CLS 571 Fall 2020-21

Mass Spectrometry

American University of Science and Technology

Nina Esipenko, Ph.D.

1
 Mass Analyzers
Once the gas-phase ions have been produced, they need to be separated according
to their masses, which must be determined. The physical property of ions that is
measured by a mass analyser is their mass-to-charge ratio (m/z) rather than their
mass alone.

the separation of ions according to their mass-to-charge ratio can be based on

different principles:

2
 Mass Analyzers
main characteristics for measuring the performance of a mass analyser:

1. Mass range limit

The mass range determines the limit of m/z over which the mass analyser can
measure ions.
2. Analysis speed
also called the scan speed, is the rate at which the analyser measures over a
particular mass range
3. Transmission
ratio of the number of ions reaching the detector and the number of ions
entering the mass analyser. The transmission generally includes ion losses through
other sections of the analyser such as electric lenses before and after the analyser
4. Mass accuracy
It is the difference that is observed between the theoretical m/z (mtheoretical) and the
measured m/z (mmeasured). It can be expressed in millimass units (mmu) but is often
expressed in parts
5. Resolution
resolving power is the ability of a mass analyser to yield distinct signals for two ions
with a small m/z difference
3
 Magnetic deflection (sector)

EI ionization

4
 Magnetic deflection (sector)
 Separation in this way is effected by the application of a magnetic field
perpendicular to the motion of the ions leaving the ion-source.

 The trajectory of the ion depends on the applied forces: The Lorentz force
(FL) and the centrifugal force (FC).
B - strength of the magnetic field,
z - amount of charges,
e - charge of one electron
v – velocity
r – curvature of a radial path
m – ion mass

For every value of B, the ions with the same

charge and the same momentum (mv) have a
circular trajectory with a characteristic r value.

Thus, the magnetic analyser selects the ions

according to their momentum.

mv = Bzer 5
 Time-of-Flight (TOF)
 Ions are produced within the ion source
 Ions are subjected to an accelerating voltage
 Ions “drift” or “fly” down an evacuated tube of a set length L
 Ones free from the region of accelerating voltage the speed at which the ions
travel down the tube is dependent upon their mass and charge (heavier ions
have larger arrival times compared to lighter ions )
 Scanning the mass range of all ions is very rapid

6
 Time Of Flight (TOF)
ions are separated on basis of the time t needed to travel a path L:

t – time of flight (drift time)

m – ion mass
z – ion charge
eV – electric potential difference (voltage)
L – length of the path (drift length)

Advantages:
 All ions detected at once
 High mass accuracy
 Reasonable performance for the cost
 Theoretically unlimited mass range
 Well suited for pulsed ionization
methods (MALDI-TOF)
Disadvantages:
 High vacuum required for resolution and
accuracy (complex vacuum system
necessary)
 Must be often recalibrated (temperature
and voltage fluctuations alter flight times)
 Long flight tubes for high resolving power
can make instruments large 7
 TOF
Linear TOF (LTOF) Reflectron TOF (RTOF)
(discussed in previous slides)

The problem with linear detector: Peaks are inherently broad; poor mass resolution

The cause: Ions of the same mass coming from the target
have different speeds. This is due to uneven energy
distribution when the ions are formed by the laser pulse.

8
 Reflectron Time Of Flight (RTOF)
 TOF is equipped with a reflectron, or an ion mirror
 The reflectron deflects the ion using an electric field and increases the path length,
improving signal resolution
 The most rapid ions (i.e. higher energies), will penetrate deeper into the decelerating
zone than low energy ions
 The ions of the same mass are time focused, then will arrive on the microchannel
detector at the same time (leading to a higher resolution)

L1
(ion mirror)

+ +
+ + ++
+
Acceleration region

+++
++++
++++
L2
9
 Quadrupole
 Quadrupole made up of four parallel rods
 The four road are electrodes, with electric field around them
 Electrodes have voltages applied to them (RF and DC voltages)
 Each pair of rods are connected and have the same voltage
 Depending on the voltage magnitude only ion of a specific m/z make it through
quadrupole

+
- -
+

10
Ions travelling along the z axis are subjected to the influence of a total electric field
Ion separation is performed by using controlled voltages applied to the mass analyser
rods which impact electrostatic field inside the devise
 Only ions of a certain mass-to-charge
ratio will reach the detector for a given
ratio of voltages (stable trajectory)

 other ions have unstable trajectories

and will collide with the rods.

 This permits selection of an ion with a

particular m/z

 allows the operator to scan for a

range of m/z-values by continuously
varying the applied voltage

11
 Tandem MS/MS
tandem mass spectrometer can be
conceived in two ways:

performing tandem mass spectrometry performing an appropriate

in space by the coupling of two sequence of events in an ion
physically distinct instruments storage device

Quadrupole – TOF quadrupoles as analysers

Magnetic sector – quadrupole Known as triple quadrupole,
etc. QqQ, or Q1-Q2-Q3

12
 Tandem MS/MS
MS/MS is the combination of two or more MS experiments.
 To get structural information by fragmenting the ions isolated during the first
experiment
 To achieve better selectivity and sensitivity for quantitative analysis by selecting
representative ion transition

Ionization

13
 Tandem MS/MS
• A single m/z is selected by the quadrupole analyzer (Q1).
• Fragmentation occurs in the collision cell (q2).
• The 2nd quadrupole analyzer (Q3) can scan or select a single m/z.

Q1

Q2

Q3
 Tandem MS/MS

Single MS Scan Single Ion Monitoring (SIM)

15
Tandem MS/MS

16
 Tandem MS/MS

Collision Cell (q2)

1st set of resolving 2nd set of resolving

quadrupole rods (Q1) quadrupole rods (Q3)

17
 R

127

229
18