TOF MS

• In a crude sense, TOF is similar to chromatography,…

• … except there is no stationary/ mobile phase, instead

the separation is based on the kinetic energy &
velocity of the ions.
TOF system. As time evolves, the ions
(formed at the source) are separated.
Instrument Design: The Time-of-Flight Mass
Spectrometer
 Instrument Design: The Time-of-Flight Mass
Spectrometer

• If all ions obtain the same K.E, the ions of lesser m/z will
have greater velocity than ions of greater m/z.

• Therefore, as ions traverse the analyzer, they separate

in space.

• A detector is positioned at the end of the analyzer to

measure the arrival time of ions.
• Ions of lesser m/z arrive first, followed by ions of
greater m/z.
Instrument Design: The Time-of-Flight Mass
Spectrometer

• A plot of intensity or abundance versus time is

made to show the arrival time distribution of the
ions detected (see Figure nxt slide).

• The plot of abundance versus time is calibrated

& replotted as intensity versus m/z
Instrument Design: The Time-of-Flight Mass
Spectrometer
Instrument Design: The Time-of-Flight Mass Spectrometer

• The lighter fragments travel through the tube

faster than the heavier fragments,..

• .. & the different masses are detected at different

times, depending on their time of flight through
the tube.
 The Time-of-Flight Mass Spectrometer
• Ions of the same charges have equal kinetic energies;
• kinetic energy of the ion in the flight tube is equal to the
kinetic energy of the ion as it leaves the ion source:

KE=mv2=zV

• The time of flight, or time it takes for the ion to travel the
length of the flight tube is:
Tf=L/v
with L is the length of tube & v is the velocity of the ion
 Instrument Design: The Time-of-Flight Mass Spectrometer

The signal is thus measured vs. time, but can be translated into the traditional
mass spectrum.
 Different Types of MS
• ESI-QTOF
– Electrospray ionization source + quadrupole
mass filter + time-of-flight mass analyzer
• MALDI-QTOF
– Matrix-assisted laser desorption ionization +
quadrupole + time-of-flight mass analyzer

10
 Different Types of MS
• GC-MS - Gas Chromatography MS
– separates volatile compounds in gas column
and ID’s by mass
• LC-MS - Liquid Chromatography MS
– separates delicate compounds in HPLC
column and ID’s by mass
• MS-MS - Tandem Mass Spectrometry
– separates compound fragments by magnetic
field and ID’s by mass

11
 What is MSMS?
MS/MS means using two mass analyzers (combined in
one instrument) to select an analyte (ion) from a
mixture, then generate fragments from it to give
structural information.
mass analyzers are used in tandem.

Mixture of Single Fragments

ions ion

Ion
MS-1 MS-2
source
 Tandem Mass Spectrometry
• Purpose is to fragment ions from parent ion to provide
structural information about a molecule
• Also allows separation and identification of compounds
in complex mixtures

• Uses two or more mass analyzers/filters separated by a

collision cell filled with Argon or Xenon

• Collision cell is where selected ions are sent for further

fragmentation

13
 Tandem Mass Spectrometry
 one mass analyzer is used to “select” a
particular analyte ion from those that enter it.

 the magnetic field strength in the first mass

analyzer is set (not scanned) such that only the
selected ion is allowed through.

 This ion then undergoes fragmentation in a

“collision cell.”
 Tandem Mass Spectrometry
 ionized gas (Ar, He, or N2) is pumped into this collision
cell at low pressure.
 Analyte ions colliding with the charged gas particles
(hence the name “collision cell”) are energetic enough to
generate fragments.
 The “product ions” so generated enter the second mass
analyzer such that a complete mass spectrum of the
analyte is generated in the usual way.
 LC-MS/MS and GC-MS/MS are common designations
for the complete procedure.
 Tandem Mass Spectrometry

 Sample molecules emerging from the LC column are ionized via an

electrospray ion source.
 The first mass analyzer selects the analyte ion to be passed on to the
collision cell.
 The ions from the collision cell then pass through the second mass
analyzer/ion detector where the normal mass spectrum is measured.
 Tandem Mass Spectrometry
• Different MS-MS configurations
– Quadrupole-quadrupole (low energy)
– Magnetic sector-quadrupole (high)
– Quadrupole-time-of-flight (low energy)
– Time-of-flight-time-of-flight (low energy)

• Fragmentation experiments may also be performed on

single analyzer instruments such as ion trap instruments
& TOF instruments

17
 Different MS-MS Modes
• Product or Daughter Ion Scanning
– first analyzer selects ion for further fragmentation
– most often used for peptide sequencing
• Precursor or Parent Ion Scanning
– no first filtering, used for glycosylation studies
• Neutral Loss Scanning
– selects for ions of one chemical type (COOH, OH)
• Selected/Multiple Reaction Monitoring
– selects for known, well characterized ions only

18
 Summary: acquiring a mass spectrum

Ionization Mass Sorting (filtering) Detection

Ion Ion
Source Mass Analyzer Detector
Form ions
Sort Ions by Mass (m/z) Detect ions
(charged molecules)

100

75

Inlet • Solid 50

• Liquid 25

• Vapor 0
1330 1340 1350

Mass Spectrum
 Mass Spectroscopy: mass spectrum

 A plot of abundance versus m/z is called a mass

spectrum.

