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 Mass spectrometry,
1. Also called mass spectroscopy, analytic technique by which chemical
substances are identified by the sorting of gaseous ions in electric and
magnetic fields according to their mass-to-charge ratios.
2. The instruments used in such studies are called mass spectrometers and
mass spectrographs, and they operate on the principle that moving ions
may be deflected by electric and magnetic fields.
3. The two instruments differ only in the way in which the sorted charged
particles are detected. In the mass spectrometer they are detected
electrically, in the mass spectrograph by photographic or other
nonelectrical means.
4. The term mass spectroscope is used to include both kinds of devices.
5. Since electrical detectors are now most commonly used, the field is
typically referred to as mass spectrometry.

Mass to charge ratio…

The mass-to-charge ratio (m/Q) is a physical quantity that is most widely used in the electrodynamics
of charged particles, e.g. in electron optics and ion optics.

It appears in the scientific fields of electron microscopy, cathode ray tubes, accelerator physics, nuclear
physics, Auger electron spectroscopy, cosmology and mass spectrometry.

The importance of the mass-to-charge ratio, according to classical electrodynamics, is that two particles
with the same mass-to-charge ratio move in the same path in a vacuum, when subjected to the same
electric and magnetic fields.

Its SI units are kg/C. In rare occasions the Thomson has been used as its unit in the field of mass
spectrometry.

• Common symbols: m/Q

• SI unit: kg/C

• In SI base units: kg ⋅A -1⋅s -1

Parts of mass spectroscope

• Mass spectroscopes consist of five basic parts:
 1. A high vacuum system;
2. A sample handling system, through which the sample to be investigated
can be introduced;
3. An ion source, in which a beam of charged particles characteristic of the
sample can be produced;
4. An analyzer, in which the beam can be separated into its components;
5. And a detector or receiver by means of which the separated ion beams can
be observed or collected.

Common workings of mass spectroscope

• Many investigations have been conducted with the help of mass spectrometry.
• These include ………………….
1. The identification of the isotopes of the chemical elements.
2. Determination of their precise masses and relative abundances.
3. Dating of geologic samples.
4. Analysis of inorganic and organic chemicals especially for small amounts of impurities.
5. Structural formula determination of complex organic substances.
 6. Strengths of chemical bonds and energies necessary to produce particular ions.
7. Identification of products of ion decomposition.
8. Analysis of unknown materials, such as lunar samples, for their chemical and
isotopic constituents.
9. Mass spectroscopes also are employed to separate isotopes and to measure the
abundance of concentrated isotopes when used as tracers in chemistry, biology,
and medicine.

Types of ionization methods of samples

The heart of the spectrometer is the ion source. Here molecules of the sample (black
dots) are bombarded by electrons (light blue lines) issuing from a heated filament. This
is called an EI (electron-impact) source. Gases and volatile liquid samples are allowed to
leak into the ion source from a reservoir (as shown). Non-volatile solids and liquids may
be introduced directly. Cations formed by the electron bombardment (red dots) are
pushed away by a charged repelled plate (anions are attracted to it), and accelerated
toward other electrodes, having slits through which the ions pass as a beam. Some of
these ions fragment into smaller cations and neutral fragments. A perpendicular
magnetic field deflects the ion beam in an arc whose radius is inversely proportional to
the mass of each ion. Lighter ions are deflected more than heavier ions.

Ion sources
a. Direct-current arc
b. Historically this was the first way of producing a beam of ions and came quite
naturally out of the 19th-century experiments for observing the passage of
electricity in gases at low pressure. Two planar electrodes oriented
perpendicular to the axis of the electric field can, with a few hundred-volt
potential difference, form a plasma discharge. (Plasma refers to an
ionized gas containing an approximately equal number of positive ions and
electrons.)
c. Electrons attracted to the anode collide with molecules of the gas to form ions
and free more electrons; the positive ions contribute in turn to further ionization
by their collisions. A hole in the cathode allows positive ions to emerge
collimated into a beam.
d. Such sources are found with many electrode configurations, including electron-
emitting filaments, and operate with wide ranges of pressures and voltages.
e. Sources with magnetic fields parallel to the electric fields can yield beams
greater than one milliampere.
 f. Direct-current sources were widely used during the first decades of mass
spectrometry.
g. They served well for gases and liquids introduced as vapors and for many solids
as well, because these could be transformed into gaseous atoms and
incorporated into the plasma through impact by the ions, a process
called sputtering.
h. One disadvantage of this kind of ionization is the wide band of energies
attained by the ions, ranging from the maximum electrode potential to almost
zero.
i. Such a distribution of energies was the cause of Thomson’s parabolas, but
accurate work requires a narrow energy range, which in this case must be
achieved in the analyzer section of the instrument.

