Bonga university

Instrumental Analysis II
(Chem.2052)

Chapter 8
Mass spectroscopy (MS)
2012 E.C
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 Chapter seven
Mass Spectrometry
A technique for measuring and analyzing molecules, that involves
introducing enough energy into (neutral) target molecule to cause its
a
ionization and disintegration. The resulting primary ions and their
fragments are then analyzed, based on their mass/ charge ratios, to produce
a "molecular fingerprint."

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3 3
 MS does involve a specific region of the
electromagnetic
not spectrum (because it is not directly
interested in the energies of emitted photons, electronic or
vibrational transitions, nuclear spin transitions, etc…)

The mass spectrometer is an instrument that separates

gas phase ionized atoms, molecules, and fragments of
molecules by the difference in their mass-to-charge ratios.
i.e. Mass Spectrometry– a method of separating and
analyzing ions by their mass-to-charge ratio
The mass-to-charge ratio is symbolized by m/z, where the
mass m is expressed in atomic mass units and z is the number of
charges on the ion.

 Mass-to-charge ratio (m/z) is the ratio of the mass of an ion

(m) to its charge (z). The term z symbolizes the number of
charges on the ion; this number may be positive or negative, such
as +1, -1, +2, +10, and so on. The number of charges is not the
same as the total charge of the ion in coulombs.

The total charge q = ze, where e is the magnitude of the charge

on the electron, 1.6 x 10-19C
While some MS methods do generate mostly +1 charged
ions, many new techniques generate ions with multiple
charges.
 Advantages of MS
Is used to measure the relative molecular weight and
consequently the molecular formula.
It is used to detect the fragments – C-C-C-C-O-C- by
combining the fragments, we can arrive at the possible
molecular formula.
Small amount of sample can be analyzed of low
detection limit.
Is the only tool to determine the molecular weight of the
molecule.
 Disadvantages of MS

Sample cannot be
recovered.
High cost of the instrument.
Mass spectrum
 A plot of abundance vs m/z is called a 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.
Amass spectrum of cocaine
 The most abundant ion (tallest peak is called base peak.
The ion given as a result of removal of a single electron
from a molecule is known as Molecular ions, designated by
M+.. It is positively charged b/c it lost electrons. It is an ion
consisting of essentially the whole molecule
 Ignoring the heavy isotope, the m/e ratio of M+. It is
found to be at the far right of the chart.
Example: C2H5OH neglect 17O or 18O
and 13C or 14C

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A block diagram of a “generic” mass
spectrometer:
Ionization Sources
 E l e c t r o n I o n i z a t i o n (E I )
Gas Phase
 Chemical Ionization (CI/APCI)
 Photo-ionization (APPI)
 Electrospray (ESI)
 Matrix-assisted Laser Desorption ( M A L D I )
 Field Desorption (FD) D esorption
 Plasma Desorption (PD)
 F a s t a t o m b o m b a r d m e n t ( FA B )
 High-temperature Plasma (ICP)

Io n i z a tio n Mass
Detector
Source A n a ly z e r

S e e a l s o Ta b le 2 0 - 1 in S k o o g , e t al.

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Electronionization (EI):
One method of forming ions from sample molecules or
atoms is to bombard the sample with electrons is called
electron ionization (EI):

Where: M is the analyte molecule; e- the electron;

and M+. the ionized analyte molecule; this species is called
the molecular ion.
 Molecular ion M+.
 Is a radical cation formed by the loss of one electron.
Has the same mass as the neutral molecule, because
the loss in mass of one electron is too small to measure.
 In most cases, molecular ions have sufficient energy
as a result of the ionization process to undergo
fragmentation to form other ions of lower
m/z.
 Unfortunately, in EI spectra, the molecular ion is often
of low abundance and is not always observed
 Used to calculate number of C, O, N….present
 The molecular ion is always an odd electron ion.
Hence, the molecular ion is never an even electron
ion.
N.B-Aromatic compounds and conjugated
hydrocarbons give more intense molecular ion
peaks than alkanes; aliphatic alcohols, nitrates,
and highly branched compounds tend not to
give peaks for the molecular ion
Nitrogen Rule
The m/z value of the molecular ion is always an even
number if the molecular ion contains either no nitrogen
atoms or an even number of nitrogen atoms.
 If the molecular ion contains an odd number of
nitrogen atoms, the m/z value (and the molecular
weight) must be an odd number.
This is a very important rule to remember and it is valid
for organic compounds containing C, H, N, O, S, the
halogens, P, Si and many other elements. If the highest m/z
value in a mass spectrum is an odd number, the ion must
contain an odd number of nitrogen atoms if it is the
molecular ion. 17 17
18 18
 Components Of A Mass Spectrometer

