Chapter Five

Analytical Mass Spectrometry

 Overview
• Principle
• Mass Analyzers
• Atomic Mass Spectrometer:
Instrumentation
Ion Sources
Mass Analyzers
Types
Application
• Molecular Mass Spectrometer
Instrumentation
Ion Sources
Mass Analyzers
Application
 Overview

Mass spectrometry (MS) is a powerful and versatile analytical tool for

obtaining information about the identity of an unknown compound, its
molecular mass, its elemental composition, and also chemical structure.
Mass spectrometry can be conveniently divided into atomic or elemental
mass spectrometry and molecular mass spectrometry.

 Atomic mass spectrometry is a quantitative tool that can determine

nearly all the elements in the periodic table. Detection limits are often
several orders of magnitude better than optical methods.

 Molecular mass spectrometry is capable of providing information about

the structures of inorganic, organic, and biological molecules and about
the qualitative and quantitative composition of complex mixtures.
 Basic Principle

A mass spectrometer generates multiple ions from

the sample under investigation, it then separates
them according to their specific mass-to-charge
ratio (m/z), and then records the relative
abundance of each ion type.
 Mass-to-Charge (m/z) Ratio

The mass-to-charge ratio, m/z, of an ion is the quantity of most interest because
the mass spectrometer separates ions according to this ratio.

The mass-to-charge ratio of an ion is the unit less ratio of its mass number to the
number of fundamental charges z on the ion.

Example, 12C1H + m/z = 16.032/1 = 16.032

4 ,
13C1H 2+, m/z = 17.032/2 = 8.516.
4
.
 The first step in the mass spectrometric analysis of compounds is the production of
gas phase ions of the compound, basically by electron ionization.
 This molecular ion undergoes fragmentation. Each primary product ion derived
from the molecular ion, in turn, undergoes fragmentation, and so on.
 The ions are accelerated into the mass analyzer, which then separated in the mass
spectrometer according to their mass-to-charge ratio, and are detected in proportion
to their abundance.
 The ions of particular m/z values are then collected and converted into an electrical
signal by the ion transducer.
 A mass spectrum of the molecule is thus produced. It displays the result in the form of
a plot of ion abundance versus mass-to-charge ratio. It may also include
comparison to known spectra, tabulation of results, and data storage.
 Ions provide information concerning the nature and the structure of their precursor
molecule.
 In the spectrum of a pure compound, the molecular ion, appears at the highest value
of m/z (followed by ions containing heavier isotopes) and gives the molecular mass
of the compound.
 Analysis by GC-MS

GC MS

Mixture Separation Identification

B
m/z

A
m/z

C
m/z
A+ B+ C B A C
 Mass spectrum

A mass spectrum is a
plot of ion abundance
versus mass-to-charge
ratio or just mass for
singly charged ions.
 Components of Mass Spectrometer

Mass spectrometers require an elaborate vacuum system to maintain a low

pressure in all of the components except the signal processor and display.

Low pressure ensures a relatively low collision frequency between various species
in the mass spectrometer that is vital for the production and maintenance of free
ions and electrons.
• Atomic mass spectrometry: The ionization source is outside the
evacuated region and also serves as the inlet. In atomic mass
spectrometers, ionization is accomplished by applying thermal or electrical
energy. The output of the ion source is a stream of positive (most common)
or negative gaseous ions.

• Molecular mass spectrometry: Samples enter the evacuated region of the

mass spectrometer through the inlet system. Solids, liquids, and gases
may be introduced depending on the nature of the ionization source. The
purpose of the inlet system is to introduce a micro amount of sample into
the ion source where the components of the sample are converted into
gaseous ions by bombardment with electrons, photons, ions, or molecules.
 Mass Spectrometer Block Diagram

Turbo
High Vacuum System molecular
pumps

Ion Mass Data

Inlet source Analyzer Detector System
 Sample Introduction

High Vacuum System

Inlet Ion Mass Data

Source Analyzer Detector System

HPLC
Flow injection
Sample plate
 Ion Source

High Vacuum System

Inlet Ion Mass Data

Source Analyzer Detector System

MALDI
ESI
FAB
LSIMS
EI
CI
 Mass Analyzer

High Vacuum System

Ion Mass Data

Inlet source Analyzer Detector System

Time of flight (TOF)

Quadrupole
Ion Trap
Magnetic Sector
FTMS
 Detector

High Vacuum System

Ion Mass Data

Inlet source Analyzer Detector System

Microchannel Plate
Electron Multiplier
Hybrid with photomultiplier
 Data System

High Vacuum System

Ion Mass Data

Inlet source Analyzer Detector System

PC
Sun SPARK Station
DEC Station
Mass Analyzer
In the magnetic sector analyzer,
shown in Figure 29-3, separation is
based on the deflection of ions in a
magnetic field.

