National Polytechnic Institute

Higher School of Chemical Engineering and Extractive Industries

Department of Industrial Chemical Engineering

Academy of Analytical Chemistry
Specialization in Instrumental Analysis

Advanced Instrumental Techniques Laboratory

Practice No. 1

IDENTIFICATION OF COMPONENTS OF A MASS SPECTROMETER

Team: No. 4

Section: B

Members:

Anaya Flores Diana Laura

Ramírez Anaya Adriel
Sánchez Pino Javier Jefté

Group:4IM86

Professor:

M and C. Karla Jenny Lozano Rojas

Fecha de Entrega:21 /Febrero /2019

OBJECTIVE:

 The student will identify the main parts of a mass spectrometer, as well as
recognize them physically.
 The student will assemble and disassemble a mass spectrometer properly.

THEORETICAL FOUNDATION:

Timeline of Mass Spectrometry

1912: Josep John Thomson creates the first mass spectrometer as he was interested in
measure the mass-charge ratio of the electron.

Figure No. 1 "Josep John Thomson"

1918: Arthur Jeffrey Dempster discovers electro-ionization as well as the magnetic approach
due to his research on the existence of the uranium-235 isotope.

Figure No. 2 'Arthur Jeffrey Dempster'

1946: William E. Stephens creates the TOF (time of flight) for mass spectrometry technique.
flight by its acronym in English) in response to your research on nuclear energies.

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 Figure N°3 'William E. Stephens'

1952: Alfred Otto Carl Nier was the first to use mass spectrometry to isolate
uranium-235, which was used to demonstrate that 235 U could undergo fission and developed the
configuration of the mass spectrometer of the sector now known as Nier geometry
Johnson consists of an electrical sector of 90°, an intermediate drift length
long and a magnetic sector of 60° in the same direction of curvature.

Figure No. 4 "Alfred Otto Carl Nier"

1953-1958: Paul, Steinwedel, Raether, Reinhard, and von Zahn create the quadrupole analyzer for
the EM technique which consists of achieving that ions with a certain charge-to-mass ratio
describe stable trajectories in a hyperbolic electrostatic field, which is achieved
applying constant and alternating potentials to four conductor bars.

Figure No. 5 'Quadrupole System'

1955: W. C. Wiley and I. H. McLaren created an advanced design of the time of flight.
by Stephen.

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 Figure No. 6 "Modified TOF Diagram"

1956: Fred Warren McLafferty and Roland Gohlke were the first to combine the technique of
gas chromatography with mass spectrometry (GC-MS).

Figure No. 7 "GC-MS System"

1956: Beynon improves the already established technique and sends it to a high resolution one which has

greater precision as well as better accuracy.

Figure No. 8 "High-resolution masses"

1966: Munson and Field achieve chemical ionization due to the electronics appearing the ion
molecular to study.

Figure No. 9 "Chemical Ionization Mechanism"

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1968: Researchers Dole, Mack, Hines, M. Ferguson, and Alice succeeded in achieving ionization by
electrospray (ESI).

Figure No. 10 "Ionization by electrospray"

1987-1988: Researchers Karas, Bachmann, Bahr, Hillenkamp, Tanaka, Waki, Ido, Akita and
Yoshida manages to couple the TOF with the MALDI ionizer (Matrix-Assisted Laser Desorption/Ionization).

which consists of a gentle ionization of the analyte that causes the vaporization of intact
thermolabile, non-volatile molecules such as proteins and lipids in a weight range
molecular between 2 to 20 kDa, at a relatively low cost and immediate result, the utility of this
the technique is due to obtaining a specific spectrum of an organism and comparing it with
databases, identify at the level of gender, species and even isolated bacterial strains and
fungal.

Figure No. 11 'MALDI'

1989: Henry, Williams, Wang, Mc. Lafferty, Shabanowits and Hunt create for the first time the

mass spectrometry using electrospray ionization as well as ion resonance

cyclotron due to the research of crude oil as this technique achieves high
accuracy and clarity of the compounds present in the mixtures of said fuel.