Electron bombardment
1. Electrons extracted from a glowing filament may be used to ionize gases. This is the
basis for the electron bombardment ion source .A satisfactory electrode arrangement
enables the production of a beam of ions much more nearly homogeneous in energy
than with the arc, greatly simplifying the ensuing analyzing method.
2. Electron impact has remained the most widely used method of ionization in mass
spectrometry.
3. It is subject to problems common to the arc: an almost total lack of selectivity as to
the chemical element ionized and, to a lesser extent, the production of ions with
degrees of ionization greater than one.
4. Electron impact is utilized extensively in fields of study in which the sample is gaseous or
prepared in gaseous form.
5. Isotopic studies of carbon, nitrogen, oxygen, sulfur, and the noble gases make up a large
field of endeavor.
 Electron bombardment impact is useful for studying organic compounds.
Organic molecules are ionized not only as ions of the whole molecule but in a range of
fragments as well. This property, which may at first seem disadvantageous, is actually quite
valuable in organic identifications because the resulting mass spectrum allows the identification
of the source molecule as uniquely as fingerprints are used in human identification. This forms
the basis of a powerful method of organic analysis.
If the fragmentation of the molecule is harmful to the objectives of the experiment, another
method of ionization can be employed that produces few fragments. In this technique
a reagent such as methane (CH4) is mixed with the sample gas and subjected to electron
bombardment. The ionized methane (CH4/+) reacts to form CH5/+, which in turn reacts to ionize
the sample gas by proton or charge transfer. This process is called chemical ionization, and in
some cases it increases the mass of the ion formed by one unit.
HEMICAL IONIZATION METHODS IN MS
A schematic diagram of chemical ionization source

1. Chemical ionization (CI) is a soft ionization technique used in mass

spectrometry.[1][2] This was first introduced by Burnaby Munson and Frank H.
Field in 1966.[3]
2. This technique is a branch of gaseous ion-molecule chemistry.[2] Reagent gas
molecules are ionized by electron ionization, which subsequently react with
analyte molecules in the gas phase in order to achieve ionization.
3. Negative chemical ionization (NCI), charge-exchange chemical ionization
and atmospheric-pressure chemical ionization (APCI) are some of the common
variations of this technique. CI has several important applications in
identification, structure elucidation and quantitation of organic compounds.
4. Beside the applications in analytical chemistry, the usefulness in chemical
ionization extends toward biochemical, biological and medicinal fields as well.[4]
Mechanism…
1. A CI experiment involves the use of gas phase acid-base reactions in the chamber.
2. Ions are produced through the collision of the analyte with ions of a reagent gas
that are present in the ion source.
3. Some common reagent gases include: methane, ammonia, water and isobutane.
Inside the ion source, the reagent gas is present in large excess compared to the
analyte. Electrons entering the source with energy around 200-500 eV[6] will
preferentially ionize the reagent gas.
4. Then, the ion/molecule reactions produces more stable reagent ions and the
resultant collisions with other reagent gas molecules will create an
ionization plasma.
5. Positive and negative ions of the analyte are formed by reactions with this
plasma.[5]
•
Fast atom bombardment
•
(FAB) is an ionization technique used in mass spectrometry in which a beam of high
energy atoms strikes a surface to create ions
•

• It was developed by Michael Barber at the University of Manchester in 1980.