The trajectories that ions take depend

on their m/z values. Typically, the
magnetic field is slowly changed to
bring ions of different m/z value to a
detector.

In the double-focusing mass

spectrometer, an electric sector
precedes the magnetic sector. The Figure 29-3 Schematic of a 10-7 torr magnetic sector
electrostatic field serves to focus a spectrometer. The kinetic energy, KE, of an ion of mass m
beam of ions having only a narrow and charge z exiting slit B is KE= zeV = ½ mv2. If all ions
range of kinetic energies onto a slit have the same kinetic energy, heavier ions travel at lower
velocities than lighter ions. The balancing of centripetal
that leads to the magnetic sector. force and magnetic force results in ions of different mass
Such instruments are capable of traveling different paths as shown.
very high resolution.
 Quadrupole Mass Analyzers
• The quadrupole mass analyzer consists of four cylindrical rods, as
illustrated in Figure 29-4.

• Quadrupole analyzers are mass filters that only allow ions of a certain
mass-to-charge ratio to pass.

• Ion motion in electric fields is the basis of separation. Rods opposite

each other are connected to dc and radio-frequency (RF) voltages.
With proper adjustment of the voltages, a stable path is created for ions
of a certain m/z ratio to pass through the analyzer to the transducer.

• The mass spectrum is obtained by scanning the voltages applied to the

rods.

• Quadrupole analyzers have relatively high throughput but relatively

low resolution. Unit mass (1Da) is the typical resolution of a
quadrupole analyzer.
Quadrupole mass spectrometers

Figure 29-4 A quadrupole mass analyzer

The quadrupole instrument

Vacuum

DC and AC Voltage
 Time-of-Flight Mass Analyzers
The time-of-flight (TOF) mass spectrometer represents another approach to mass
analysis. In a TOF analyzer, a packet of ions with nearly identical kinetic energies is
rapidly sampled, and the ions enter a field-free region. Since the kinetic energy, KE,
is ½mv2, the ion velocity v varies inversely with its mass, as shown by Equation 29-2:

The time required for the ions to travel a fixed distance to the detector is thus
inversely related to the ion mass.

In other words, ions with low m/z arrive at the detector more rapidly than those
with high m/z. Each m/z value is then detected in sequence. Flight times are quite
brief, leading to analysis times that are typically on the order of microseconds.

Time-of-flight instruments are relatively simple and rugged and have nearly
unlimited mass range. The TOF analyzer suffers, however, from limited resolution
and sensitivity. As a result, TOF analyzers are less widely used than magnetic
sector and quadrupole analyzers.
 Time-of-flight (TOF) Mass Analyzer
Source Drift region (flight tube)

+
+

detector
+ +

• Ions are formed in pulses.

• The drift region is field free.
• Measures the time for ions to reach the detector.
• Small ions reach the detector before large ones.
Time-of-flight (TOF) Mass Spectrometer
 Ion trap mass spectrometer
• The ion trap uses three electrodes to trap ions in small volumes.
• Various voltages are applied to the ring electrodes as well as to the entrance
and exit endcap electrodes. A cavity is created were the ions are trapped.
• Depending on different voltage settings, ions at a specific m/z is ejected
and detected.
Ion Trap Mass Analyzer

Top View

Cut away side view

 Transducers for Mass Spectrometry

Figure 29-5 Discrete-dynode electron multiplier. Dynodes are kept at

successively higher voltages by means of a multistage voltage divider.
The most common transducer is the electron multiplier, illustrated in Figure 29-5.
The discretedynode electron multiplier operates much like the photomultiplier
transducer for UV/visible radiation, discussed in Section 25A-4. When energetic
ions or electrons strike a Cu-Be cathode, secondary electrons are emitted. These
electrons are attracted to dynodes that are each held at a successively higher
positive voltage. Electron multipliers with up to 20 dynodes are available. These
devices can multiply the signal strength by a factor of up to 107.