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 Figure No. 12 'ionization by electrospray implemented in a mass spectrometer'

1990: Researchers Katta and Chait manage to observe conformational changes in proteins
ionized by electrospray.

Figure No. 13 "Proteins sent by electrospray"

1991: Researchers Hennion, Ganem, Chait, and Katta discovered covalent complexes in
the metallic elements thanks to ionization by electrospray.

Figure No. 14 "Covalent Bonds"

1993: Chait, Wang, Beavis and Kent Piels, Zurcher, Sahcr, Moser, Fitzgerald, Zhu, and Smith achieve

create a sequence with MALDI for the exact identification of proteins as well as
oligonucleotides (DNA or RNA sequences with fifty base pairs or less)

Figure No. 15 "DNA Chain (oligonucleotides)"

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Heptacosa.

Infrared spectrometry is a type of absorption spectrometry that uses the infrared region.
of the electromagnetic spectrum. Like other spectroscopic techniques, it can be used to
identify a compound or investigate the composition of a sample.

This compound does not exhibit volatility at room temperature and presents a mass calibration range.
charge (m/z) of 614. Despite its molecular weight of 671 Daltons, the maximum ion it produces is
from 614 m/z in an electron impact ionization (EI) source. This may imply a limit for
the current software like Turbo Mass that has a limit of 1200 m/z.

Figure No. 16 "Fragmentogram of the Heptacosa"

Data of the Heptacosa:

Nombre comercial: Heptacosa-Acido- 10,12,17-triynedioico

Nombre: perfluorotri- butylamine (heptacosa, FC-43)
Molecular formula: C27H40O4
Peso Molecular: 428.6041 g/mol o 671 Da

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 Figure No. 17 "Molecular structure of Heptacosa"

PARTS OF THE MASS SPECTROMETER:

Figure No. 18 "Schematic of a mass spectrometer"

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 Figure No. 19 "Electronic Impact Source"

EXPERIMENTAL DEVELOPMENT:

Team Data:

Mass Spectrometer
Model Q-Mass 910
Perkin Elmer
Serial Number: E4AP19A00504
Date of Manufacture: May 1994
Part N°: N633-1300 120 V 50/60 Hz 12 A

Estatus del Equipo:

The mass spectrometer was found to be poorly assembled.

The mass spectrometer was found to be poorly coupled to the turbo pump.
Incorrect voltage and signal cables (opposite).

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Tools Used:
Pointed Pliers

Parakeet
Spanish key

Screwdriver and tacks

Figure No. 20 "Toolbox used for disassembly"

of the EM

PROCEDURE:

Disarmament of the Equipment:

Remove transfer line.

2. Remove the top cover of the mass spectrometer.
3. Open the front door of the spectrometer and remove the screws from the casing.

4. Place the metal casing on the table.

5. Remove the instrument used for the reference gas.
6. Disconnect voltage and signal cables from the EM.
7. Remove the neoprene supports located around the EM.
[Link] screws that hold the EM and take it out of the analysis area compartment.
9. Remove supports from the turbomolecular pump and the EM analyzer.
10. Disassemble the ion source and the analyzer.
11. Dismantle the ion detector.

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Equipment Assembly:

Assemble the ion detector.

2. Set up the ion source and the analyzer.
3. Place supports for the turbomolecular pump and the analyzer.
4. Place neoprene supports that are located around the EM.
5. Place the equipment in the analysis area and put in screws.
6. Connect the voltage and signal cables of the EM.

7. Place reference gas instrument.

8. Place the metal casing.
9. Close the front door and screw the metal casing.
10. Assemble the top cover of the mass spectrometer.
11. Reinsert the transfer line.

Q-MASS 910 PERKIN ELMER MASS SPECTROMETER:

Figure No. 21 "Exterior of the Spectrometer"

Dough

Figure No. 22 "Interior of the Spectrometer"

Dough

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Identification Table of Mass Spectrometer Parts

Mass Spectrometer
Parts of the Equipment Bibliography Definition Laboratory Definition

Input System. Input System.