 The mass spectral data can be shown as a plot

or as a table.

 At specific m/z ratios, sharp peaks are

displayed, each representing a particular
fragment having that ratio

20
Mass Spectroscopy: mass spectrum

 The key is that a given molecule always disintegrates

into the same fragments..

 … & therefore always displays the same mass

spectrum, a molecular fingerprint.
 This occurs because certain bonds within a molecule
are weaker than others.
 The mass spectral pattern is due to the fact that it is
these same bonds that always break—and thus the
same fragments always form—when subjected to
the bombardment of the electron beam.

21
Mass Spectroscopy: mass spectrum

 Also noteworthy is the fact that different naturally

occurring isotopes of a given element are always
present in the various fragments & are thus always
apparent in the mass spectrum.!

 A digitized mass spectrum of benzene, C6H6, is

shown in the next slide

22
Mass Spectroscopy: mass spectrum

23
 Mass Spectroscopy: mass spectrum

 The most abundant peak in the spectrum, called the base peak,
is scaled to 100,..
 … so the y-axis represents the relative abundance of the ions of
each m/z value.

 The nominal mass of the benzene molecular ion is equal to the

sum of the nominal masses of the C & H isotopes, ..

 …so 6(12) + 6(1) = 78; the most abundant peak in this benzene
mass spectrum is the molecular ion peak at m/z 78.

 A # of fragment ions at lower values of m/z are also seen in this

spectrum.

24
 Mass Spectroscopy: mass spectrum

mass spectral data for benzene as shown in the plot.

25
 A mass spectrum of cocaine
 This example shows that the molecular
ion is not always the most abundant ion!
The m/z values &
the fragmentation
pattern enable the
analyst to
determine the MW
and structure of
organic
compounds by MS.

26
 Mass Spectroscopy: mass spectrum

 Another simple example is the mass spectrum of

methane, CH4
 CH4 molecule breaks into the following
fragments (in order of relative abundance):,
CH4+, CH3+ CH2+ & 13CH4+.

 In the figure, each peak is identified as being

due to one of these fragments.

27
The mass spectrum of methane.

28
Mass Spectroscopy: mass spectrum

 Notice that one of the peaks is due to a fragment with

the 13C isotope (the fragment identified as 13CH4+).

 A 13C atom is one of about every one hundred

carbon atoms in nature.

29
 Mass Spectroscopy: mass spectrum

 Because the fragmentation pattern produced by a mass

spectrometer can be used as a fingerprint of molecule,…

 …. the mass spectrum reveals, for example, whether the

correct compound has been synthesized and whether
contaminants are present.

 One can see that it is a molecular fingerprint, just as

absorption spectra are molecular fingerprints, and that it
is a powerful tool for identification purposes.

30
 O
CH3
MW 194

H3C
N
N

N
O N

CH3
 O
CH3

H3C
N
N

H 109 m/z
N
O N

CH3
 O
CH3

H3C
N
N

N
O N

55 m/z
CH3
 Mass Spectroscopy

 Besides the obviously significant applicability to

molecular compound identification,..

 .. MS also finds application in elemental analysis, such

as to determine:
 what isotopes of an element might be present in a
sample or,..
 when combined with inductively coupled plasma
(ICP) what the conc is of an element in a sample.

35
Mass Spectroscopy

 The isotopes of radioactive elements may be

detected in a variety of samples, including biological
matrices.

 Different isotopes are detected because the differing

counts of neutrons that characterize atoms of
different isotopes produce ions of different m/z in the
mass spectrometer.

36
Masses in MS
 Monoisotopic mass is
the mass determined
using the masses of
the most abundant
isotopes
 Average mass is the
abundance weighted
mass of all isotopic
components

37
 The mass spectrum of elemental lead.

Isotopes of Pb with atomic weights of 204, 206, 207, and 208 are
known and have the relative abundance indicated by the signal
heights 38
 Ionization Sources for Inorganic MS

 The following ionization sources are used mainly in

inorganic (atomic) MS, where the elemental
composition of the sample is desired.

 The glow discharge (GD) & spark sources are used

for solid samples, while the inductively coupled
plasma (ICP) is used for solutions.

