru-
a renaissance in recent years. This is partly due to ments are capable of a very high sensitivity because
their compatibility with newer pulse ionisation they record all the ions generated in a single ioniza-
techniques like MALDI. Perhaps more importantly, tion pulse.
the multiple charging observed in ESI of proteins
Ion detection and recording The most common 0051
yields a mass envelope in the region where full advan-
detector in mass spectrometry is the electron multi-
tage can be taken of FTICR’s high-resolution
plier; this amplifies the weak ion beam signal greatly,
performance (i.e., 600–2000 Th). This allows unam-
yielding gains of up to the order of 106. Multipliers
biguous determination of the charge state of product
can be either a discrete dynode type or a continuous
ions formed in tandem mass spectrometry experi-
dynode, generally known as a channel electron multi-
ments.
plier or channeltron. The discrete dynode electron
multiplier comprises a series of Be–Cu alloy dynodes
0047 Time of flight (ToF) mass analyzers The ToF mass
arranged as shown in Figure 11.
analyzer is now becoming widespread for several
The first dynode is held at a negative potential of 0052
reasons. First, it is compatible with MALDI, a pulsed
several kilovolts relative to the (positive ion) beam.
ionization technique that is a perfect match for ToF.
The beam strikes the first dynode and generates a
Second, improvements in design and performance
shower of secondary electrons. These electrons are
have yielded major advances in performance, particu-
then attracted to the second dynode, generating
larly resolution and sensitivity. Third, ToF mass ana-
more electrons, and so on, in a cascade through the
lyzers have, theoretically, an unlimited mass range.
device. Negative ions may be detected by using a
0048 ToF analyzers are based on the property that the
conversion dynode held at a high positive potential.
drift velocities of ions are dependent on their m/z
Channeltron detectors operate according to similar
ratio. Ions formed in the source region are accelerated
 3748 MASS SPECTROMETRY/Principles and Instrumentation
Ion beam
Electrons
fig0011 Figure 11 Schematic of a discrete dynode electron multiplier ion detector. From McFadden WH (1973) Techniques of Combined Gas
Chromatography/Mass Spectrometry. Chichester, UK: Wiley-Interscience with permission.
principles as discrete dynode electron multipliers. The
main difference is that individual dynodes are re-
placed by a tapered, coiled tube, and electrons bounce
from along the tube towards the earthed terminus.
Array detectors comprise a series of miniaturized
channeltrons that are arranged adjacent to each
other. They can be used to detect ions simultaneously
over a range of masses and are used to increase
sensitivity in some types of magnetic sector mass
spectrometer.
0053 The scintillation detector is an alternative ion beam
amplification device in quite widespread use. The ion
beam strikes a surface that emits electrons towards a
phosphor screen, generating photons. The photons
are amplified using a device akin to the image inten-
sifiers used in night-vision binoculars and similar
devices. Scintillation-based photomultiplier arrays
are also available.
0054 Faraday cup detectors are used mainly in isotope
ratio mass spectrometers. The Faraday detector is a
very simple device, comprising a conducting ‘bucket’
that collects the ion beam, yielding an electrical
signal. The minute electrical signal generated is then
amplified and recorded. Faraday cups are used in
applications where accurate measurement is more
important than sensitivity, for example in high-
precision isotope ratio measurements conducted by
GIRMS, ICP-MS, or TIMS.
0055 Tandem mass spectrometry (mass spectrometry/mass
spectrometry (MS/MS)) Tandem mass spectrom-
etry, or mass spectrometry/mass spectrometry
(MS/MS), experiments are conducted classically by
combining two mass analyzers in a single instrument.
For example, two double focusing magnetic sector or
two quadrupole mass analyzers may be combined in
series. Tandem quadrupole mass spectrometers are
generally known as ‘triple quadrupoles.’ However,
Amplifier
only two of the quadrupoles are involved in mass
analysis: the third (middle) quadrupole transmits all
ions and is used to ‘activate,’ i.e., transfer energy to,
the ions of interest.
MS/MS has been described as ‘taking the 0056
mass spectrum of an ion in a mass spectrum.’ The
principle of MS/MS is that a sample is ionized
and mass-analyzed in the first mass analyzer. A
particular m/z value of interest is selected from
the mass spectrum and is directed into a collision
cell that contains a neutral gas (argon, for example).
The ion is vibrationally excited by collision with
the target gas, a process known as collision induced
dissociation. This generates fragment ions that
are separated and recorded by the second mass
analyzer. The process is shown schematically in
Figure 12.
The example given is of a product ion scan and is 0057
used as a general technique for analyzing target com-
ponents in a mixture, or for generating characteristic
fragment ions from a sample that yields stable mo-
lecular ions under the ionization conditions used.
Other scan modes are available, including precursor
ion scans, constant neutral loss, and selected reaction
monitoring.
Tandem mass spectrometers are available in several 0058
different configurations. ‘Hybrid’ instruments, for
example magnetic sector/quadrupole or quadrupole
ToF, are also found in many laboratories. ToF mass
spectrometers may also be used in MS/MS mode. MS/
MS experiments may also be performed using ion
trap or FTICR instruments. In this case, MS/MS is
conducted by temporal, rather than spatial, separ-
ation of ions. For example, an ensemble of ions in
an ion trap is ‘swept’ using a scan function that expels
all ions from the trap except the m/z value of interest.
These ions are activated by collision with a gas
admitted to the ion trap, and the ionic products of
 MASS SPECTROMETRY/Principles and Instrumentation 3749