• When a beam of high energy ions is used instead of atoms (as in secondary ion mass
spectrometry), the method is known as liquid secondary ion mass spectrometry (LSIMS).
• In FAB and LSIMS, the material to be analyzed is mixed with a non-volatile chemical protection
environment, called a matrix, and is bombarded under vacuum with a high energy (4000 to
10,000 electron volts) beam of atoms.
• The atoms are typically from an inert gas such as argon or xenon. Common matrices
include glycerol, thioglycerol, 3-nitrobenzyl alcohol (3-NBA), 18-crown-6 ether, 2-
nitrophenyloctyl ether, sulfolane, diethanolamine, and triethanolamine. This technique is similar
to secondary ion mass spectrometry and plasma desorption mass spectrometry.
Ionization mechanism….
Schematic procedure of the fast atom bombardment process.

1. FAB is a relatively low fragmentation (soft) ionization technique and produces primarily intact
protonated molecules denoted as [M + H]+ and deprotonated molecules such as [M - H]−.
2. Radical cations can also be observed in a FAB spectrum in rare cases. FAB was designed as an
improved version of SIMS that allowed for the primary beam to no longer cause damaging
effects to the sample.
3. The major difference between the two techniques is the difference in the nature of the primary
beam used; ions vs atoms.
4. For LSIMS, Cesium, Cs+ ions make up the primary beam and for FAB the primary beam is made
up of Xe or Ar atoms.
5. Xe atoms are used because they tend to be more sensitive than Argon atoms due to their larger
masses and more momentum.

6. For the molecules to be ionized by FAB, first the slow moving atoms (Xe or Ar) are ionized by
colliding electrons.
7. Those slow moving atoms are then ionized and accelerated to a certain potential where they
develop into fast moving ions that become neutral in a dense cloud of excess natural gas atoms
that make a flowing stream of high translational energy atoms.
8. Although the exact mechanism of how the samples are ionized have not been fully discovered,
the nature of its ionization mechanism is similar to matrix-assisted laser desorption/ionization
(MALDI) and chemical ionization.
Matrices and sample introduction…
1. As previously stated, in FAB the samples are mixed with a non-volatile environment (matrix) in
order to be analyzed.
2. FAB uses a liquid matrix that is mixed with the sample in order to provide a sample ion current
that is sustained, reduces damages made to the sample by absorbing the impact of the primary
beam, and keeps the sample molecules form aggregating.
3. The liquid matrix, like any other matrix, most importantly provides a medium that promotes
sample ionization. The most widely accepted matrix for this type of ionization is glycerol.
4. Choosing the appropriate matrix for the sample is crucial because the matrix can also influence
the degree of fragmentation of the sample (analyte) ions.
5. The sample can then be introduced to FAB analysis.
6. The normal method of introducing the sample-matrix mixture is through an insertion probe.
7. The sample-matrix mixture is loaded on a stainless-steel sample target on the probe, which is
then placed in the ion source via a vacuum lock.
8. The alternative method of introducing the sample is by using a device called continuous flow
fast atom bombardment (CF)-FAB.

Continuous flow fast atom

bombardment…..
1. In continuous flow fast atom bombardment (CF-FAB), the sample is introduced into the mass
spectrometer insertion probe through a small diameter capillary.
2. (CF)-FAB was developed to minimize the problem of poor detection sensitivity that is caused by
an excess of the matrix background that results in a high matrix-to-sample ratio.
3. When a metal frit is used to disperse the liquid on the probe, the technique is known as frit FAB.
4. Samples can be introduced by flow injection, microdialysis, or by coupling with liquid
chromatography.
5. Flow rates are typically between 1 and 20 μL/min.[13] CF-FAB has a higher sensitivity compared
to static FAB.

Applications……
1. The first example of the practical application of this FAB was the elucidation of the amino acid
sequence of the oligopeptide efrapeptin D.
2. This contained a variety of very unusual amino acid residues.
3. The sequence was shown to be: N-acetyl-L-pip-AIB-L-pip-AIB-AIB-L-leu-beta-ala-gly-AIB-AIB-L-
pip-AIB-gly-L-leu-L-iva-AIB-X. PIP = pipecolic acid, AIB = alpha-amino-isobutyric acid, leu =
leucine, iva = isovaline, gly = glycine.
 4. This is a potent inhibitor of mitochondrial ATPase activity. Another application of FAB includes
its original use for the analysis of condensed-phase samples.
5. FAB can be use for measurements of the molecular weight of samples below 5000 Da, as well as
their structural characteristics.
6. FAB can be paired with various mass spectrometers for data analysis, such as with a quadrupole
mass analyzer, liquid chromatography–mass spectrometry, and more.