Ionisation Ion Separation Ion Detection

Ion Mass Analyser Detector
Source Quadrupole Electron Multiplier
Electron Magnetic Sector Field Multichannel plate
Ionisation (EI)
Electric Sector Field Faraday Cup
Chemical
Ionisation (CI) Time-Of-Flight (TOF)

Fast Atom Bombardment (FAB) Ion Trap

Electrospray Ionisation (ESI)

 Why they are kept in high vacuum? It is to prevent
the formation of secondary ions M++
 All mass spectrometers require
 a sample input system,
 an ionization source,
 a mass analyzer,
 and a detector.
Fragmentation
 The energy of the electron responsible for the ionisation
process must be sufficient to knock out an electron
 Typically about 10-12 eV, is known as the appearance
potential.
 In practice much higher energies (-70 eV) are used
 Large excess energy (1 eV = 95 kJ moll) causes further
fragmentation of the molecular ion-break apart.
 The fragments formed may be
 ions,
 neutral molecules,
 radicals, and the like.
Fragments may undergo more fragmentation into even
smaller pieces.
The mass and abundance of these fragments are
called the molecule’s fragmentation pattern.
Each molecule has its own characteristic
fragmentation pattern under particular ionization
conditions.
The possible mass loses are:
Mass loss of 4 – 14 and 21- 25 are uncommon, there may be
1, 2, 3 hydrogen loss but there is no 4 H loss. There could
be CH3 OH H2O
15 17 18
C2H4 CO CHO
28 28 29
 There could be (M-15) +. = M- CH3 , (M-35) +. = M- Cl
, (M-17) +. = M- OH, (M-18) +. = M- H2O, (M-28) +. =
M- CO or C2H4
 Fragmentation Pattern

 As only species bearing a positive charge will be detected,

the mass spectrum will show signals due not only to [M]+.
but also due to A+, C+. and to fragment ions resulting from
subsequent fragmentation of A+ and C+·
 As any species may fragment in a variety of ways, the
typical mass spectrum consists of many signals. The mass
spectrum consists of a plot of masses of ions against
their relative abundance.
Table8.1 Common Neutral Losses from the
Molecular ion
Fragmentation Processes
Three factors dominate the fragmentation processes:
a. Weak bonds tend to be broken most easily
b. Stable fragments (not only ions, but also the
accompanying radicals and molecules) tend to be
formed most readily
c.Some fragmentation processes depend on the ability of
molecules to assume cyclic transition states.
Favourable fragmentation processes naturally occur
more often and ions thus formed give rise to strong
peaks in the mass spectrum
1. Cleavage at Branch Points.
 Cleavage of aliphatic carbon skeletons at branch points is
favoured as it leads to more
2. β- Cleavage.
(Chain cleavage tends to occur β to heteroatoms, double
bonds and aromatic rings because relatively stable,
delocalised carbocations result in each case.
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3. Cleavage α to carbonyl groups.
 Cleavage tends to occur α to carbonyl groups to give stable
acylium cations. R may be an alkyl, -OH or -OR
group.