 All three sources are also used as atomic emission

spectroscopy sources..where they ve been
discussed in detail

39
Inductively coupled plasma
 The most commonly used techniques for the
determination of trace concentrations of elements
in samples are based on atomic emission
spectrometry (AES).

 To dissociate sample molecules into free atoms,

thermal sources such as flames, furnaces &
electrical discharges are used.

40
Inductively coupled plasma
 More recently, other types of electrical discharges, namely
plasmas have been used as atomization/ excitation sources
for AES.
 These techniques include inductively coupled plasma (ICP) &
direct coupled plasma (DCP).

 Plasma: a hot ionized gas made up of ions & electrons that is

found in the Sun, stars, and fusion reactors.

 Plasma is a good conductor of electricity and reacts to a magnetic

field, but otherwise has properties similar to those of a gas.

41
ICP
 The primary goal of ICP is to get elements to emit
characteristic wavelength specific light which can
then measured.

 An ICP is a very high temp (7000-8000k) excitation

source that efficiently desolvates, vaporizes,
excites, and ionises atoms.

 Molecular interferences are greatly reduced with

this excitation source but are not eliminated
completely.

42
 ICP
 An ICP requires that the elements which are to
be analysed be in solution.

 An aqueous solution is preferred over an organic

solution, as organic solutions require special
manipulation prior to injection into the ICP.

43
 MS Combined with Inductively Coupled Plasma

 The ICP technique described under emission spec can

be coupled with MS.

 This source is used primarily for liquids.

 Solid samples are generally dissolved in acid and diluted

for analysis; organic solvents can also be used.

 The source produces ions from the elements introduced

into the plasma as well as radiation; these ions can be
extracted into a mass analyzer.

44
What is ICP-MS?

 Employs plasma as ionization source & a mass

spectrometer (MS) analyser to detect ions

 It can perform qualitative & quantitative

analysis.

 ICP-MS commercially introduced in 1983 and has

gained general acceptance in many types of
laboratories.

45
Reasons for the growing popularity

•Performs multi-elemental analysis with

excellent sensitivity and high sample
throughput
• Instrument detection limits are at or below
ppt level
• Productivity is unsurpassed by any other
technique
• Isotopic analysis (one isotope or ratio)
Elements measured by ICP-MS
 Elements measured by ICP-MS
• The ICP-MS instrument measures most of the
elements in the periodic table.

• The elements shown in color in prev fig can be

analyzed by ICP-MS with detection limits at or below
the ppt range.

• Elements that are in white are either not measurable

by ICP-MS or do not have naturally occurring
isotopes.
 ICP-MS
• For reasons that often involve human health,
knowing the isotopic composition of a sample
can be highly important.

• only ICPMS is used routinely for determining

isotopic composition. !!
 How does ICP-MS work?
•Samples introduced into an
argon plasma as aerosol
droplets.
• The plasma dries the aerosol,
dissociates the molecules, and
then removes an electron from
the components, forming singly-
charged ions, Figure 1. The ICP Torch showing the
fate of the sample (PerkinElmer,
•Ions are directed into a mass Inc.)

filtering device known as the

mass spectrometer.
 An ICP-MS consists of
• Sample introduction system
• ICP torch and RF coil – generates the argon plasma,
which serves as the ion source of the ICP-MS
• Interface – links the atmospheric pressure ICP ion
source to the high vacuum mass spectrometer
• Vacuum system – provides high vacuum for ion
optics, quadrupole, and detector
• Collision/reaction cell – precedes the mass
spectrometer and is used to remove interferences that
can degrade the detection limits achieved.
 An ICP-MS consists of
• Ion optics – guides the desired ions into the
quadrupole while assuring that neutral species
and photons are discarded from the ion beam.
• Mass spectrometer – acts as a mass filter to sort
ions by their mass-to-charge ratio (m/z)
• Detector – counts individual ions exiting the
quadrupole
• Data handling and system controller – controls all
aspects of instrument control and data handling
to obtain final concentration results.
 ICP-MS instrumentation and
principle
Detector
Plasma generates (e.g.
Under vacuum
electron
positive ions
multiplier)

nebuliser
Interface
Sorted by mass analyser,
Spray chamber e.g. quadrupole,
magnetic sector,
according to m/z ratio

sample

http://www.cee.vt.edu/ewr/environmental/teach/smprimer/icpms/icpms.htm
MS Combined with Inductively Coupled Plasma

 The ICP torch is usually mounted horizontally with the tip of the
plasma at the entrance to the mass analyzer

The “exhaust” of the ICP source is fed into the mass

spectrometer for analysis by the mass analyzer portion
of the mass spectrometer

54
55
 ICP-MS
 Extracting ions from the plasma can
present a major technical problem in
ICPMS.
 While an ICP operates at atmospheric
pressure, a MS operates at high
vacuum, typically less than 10-6 torr.