600 700 800 900

Mass spectrum : MS 1
Collision cell

Collision
induced
dissociation

300 500 700 800

MS/MS spectrum : MS 2

fig0012 Figure 12 Schematic representation of tandem mass spectrometry (product ion analysis).

this activation are measured by a conventional Library searching of a large database of the 0061

mass scan. This process of isolation can be repeated,
i.e., a particular product ion can, in turn, be isolated
and activated, and so on in a sequence of reactions.
This allows MS/MS/MS or MSn experiments to be
carried out.
0059 MS/MS has many analytical applications. These
include the analysis of complex mixtures without
any chromatographic separation, generation of char-
acteristic fragment ions from molecular ions, and
improving the selectivity of quantitative measure-
ments.
mass spectra of known compounds is now routine
as an aid to interpreting EI spectra and identifying
components in (for example) a complex gas chroma-
tography – mass spectrometry chromatogram. More
sophisticated, compound class-specific algorithms are
available for particular types of analysis, such as
identification of proteins in proteomics experiments.
Computer systems are now an integral component of
mass spectrometers and have greatly simplified the
process of instrument control, data acquisition, and
interpretation.

0060 Data acquisition and processing Computers, typic-

ally PCs, are now used routinely both to control
mass spectrometers and to acquire and process
mass-spectrometric data. Parameters such as focus-
ing, resolution, and scan speed may be adjusted and
set via a computer. The same computer may be used
to control mass spectrometer inlet systems such as gas
chromatography, high-performance liquid chroma-
tography, or automated direct insertion probes. The
computer is also the final link in the signal-recording
chain. It forms part of the system used to digitize
the mass spectrum, calibrate the mass scale,
enhance the quality of data, and, ultimately, to aid
in the interpretation of data. All these functions may
be carried out by a single computer coupled to a
sophisticated data digitization device.
See also: Chromatography: Gas Chromatography; Mass
Spectrometry: Applications
Further Reading
Adams F, Gijbels R and van Grieken R (1988) Inor-
ganic Mass Spectrometry. Chichester, UK: Wiley-Inter-
science.
Chapman JR (1995) Practical Organic Mass Spectrometry,
2nd edn. Chichester, UK: Wiley-Interscience.
McLafferty FW and Turecek F (1993) Interpretation of
Mass Spectra, 4th edn. Sausalito, USA: University
Science Books.
Rose ME and Johnstone RAW (1996) Mass Spectrometry
for Chemists and Biochemists, 2nd edn. Cambridge:
Cambridge University Press.