MALDI in mass spectrometry

❖
In mass spectrometry, matrix-assisted laser desorption/ionization (MALDI) is
an ionization technique that uses a laser energy absorbing matrix to create ions from
large molecules with minimal fragmentation.
❖ It has been applied to the analysis of biomolecules (biopolymers such
as DNA, proteins, peptides and carbohydrates) and various organic molecules (such
as polymers, dendrimers and other macromolecules), which tend to be fragile and fragment when
ionized by more conventional ionization methods.
❖ It is similar in character to electrospray ionization (ESI) in that both techniques are relatively soft
(low fragmentation) ways of obtaining ions of large molecules in the gas phase, though MALDI
typically produces far fewer multi-charged ions.

MALDI methodology is a three-step process.

1. First, the sample is mixed with a suitable matrix material and applied to a metal plate.
2. Second, a pulsed laser irradiates the sample, triggering ablation and desorption of the sample
and matrix material.
3. Finally, the analyte molecules are ionized by being protonated or deprotonated in the hot plume
of ablated gases, and then they can be accelerated into whichever mass spectrometer is used to
analyse them.

What is MALDI?
MALDI is the abbreviation for "Matrix Assisted Laser Desorption/Ionization."

The sample for MALDI is uniformly mixed in a large quantity of matrix.

The matrix absorbs the ultraviolet light (nitrogen laser light, wavelength 337 nm) and converts it to heat
energy. A small part of the matrix (down to 100 nm from the top outer surface of the Analyte in the
diagram) heats rapidly (in several nano seconds) and is vaporized, together with the sample.
WAVELENGTHS OF DIFFERENT LASERS AND
THEIR APPLICATIONS TO IONIZE THE
SAMPLES
What is TOF MS?
TOF MS is the abbreviation for Time of Flight Mass Spectrometry.

Charged ions of various sizes are generated on the sample slide, as shown in the diagram. A potential
difference V0 between the sample slide and ground attracts the ions in the direction shown in the
diagram.

The velocity of the attracted ions v is determined by the law of conservation of energy. As the potential
difference V0 is constant with respect to all ions, ions with smaller m/z value (lighter ions) and more
highly charged ions move faster through the drift space until they reach the detector. Consequently, the
time of ion flight differs according to the mass-to-charge ratio (m/z) value of the ion. The method of
mass spectrometry that exploits this phenomenon is called Time of Flight Mass Spectrometry.
Ion separation methods
[by using different kinds of ion filters]
The separation of ions according to their mass is accomplished with static magnetic fields, time-
varying electric fields, or methods that clock the speeds of ions having the same energies—the
time-of-flight method. Static electric fields cannot separate ions by their mass but do separate
them by their energy and so provide an important design element by functioning as an energy
filter; they are described here along with magnetic fields.

Magnetic field analysis

1. Ions of mass m and charge z moving in vacuom with a velocity v in a direction
perpendicular to a magnetic field B will follow a circular path with radius r given by

2.
3. Therefore, all ions with the same charge and momentum entering the magnetic field
from a common point will move in the same radius r and will come to a first-order
focus after 180°, as shown in, regardless of their masses.
4. Hence, the mass spectrometer used by Dempster can be referred to as a “momentum
spectrometer.” If all ions of charge z enter the magnetic field with an identical kinetic
energy zV, owing to their acceleration through a voltage drop V, a definite
velocity v will be associated with each mass, and the radius will depend on the mass.
Since zV = 1/2mv 2, substitution in the previous equation will give m/z = B2r2/2V.
5. This formula shows that the radius of curvature r for ions in this spectrometer
depends only on the ratio of the ions’ mass to charge, as long as their kinetic energy is
the same.
6. Thus, a magnetic field can be used to separate a monoenergetic ion beam into its
various mass components.
Electrostatic field analysis

1. An electrostatic field that attracts ions toward a common centre—i.e., a radial field—
will also exert a focusing action on a divergent beam of ions as shown in Figure 3.
2. The radial force on the ions due to the electrostatic field will be Ez, the product of the
field E and the ionic charge z, and is equal to the centripetal force mv2r, of
mass m moving with velocity v about a radius r. Thus, one may write the equation

Combined electric and magnetic field

analysis

1. Use of an electric wedge or sector to obtain a monoenergetic beam of ions, which is

then separated for mass analysis by a magnetic sector, is another possible technique.
 2. General equations for combined electric and magnetic sectors have been developed;
they show that a suitable combination of fields will give direction focusing for
an ion beam of given mass-to-charge ratio, even though the beam may
be heterogeneous in energy.
3. The term double focusing is used for those combinations in which the angular and
velocity aberrations effectively cancel.