Continuous-dynode electron multipliers are also popular. These multipliers are

trumpet-shaped devices made of glass heavily doped with lead. A potential of 1.8
to 2 kV is imposed across the length of the device. Ions that strike the surface eject
electrons that skip along the inner surface ejecting more electrons with each
impact.

In addition to electron multiplier transducers, Faraday cup transducers and array

transducers have become available for mass spectrometry. As in optical
spectrometry, array transducers allow the simultaneous detection of multiple
resolution elements. Microchannel plate arrays and micro-Faraday arrays have
been used.
 Atomic Mass Spectrometry

 Atomic mass spectrometry has been around for many years, but the
introduction of the inductively coupled plasma (ICP) in the 1970s and
its subsequent development for mass spectrometry led to the successful
commercialization of ICPMS by several instrument companies. Today,
ICPMS is a widely used technique for the simultaneous determination
of over 70 elements in a few minutes.

 The ion source is the major difference between atomic and molecular
mass spectrometry. For atomic mass spectrometry, the ion source must
be very energetic to convert the sample into simple gas phase ions and
atoms. In molecular mass spectrometry, the ion source is much less
energetic and converts the sample into molecular ions and fragment
ions.
 Sources for Atomic Mass Spectrometry
Several different ionization sources have been proposed for atomic mass
spectrometry. Table 29-2 lists the most common ion sources and the typical
mass analyzers used with each.
 The Inductively Coupled Plasma

The inductively coupled plasma is described extensively in Section 28B-2 in

connection with its use in atomic emission spectrometry. The axial geometry shown
in Figure 28-7 is most often used in ICPMS.

In MS applications, the ICP serves as both anatomizer and an ionizer.

Solution samples may be introduced by a conventional or an ultrasonic nebulizer.

Solid samples can be dissolved in solution or volatized by means of a high-voltage
spark or high-powered laser prior to introduction into the ICP.

Ions formed in the plasma are then introduced into the mass analyzer, often a
quadrupole, where they are sorted according to mass-to-charge ratio and detected.
Extracting ions from the plasma can present a major technical problem in
ICPMS. While an ICP operates at atmospheric pressure, a mass spectrometer
operates at high vacuum, typically less than 1026 torr. The interface region
between the ICP and the mass spectrometer is thus critical to ensure that a
substantial fraction of the ions produced are transported into the mass analyzer.

The interface usually consists of two metal cones, called the sampler and the
skimmer.Each cone has a small orifice (< 1 mm) to allow the ions to pass
through to ion optics that then guide them into the mass analyzer.

The beam introduced into the mass spectrometer has about the same ionic
composition as the plasma region from which the ions are extracted.
ICPMS spectra are often remarkably simple compared with conventional ICP
atomic emission spectra. The ICPMS spectrum shown in the figure consists
of a simple series of isotope peaks for each element present along with some
background ionic peaks. Background ions include Ar+1, ArO+, ArH+, H2O+,
O+, O2+, and Ar2+, as well as argon adducts with metals.

In addition, some polyatomic ions from constituents in the sample are also
found in ICP mass spectra. Commercial instruments for ICPMS have been on
the market since 1983. ICPMS spectra are used for identifying the elements
present in the sample and for determining these elements quantitatively.
Usually, quantitative analyses are based on calibration curves in which the
ratio of the ion signal for the analyte to that for an internal standard is plotted
as a function of concentration.
Schematic diagram of ICPMS
Figure 29-6 Comparison of ICP atomic emission spectrum for 100 ppm cerium (a)
with ICP mass spectrum for 10 ppm cerium (b). (Adapted from M. Selby and G. M.
Hieftje, Amer. Lab., 1987, 19, 16.)
Other Ionization Sources of Atomic Mass Spectrometry

Of the sources listed in Table 29-2, the spark source and the glow
discharge have received the most attention.

Spark source atomic mass spectrometry (SSMS) was first introduced in

the 1930s as a general tool for multielement and isotope trace analyses.
It was not until 1958, however, that the first commercial spark source
mass spectrometer appeared on the market. After a period of rapid
development in the 1960s, the use of this technique leveled off and then
declined with the appearance of ICPMS.