In the input system, the sample is converted The laboratory has a team, a
to the gaseous state by heating to about gas spectrometer coupled to one of
400ºC and slowly introduced into the chamber of masses (CG/MS).
ionization. In the gas chromatograph is where separation occurs.

The purpose of the input system is to allow the the sample, to later proceed to
introduction of a representative sample in the mass spectrometer by means of a line
ion source with minimal vacuum loss. of transfer.

Figure No. 23 "Mass Spectrometer

Mass-Q910

Reference Gas. Reference Gas.

In order to have mass peaks In this equipment, the stored substance is the
known reference substances are used, heptacosa. This flask has a valve.
a rather complex spectrum but very good automatic that opens to regulate the flow of
known, to perform the calibration of the scale heptacosa towards the ionic source.
of the mass spectrometer. The substances of
Figure No. 24 'Glass Tube for Gas' The most commonly used reference is heptacosane for the The heptacosa is stored in a flask very
Reference calibration of weights in low equipment small glass that helps us do it

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 resolution and the PFK (perfluorokerosene) for those of tuning and calibration of the spectrometer
high. Normally what is done is to open the doughs.
vent valve to achieve a flow of
heptacosa towards the ionic source that causes the
permanent presence of the specter of this
substance with which the team compares the
mass spectrum detected with the one it has
stored in the computer's memory.
Ionization Source Ionization Source
Electrons are emitted by a filament. The ionization source is the medium through which the
tungsten or rhenium heated and are accelerated ions are charged with a certain level of energy and
for a potential of approximately 70 eV that in this way provoke fragmentation for
it is applied between the filament and the anode, the obtain complex or non-complex spectra.
trajectories of electrons and molecules The ion source of a mass spectrometer
they are at right angles and intersect in the center of convert the components of a sample into
the source, where they collide and take place the ions by electron bombardment, ions,
ionization. molecules or photons.
Figure No. 25 "Ionization Source"
The primary product is ions of a single Alternatively, ionization is carried out by
electronic impact
positive charge that is formed when the thermal or electrical energy.
high-energy electrons approach
sufficiently to the molecules so that
causing them to lose electrons due to repulsions
electrostatics

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 Mass Spectrometer Mass Spectrometer
The quadrupole instrument is a set of the The quadrupole is an ion analyzer that serves
four cylindrical bars that act as to separate them and transmit them to the detector, it has

electrodes. four bars, two connect to the positive pole and

The bars are electrically connected, a pair is connected to the negative pole, these bars are responsible for
connected to the positive pole of a variable source of carrying out the selection of the ions that can
direct current and the other pair connects to the pass to the detector. Generally, it only transmits
negative terminal. At each pair of bars ions from a small range of m/z relations.
potential alternating current variables of All the other ions are neutralized and are
radiofrequency, which are out of phase by 180°. eliminated. Generally, the instruments have
To obtain a mass spectrum with this a gold coating to avoid contamination of the
device, the ions are accelerated in space internal part.
between the bars with a potential of 5 to

Figure No. 26 "Mass Analyzer" 10 years.

Quadrupole Generally, quadrupole instruments

ions that differ in their mass are easily separated
in one unit.

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 Detector: Detector:
It is similar to the classical electron multiplier. The ions from the accelerator system
with the main difference that it does not have they arrive at the detector which is generally
multiple discrete dinodes, but it is consists of an emitter cathode that, upon receiving the
formed by a glass tube in the shape of impact produced by charged particles
horn, sometimes finished in a spiral shape or it emits electrons. These electrons are accelerated
snail, whose interior is coated by an oxide towards a dinode which emits several electrons
lead semiconductor composition and more upon receiving the impact of each electron. This
special characteristics. the process is repeated several times until obtained
The end of the detector tube is located at a a cascade of electrons reaching the collector
potential close to the ground, so there will be a achieving a strongly flowing current
continuous gradient of potential from the mouth to the amplified, by a procedure very similar to
detector background. Due to this, the electrons that is used in photomultiplier tubes. The
Torn away in the initial impact of the ion will fly in the obtained current that can be amplified again.
the direction of the less negative potentials, for electronic procedures and is taken to a
towards zero potential, resulting in a great register system.
number of impacts on the road, in each one

Figure No. 27 "Ion Detector" of which the number of will be multiplied

electrons. At the end of the tube's path
multiplier there is a collector cone with
the preamplification and amplification electronics
associated. Therefore, this detector is susceptible
as an employee both for detecting ions
positives as negatives.