 The interface region between the ICP

& the MS is thus critical to ensure
that a substantial fraction of the ions
produced are transported into the
56
ICP-MS
 Theinterface usually consists of two
metal cones, called the sampler & 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 MS has

about the same ionic composition as the
plasma region from which the ions are
extracted
57
ICP-MS
 ions from the plasma enter into the
mass spectrometer through a two-
stage interface.

 The plasma is centered on the sampler

cone, and ions and plasma gas pass
through the orifice in the cone into a
vacuum-pumped region.

 Most of the Ar gas is pumped away in

this region.
58
ICP-MS
 The remaining ions pass through
the skimmer cone into the mass
spectrometer.
 The skimmer cone is a sharper-
angled cone with an orifice of about
0.9 mm in diameter.

59
ICP-MS
 The ICP has a high ionization
efficiency, which approaches 100%
for most of the elements, & it
produces mainly singly charged
+ve ions.

 The mass spectra therefore are

very simple, & elements are easily
identified from the masses and the
isotope ratios easily measure

60
ICP-MS Spectra
ICPMS spectrum consists of a simple series of isotope peaks for
each element present along with some background ionic peaks.


61
ICP-MS
 The ability to rapidly analyze solutions
of dissolved inorganic materials (metals,
rocks, bones, ceramics, ash, and the
like),…
 … combined with the simple mass
spectra and low background and
excellent sensitivity, led to the rapid
spread in the use of ICP-MS in many
fields.

62
 ICP-MS
schematic diagram of an ICP-MS system with a
quadrupole mass analyzer.

63
 ICP-MS

Layout of a new ICP-MS, the PerkinElmer NexION 300, with the plasma vertical
to the MS
64
ICP-MS
 instrument has a quadrupole ion deflector
(QID) above the torch.

 The QID turns positively charged ions from

the plasma 90° to the left, into the mass
analyzer
 nonionized material flows straight up and out
of the system

65
 Sample introduction
system
• provides the means of getting samples into the
instrument
• composed of a nebulizer and spray chamber

• The liquid sample may be introduced to a

nebulizer by a peristaltic pump or through self
aspiration that creates an aerosol of fine droplets.
• The fine droplets are passed through a spray
chamber before they are allowed to enter the
plasma
 The vacuum system
• provides correct operating pressure
• The distance from the interface to the
detector <=1 meter
• ions need not to collide with gas molecules
during the travel
• so gas molecules are removed by using a
combination of a turbo molecular pump and
mechanical roughing pump
 The detector – counting ions
• The ions exiting the mass spectrometer strike
the active surface of the detector (dynode) and
generate a measurable electronic signal.
• a dynode, releases an electron each time an ion
strikes it.
• released electrons strike a second dynode where
more electrons are released until a measurable
pulse is created. By counting the pulses
generated by the detector, the system counts the
original ions.
 Data handling and system controller
• The software compares the intensities of the
measured pulses to those from standards, which
make up the calibration curve, to determine the
concentration of the element.
• The software translates the ion counts measured
by the detector into information
• provide data in one of four ways – semi-
quantitative analysis, quantitative analysis,
isotope dilution analysis, and isotope ratio
analysis.
liquid samples limitations
• ICP-MS has some limitations as to the amount of
total dissolved solids in the samples.
• Generally, no more than 0.2% TDS for best
instrument performance and stability
• many sample types, including digested soil and
rock samples must be diluted
• If samples with very high TDS levels are run, the
orifices in the cones will eventually become
blocked, causing decreased sensitivity and
detection capability and requiring the system to be
shut down for maintenance.
 Approximate costs

Costs depends mainly on

Number of samples
Number of elements to be
analyzed
Time required
MS Combined with Inductively Coupled Plasma

 The intensity of the signal due to the analyte ion is

proportional to the concentration of the analyte in the
sample.

 In most cases there will be more than one analyte ion

for a given element (due to the presence of more than
one naturally occurring isotope) and therefore more
than one signal for that element.

73
 Applications
Elements and isotopes in
• Soil
• Water/wastewater
• Food
• Human and biological samples
• Industrial like pharmaceutical………..
Comparison: AAS, ICP-OES, and ICP-
MS
 AAS: Single element, ppm/ppb range
 Cheap, simple
 Small dynamic range
 GFAAS about 100 times more sensitive than FAAS,
but also more challenging
 ICP-OES: Multi-element, ppb range
 Limited spectral interferences, good stability, low
matrix effects
 ICP-MS: Multi-element, possible to reach ppt
(or even ppq, Part per quadrillion )
 Most complex, most expensive, lowest detection
limits, isotope analysis possible