Currently, spark source mass spectrometry is still applied to solid

samples that are not easily dissolved and analyzed by ICP. In addition,
spark sources are used in conjunction with ICP sources to volatilize and
atomize solid samples before introduction to the plasma.
As discussed in Section 28B-5, the glow-discharge source is a useful device
for various types of atomic spectroscopy.
In addition to atomizing samples, it also produces a cloud of positive analyte
ions from solid samples. This device consists of a simple two electrode
closed system containing argon at a pressure of 0.1 to 10 torr. A voltage of 5
to 15 kV from a pulsed dc power supply is applied between the electrodes,
causing the formation of positive argon ions, which are then accelerated
toward the cathode.
The cathode is fabricated from the sample, or the sample is deposited on an
inert metal cathode. Just as in the hollow-cathode lamp (see Section 28D-2),
atoms of the sample are sputtered from the cathode into the region between
the two electrodes, where they are converted to positive ions by collision
with electrons or positive argon ions. Analyte ions are then drawn into the
mass spectrometer by differential pumping.
The ions are then filtered in a quadrupole analyzer or dispersed with a
magnetic sector analyzer for detection and determination. Glow-discharge
sources, like spark sources, are often used with ICP torches. The glow
discharge serves as the atomizer, and the ICP torch is the ionizer.
 Atomic Mass Spectra and Interferences
Interference effects in atomic mass spectroscopy fall into two broad categories:
spectroscopic interferences and matrix interferences.

Spectroscopic interferences occur when an ionic species in the plasma has the
same m/z value as an analyte ion. Most of these interferences are from
polyatomic ions, elements having isotopes with essentially the same mass,
doubly charged ions, and refractory oxide ions. High-resolution spectrometers
can reduce or eliminate many of these interferences.

Matrix effects become noticeable when the concentrations of matrix species

exceed about 500 to 1000 μg/mL. Usually, these effects cause a reduction in the
analyte signal, although enhancements are sometimes observed. Generally, such
effects can be minimized by diluting the sample, by altering the introduction
procedure, or by separating the interfering species. The effects can also be
minimized by the use of an appropriate internal standard, an element that has
about the same mass and ionization potential as the analyte.
 Detection Limits of Atomic Mass Spectrometry
• Most elements can be detected well below the part per billion level.
Quadrupole instruments typically allow ppb detection for their entire mass
range. High-resolution instruments can routinely achieve sub-part-per-
trillion detection limits because the background levels in these
instruments are extremely low.
• Quantitative analysis is normally performed by preparing calibration
curves using external standards. To compensate for instrument drifts,
instabilities, and matrix effects, an internal standard can be added to the
standards and to the sample. Multiple internal standards are sometimes
used to optimize matching the characteristics of the standard to that of
various analytes.
• For simple solutions where the composition is known or the matrix can be
matched well between samples and standards, accuracies can be better than
2% for analytes at concentrations 50 times the detection limit.
• For solutions of unknown composition, accuracies of 5% are typical.
Applications of Atomic Mass Spectrometry

ICPMS is well suited for multi element analysis and for

determinations such as isotope ratios.

ICPMS is finding widespread use in the semiconductor and

electronics industry, in geochemistry, in environmental analyses, in
biological and medical research, and in many other areas.
 Molecular Mass Spectrometry
Molecular mass spectrometry was first used for routine
chemical analysis in the early 1940s when the petroleum
industry adopted the technique for quantitative analysis of
hydrocarbon mixtures produced in catalytic crackers.

Beginning in the 1950s, commercial instruments began to be

adapted by chemists for the identification and structural
elucidation of a wide variety of organic compounds.

This use of the mass spectrometer combined with the

invention of nuclear magnetic resonance and the
development of infrared spectrometry revolutionized the
way organic chemists identify and determine the structure of
molecules.
 Figure 29-7 Mass spectrum of ethyl benzene.

To obtain this spectrum, ethyl benzene vapor was bombarded with a stream of
electrons that led to the loss of an electron by the analyte and formation of the
molecular ion M1, as shown by the reaction

The charged species C6H5CH2H.+ is the molecular ion. As indicated by the dot, the
molecular ion is a radical ion that has the same molecular mass as the molecule.
The collision between energetic electrons and analyte molecules usually imparts
enough energy to the molecules to leave them in an excited state. Relaxation then
often occurs by fragmentation of part of the molecular ions to produce ions of
lower masses.

For example, a major product in the case of ethyl benzene is C6H5CH2+, which
results from the loss of a CH3 group.

Other smaller positively charged fragments are also formed in lesser amounts.

The positive ions produced on electron impact are attracted through the slit of a
mass spectrometer, where they are sorted according to their mass-to-charge ratios
and displayed in the bar graph form of a mass spectrum.