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 Vacuum pump: Vacuum Pump:
Turbomolecular pumps consist of a The turbomolecular pump allows generating a
turbine with a series of blades that move at high vacuum in the system up to (10-8 torr), since for
a supersonic speed spinning at a carry out a good analysis in spectrometry
speed of 60000-90000 rpm. When the It is necessary to keep the team high for the masses.
gas molecules collide with the blades of the empty, this allows for formed lesions
rotor, receive an additional component of can move around without colliding with other ions and
pumping speed, in order to create a flow in this way register a good spectrum.
net macroscopic. The molecules driven by
the rotor must reach the stator blades
before colliding with another molecule, what the
it would deviate along its trajectory, so that it is fulfilled

Figure No. 28 "Turbomolecular pump" this condition, the speed of thermal agitation
of the molecules. The pressure must be
low enough for the journey
is greater than the distance between the rotor and stator. In

these conditions reach the flow condition

molecular and pumping starts to be efficient.

Table No. 1 "Parts of the Mass Spectrometer"

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 OBSERVATIONS:

Ramírez Anaya Adriel

The analyzer has neoprene supports on its exterior that serve as dissipators.
of effort when there is a high vacuum.

The container of laheptacosa is similar to a very small flask that helps us to

perform the calibration of the mass spectrometer since it is used as a reference gas.

The turbomolecular pump has blades that are used to generate high vacuum.
at least 10-8torr, which allows the molecules not to interact with the molecules
present in the air and we can obtain a better resolution of our spectrum.

The electronic impact where the sample enters has a gold coating to prevent it from
contaminate it when it dissipates into ions.

The quadrupole analyzer is a part composed of four rods arranged in a shape

two are connected to positive terminals and the other two to negative terminals with
the purpose of the ion flow to be selected and directed to the detector.

The ceramic detector has a trumpet shape where it receives ions upon collision.
deposited on a cathode.

CONCLUSIONS:

In this practice, we identify the parts that make up the mass spectrometer as well as its
functioning and the path the sample takes until it is analyzed, the identified equipment was a
gas chromatograph (GC) coupled to a mass spectrometer (MS) which through a
The transfer line carries the CG sample to the electronic impact of the EM.
In mass spectrometry, it is important to highlight that unlike chromatography, this provided
peak signals however in the masses the signals are fragments of the peaks.
On the other hand, the mass spectrometer consists of an input system, ionization source, the
mass analyzer, the detector and a turbomolecular pump, not to mention the gas compartment
for calibrating the equipment; all of them serve a specific function 'the separation
of molecular or atomic particles due to their different mass.

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Finally, we can say that the process of mass spectrometry basically includes
four stages:
Sample ionization.
Acceleration of ions by an electric field.
Dispersion of ions according to their mass/charge.
Detection of ions and production of the corresponding electrical signal.
And its application lies in that with this technique, it is possible to determine the molecular weight of all the

substances that can vaporize due to the position of the nozzle corresponding to the standard mass,
also to determine the molecular formula, if the instrument has high resolution it will be sufficient
precise determination of its molecular mass in order to attribute an empirical formula to it. Other times
it can be determined by the relationship between the heights of the peak corresponding to the standard mass and the

of the isotope peaks. On the other hand, also the identification of compounds by their
fragmentation pattern: the fragmentation of most molecules produces a large
number of peaks that allow the identification of numerous compounds and the recognition of
certain functional groups of them.