Time of flight

1. The simplest form of mass analysis that does not use magnetic fields depends on the
differing speeds of ions with the same energy but different masses.
2. The ion source is generally of the electron-impact type and has one or more electrodes
modulated so as to extract ions for a time that is brief compared with the time it takes
them to reach the detector.
3. The ion velocities, as given above by v = Square root of√2zV/m, and the distance
between source and detector allow the mass to be calculated directly.
4. In practice, the response of the detector is displayed on a cathode-ray oscillograph
and recorded by a computer.
5. This method has two advantages: it is fast and, if desired, can display the entire mass
spectrum. Its deficiencies are poor resolution, poor accuracy of signal, and
poor efficiency, due to the short period during which ions are extracted from the
source.
Ion-trap methods

1. It is possible to configure electric and magnetic fields so that ions can be held in stable
orbits for a period of time long enough to perform useful measurements on them.
2. Two forms of mass spectrometers are derived from this idea, the omegatron and
the Fourier-transform spectrometer.
3. Both make use of the cyclotron principle (see particle accelerator: Cyclotrons), in
which positive ions produced by a beam of electrons flowing along the axis of a
uniform magnetic field follow circular trajectories with a radius proportional to
momentum, r = mv/zB, and a frequency of rotation inversely proportional to mass, ω
= v/r = zB/m. In the omegatron the frequency of an oscillator is varied so as to bring
ions of various masses in tune and by so doing increase their momenta until they
reach a radius at which a detector is located.
4. Mass can be directly calculated from frequency. Resolution can be remarkably high if a
sufficient magnetic field is provided, but this analyzer is most frequently operated
with less than ideal resolution as a device for analyzing the residual gas of a vacuum,
information that can be extremely valuable in diagnosing the problems that often
befall such systems

Tandem spectrometry
1. The combination of two analytical techniques, such as resulted in the gas
chromatograph–mass spectrometer, has been followed by the combination of two
mass spectrometers, which has proved helpful in determining the structure of
complicated molecules.
2. A beam from the first spectrometer is passed into a gas cell (maintained in the vacuum
system by differential pumping), where it is dissociated by collisions, and these
 fragments are passed on to a second mass analyzer, which generally discloses a more
easily identified spectrum.

Quadrupole spectrometer

1. Positive ions incident along an axis parallel to four cylindrical electrodes, experience
for the static potentials indicated a focusing force along the x axis and a defocusing
one in the z direction.
2. If one superimposes a radio frequency voltage onto the static voltage, oscillatory ion
trajectories can be found that allow ions of a given mass to pass through the
quadrupole with other masses being defocused and lost from the beam.
3. A knowledge of the potentials and frequency specifies the mass.
4. This device is widely used where speed of data acquisition and high transmission are
important.
5. It is compact and lightweight and can trade off sensitivity for resolution by simple
adjustment of electrical parameters. It is the common analyzing element of gas
chromatograph–mass spectrometers.
 IMPORTANCE
Mass spectrometry has gradually become an important tool in food quality and safety evaluation,
playing an important role in the analysis of food quality components, pesticide residues,
environmental persistent pollutants, and others in recent years, thus ensuring the quality,
reliability, and safety of food.

References
https://en.wikipedia.org/wiki/Fast_atom_bombardment
https://www.shimadzu.com/an/products/maldi/ms-applications/principle-of-
malditofms/index.html
https://en.wikipedia.org/wiki/Matrix-assisted_laser_desorption/ionization
https://www.britannica.com/science/mass-spectrometry/Negative-ions

researcher- Muhammad Talha

institute-KFUEIT.PK
email-ak9651651@gmail.com
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Muhammad TALHA | Student | Khwaja Fareed University of Engineering & Information Technology,
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