The largest peak at m/z = 91, termed the base peak, has been arbitrarily assigned
a value of 100. The heights of the remaining peaks are then computed as a
percentage of the base-peak height.
The starting point for a mass spectrometric analysis is the formation of gaseous
analyte ions, and the scope and the utility of a mass spectrometric method is
dictated by the ionization process. The appearance of mass spectra for a given
molecular species is highly dependent on the method used for ion formation.

Table 29-3 lists many of the ion sources that have been used in molecular mass
spectrometry. These methods fall into two major categories: gas-phase sources
and desorption sources. With a gas-phase source, the sample is first vaporized
and then ionized.

With a desorption source, the sample in a solid or liquid state is converted

directly into gaseous ions. An advantage of desorption sources is that they are
applicable to nonvolatile and thermally unstable samples.
The most widely used source is the electron impact (EI) source. In this source,
molecules are bombarded with a high-energy beam of electrons. This produces
positive ions, negative ions, and neutral species. The positive ions are directed
toward the analyzer by electrostatic repulsion.

In EI, the electron beam is so energetic that many fragments are produced. These
fragments are very useful in identifying the molecular species entering the mass
spectrometer. Mass spectra for many libraries of MS data have been collected using
EI sources.

A wide assortment of ambient MS techniques exists, but desorption electrospray

ionization (DESI) and direct analysis in real time (DART) are the leading
techniques.
In addition, low-temperature plasma probe ionization (LTP), easy ambient
sonic-spray ionization (EASI), and laser ablation electrospray ionization
(LAESI) have shown promise.
 Molecular Mass Spectrometric Instrumentation

Inlet System:
The purpose of the inlet system is to introduce a representative sample to the ion
source with minimal loss of vacuum. Most modern mass spectrometers are
equipped with several types of inlets to accommodate various kinds of samples.
The major types of inlets can be classified as batch inlets, direct probe inlets,
chromatographic inlets, and electrophoretic inlets.

The conventional inlet system is the batch type in which the sample is volatilized
externally and then allowed to leak into the evacuated ionization region.

Liquids and gases can also be introduced in this way. Solids can be placed on the
tip of a probe, inserted into the vacuum chamber, and evaporated or sublimed by
heating.

Nonvolatile liquids can be introduced through special controlled-flow inlets, or

they can be desorbed from a surface on which they are coated as a thin film.
 In general, samples for molecular mass spectrometry must be pure because the
fragmentation that occurs causes the mass spectrum of mixtures to be difficult to
interpret. Gas chromatography is an ideal way to introduce mixtures because the
components are separated from the mixture by the chromatograph prior to
introduction to the mass spectrometer. The combination of gas chromatography
and mass spectrometry is often called GC/MS.

Figure 29-8 Schematic of a typical capillary GC/MS instrument. The effluent from the
GC is passed into the inlet of the mass spectrometer, where the molecules in the gas are
ionized and fragmented, analyzed, and detected.
Figure 29-9 Block diagram of a tandem mass spectrometer.
 Mass Analyzers
All of the mass analyzers listed in Table 29-1 are used in molecular mass spectrometry.
The quadrupole mass analyzer is commonly used with GC/MS systems. Higher
resolution spectrometers (magnetic sector, double-focusing, time-of-flight, Fourier
transform) are often used when fragmentation patterns are to be analyzed for structural
or identification purposes.
Tandem mass spectrometry, also called mass spectrometry-mass
spectrometry (MS/MS), is a technique that allows the mass spectrum of a
preselected or fragmented ion to be obtained.

Figure 29-9 illustrates the basic concept. With a tandem mass

spectrometer, an ionization source produces molecular ions and fragment
ions. These are then the input to the first mass analyzer, which selects a
particular ion (the precursor ion) and sends it to the interaction cell.

In the interaction cell, the precursor ion can decompose spontaneously,

react with a collision gas, or interact with an intense laser beam to produce
fragments, or product ions. These ions are then mass analyzed by the
second mass analyzer and detected by the ion detector.
Tandem mass spectrometers can produce a variety of different spectra. Production
spectra are obtained by scanning mass analyzer while mass analyzer is held
constant to act as a mass selector for the precursor ion.