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OBSERVATIONS:

Sánchez Pino Javier Jefté

The mass system within the equipment only takes up 1/3 of the space since the rest is
composed of electronic circuits.
The electronic ionization team had a gold coating for its protection.
When conducting analysis, an air background must be done in order to rule out.
interferences in the desired fragmentogram.

CONCLUSIONS:

With the completion of the practice, the student checked the structure of a spectrometry team.
of masses coupled with a gas chromatograph as well as the space it specifically occupies
on your sample analyzer.
The equipment used in the practice was a Q-Mass 910 model mass spectrometer with a
quadruple analyzer, an electronic ionizer coated with gold to prevent corrosion, deterioration
in order to avoid contamination of the samples, a turbomolecular pump which can
reach a void of 10-5a 10-8torr (besides being a better option than differential pumps)
and a bulb with the reference gas.
By assembling and disassembling the equipment, the student confirmed the operation and importance of all the

parts of this qualitative and quantitative analysis instrument as well as why each piece has
a specific place when placed inside the analyzer, it was also verified why it exists
team union of instrumental analysis, since in this case the mass was coupled to a team of
gas chromatography because as such the mass cannot gasify the sample by itself
(for this model) which is a necessary parameter to achieve the fragmentogram.
Finally, returning to the part of the bulb with the reference gas, these devices use these
fluids in order to obtain known peaks in a fragmentogram and thus be able to calibrate them
correct way; for low-resolution mass spectrometers, heptacosa is used
what is a non-volatile fluid at room temperature with friendly characteristics for the equipment, is
to say, does not damage its internal parts, whereas for equipment that require a higher
precision and accuracy when analyzing samples such as high resolution is used
PFK (perfluorokerosene).

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OBSERVACIONES:

Anaya Flores Diana Laura

. The Perkin Elmer model Q Mass 910 mass spectrometer was found to be poorly assembled.
with the voltage and signal cables placed incorrectly.
. The team has neoprene, which dissipates vibrations and the impact of metals.
. The mass spectrometer 910 has a turbo molecular pump to create a vacuum.
coupled to another auxiliary pump.

. The team is connected to a gas chromatograph by a transfer line for the

separation, identification and quantification of mixtures of volatile and semi-volatile substances.

CONCLUSIONS:

Mass spectrometry is based on the generation of ions from molecules in the gaseous phase.
soda, to identify unknown compounds, to quantify known compounds and
elucidate the structure and chemical properties of the molecules.

Throughout this practice, the disassembly and reassembly of the mass spectrometer were carried out.

Q Mass 910 Perkin Elmer, for the identification of its parts as well as to carry out the
analysis of the path that molecules take by diffusion from the entry system to the
ion detector. The mass spectrometer mainly consists of the system of
input, the ion source, the mass analyzer, and the ion detector, the culés define the
type of spectrometer and the conditions of the system.

The analyte as a gaseous ion is studied in this way because of the movement of the ions in
The gaseous phase can be precisely controlled in electromagnetic fields and can be
easily manipulated, and also increases the sensitivity of the mass spectrometer.
The coupling of the gas chromatograph to the mass spectrometer allows for separation
compounds already in gas phase, as the ions enter in gas form in different fractions and
in different times continuously allows a sequential analysis of the samples.

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BIBLIOGRAPHY:

 Chapter 11. "Atomic Mass Spectrometry". Douglas A. Skoog, F. James Holler,

Timothy A. Nieman. Principles of Instrumental Analysis. 5th edition, Madrid Spain, Mc
Graw-Hill, 2001, p269 - 289
 Chapter 20. 'Molecular Mass Spectrometry'. Douglas A. Skoog, F. James Holler,
Timothy A. Nieman. Principles of Instrumental Analysis. 5th edition, Madrid Spain, Mc
Graw-Hill, 2001, p537 - 575
 Harvey, D., 2002, Modern Analytical Chemistry, Madrid: Mc Graw Hill /Interamericana
Spain s.a., 570 pgs.
 Siuzdak Gary, Mass Spectrometry for Biotechnology. Academic Press, Inc., USA, 1996

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