A precursor-ion spectrum can be obtained by scanning mass analyzer and

selecting a given product ion with mass analyzer. If both mass analyzers are
scanned with a small offset in mass between them, a neutral loss spectrum can be
obtained. A neutral loss spectrum might be used, for example, to identify the m/z
values of all ions losing a common molecule, such as water. Finally, a complete
three-dimensional MS/MS spectrum can be acquired by recording a product ion
spectrum for each selected precursor ion, that is, by scanning mass analyzer 2 for
various settings of mass analyzer.

Tandem mass spectrometry can produce an enormous amount of information and

has proven useful in structural elucidation as well as in the analysis of mixtures.
Conventional mass spectrometry of mixtures usually requires chromatographic or
electrophoretic separation to present a single compound at a time to the mass
spectrometer.
 Ionization Techniques
• The ionization takes place in the ion source.
• Samples from GC interface are in gas-phase when introduced into the MS.
The MS is operated in vacuum to make sure analytes are vaporized and to
avoid collisions between analytes and other compounds.
• Two different ionization methods
• Electron Impact Ionization (EI)
• Chemical Ionization (CI)

Computer
system

Ionization of Separation of
Sample ions by mass Detector
introduction sample in the
ion source analyzer
Ion Sources make ions from sample molecules (Ions are easier to detect than
neutral molecules.)

Sample Inlet Nozzle Partial

Pressure = 1 atm vacuum
Inner tube diam. = 100 um (Lower Voltage)

N2 MH+
+
++++ + ++ + +
++ + +
++
++ + +
Sample in solution
++
+
+ ++ +
+
++
+ +
+ + MH2+
++ ++ + ++ +
N2 gas ++ + +

MH3+

High voltage applied Charged droplets

to metal sheath (~4 kV)

Electrospray ionization
 Electron Impact Ionization (EI)
• Most widely used method
• Analytes are bombared with high-energy electrons (usually 70eV)
• As a result of collision, an electron is removed from the analytes (M), generating
a molecular ion M+ (radical cation)
M + e- M+ + 2e-
 Electron Impact Ionization (EI)
• Due to excess internal energy, fragmentation of the molecular ion will occur.
• The fragmentation is reproducible and characteristic of the compund.
• It is also possible to predict the fragmentation on the basis of chemical
structures which is why MS is good tool for structure elucidation of unknown
compounds
 Drawbacks with EI

• Sometimes the fragmentation is too great depending on

the stability of the sample molecules and the molecular
ion will not show up in the mass spectra.
• It is then possible to reduce the ionization voltage, but
the disadvantage is that the fragmentation pattern will
change and the obtained spectra cannot be compared to
”standard” litterature spectra.
• Another solution is to use chemical ionization (CI)!

Watson, Introduction to Mass Spectrometry, 4th ed

 Chemical Ionization (CI)

• Softer ionization technique Less fragmentation  Easier to find

molecular ions.

• Two different modes: Negative Chemical Ionization (NCI) and Positive

Chemical Ionization (PCI).

• NCI is used for analytes that are able to form stable negative ions, for
example samples containing acidic groups or halogens. NCI is often used
to analyze pesticides (contains Cl or Br).

• PCI is used for samples that can form positive ions (most compounds).
 Chemical Ionization (CI)
• The principle for NCI and PCI is similar:

• Reagent gas (usually methane, isobutane or ammonia) is introduced into the

source where it is ionized:
PCI (simplified): CH4 + e- CH4+ +2e-
CH4 + CH4+ CH3 + C H5+
NCI: CH4 + e- CH4-

• The ionized gas collide with the sample molecules generating a [M+H]+ or [M+H]-
ion that is detected:

PCI: CH5+ + M  [M+H]+ + C H4

NCI: CH4- + M  [M+H]- + C H4
 EI spectra/PCI Spectra

Harris. Quantitative Chemical Analysis, 6th ed.

Matrix Assisted Laser Desorption Ionization (MALDI)

Sample plate
Laser
hn
1. Sample is mixed with matrix (X) and
dried on plate.
2. Laser flash ionizes matrix
MH+ molecules.
3. Sample molecules (M) are ionized
by proton transfer:
XH+ + M  MH+ + X.

+/- 20 kV Grid (0 V)
 The mass spectrum shows the results

40000 MH+
Relative Abundance

30000

(M+2H)2+
20000

10000
(M+3H)3+

50000 100000 150000 200000

Mass (m/z)
MALDI TOF spectrum of IgG
 Applications of Molecular Mass Spectrometry
Identification of Pure Compounds, Analysis of Mixtures, Quantitative Determinations