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4. Cleavage α to heteroatoms.
 Cleavage of chains may also occur α to heteroatoms, e.g. In
the case of ethers

5. Retro Diels-Alder reaction Cyclohexene derivatives

may undergo a retro Diels-Alder rxn
6. The McLafferty rearrangement

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 Summary of fragmentation pattern by
functional groups
1. Alkane
 Peak will appear commonly at m/e = 15, 29,
43, 57,............ Peak at 15 is very common for
saturated hydrocarbons.
i.e. m/e = 15 is for CH3, m/e = 29 is for CH2CH3,
m/e = 43 is for CH2CH2CH3, m/e = 57 is for
CH2CH2CH2CH3

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 Examples:
 Mass spectrum of n-butane
35 35
Mass spectrum of 2,2,4-trimethylpentane
2. Alcohol
 m/e = M-1, M-2, M-3, M-17 and M-18 are
common especially M-1 and M-18 must exist
3.
Alkenes
cleavage to the double is the most common
Example
 (CH2=CHCH2CH2CH3)+. Due to the
resonance,
+ .
cleavage will give us CH2=CHCH2
and
CH2CH3 ( at
+
m/e = M-29, CH2=CHCH2 stable
ion).
4. Ethers
– are dominated by cleavage to the hetroatom (oxygen)

Path 2, Path1
Example: CH3-CH2 - CH-O-CH2 - CH3
path3
CH3

There are three possibilities, which one is the most

dominant? It is path2 b/c the unpaired electron is
found at CH2 but at path 1 and path3 the unpaired electron
is found at CH3 group. i.e. Besides the O effect,. the
Loss of longest chain is more common: CH3-CH2
branch
+CH-O- CH - CH
+
have effect2 ⇒ C2
3
then C3 and then C1.
CH3 43 43
44 44
6. Ketone
Example: Mass spectrum of 2-butanone
 7.
–Aromatics
are characterized by few peaks m/e = 91, 65, &
39 and 92, 77 & 51
Molecular Formulae and Isotopic Abundances
 The two stable isotopes of carbon are 12C and 13C.
 The12C constitutes 98.90% of all naturally occurring
carbon atoms; 13C is 1.10% of all naturally occurring
carbon.
 Therefore, in a compound containing one carbon atom,
98.90% of the molecules will have a 12C atom while
1.10% will have a 13C atom.
 In every organic compound (natural or synthesized
from natural sources), the 13C will result in a peak that is
one mass number greater than the mass of the molecular
ion.
 This 13C-containing peak is generally designated as M+
1.
If our molecule contains only carbon and hydrogen, since
there is 1.1% relative natural abundance of 13C compared to
100% 12C, and our mass spectrum shows a 1.1%
abundance of M+1 to M, only one carbon atom can be
present in the molecule.

That is, if we are dealing with a hydrocarbon and only

one carbon atom is present, then the ratio (M+ 1)/M =
1.1%.

If two carbons are present in the molecule, as there are in

ethane, the probability of 13C being present is twice as great
and (M+ 1)/M = 2.2%.
Counting Carbon Atoms
For compounds containing carbon and hydrogen
There is a definite relationship between the
peaks at M + 1 and M that is directly related to
the number of carbons present in hydrocarbon
molecules:
Example: Spectrum of benzene
• For benzene, C6H6, the molecular ion m/z =78, and the (M+1) m/z =
79. The relative abundance of the (M+ 1) peak is 6.4%; dividing
6.4% by 1.1% confirms that there are 6 carbons in the
molecule.
Counting Carbon, Nitrogen, and Sulfur Atoms
 Not only carbon, but also other elements contribute to the
M+1 peak intensity, notably nitrogen and sulphur when they
are present.
Silicon obviously will impact the M+1 peak in
organosilicon compounds such as silicone polymers.
 A more general equation for the ratio of the M+1 to M peak
is:

 Where #C atoms is the number of carbon atoms in the

molecule, and so on
Counting Oxygen Atoms
Similarly, the M+2 peak contains intensity
contributions from oxygen, Cland Br,and
heavy isotopes of carbon, hydrogen, and so on. multiple
 A general formula for compounds that contain only
C, H, N, O, F, P, and I can be used to ascertain the
number of oxygen atoms, in theory.
Oxygen has two important isotopes, 16O and 18O, with
a relative abundance 18 O/16O of 0.2%.
 In reality the 18O isotope is of low abundance and
oxygen- containing compounds often fragment so that the
molecular ion is of low intensity or not detected.
 In practice, the observation of isotope information for
oxygen is often difficult.
The number of oxygen atoms in a molecular ion can be
calculated from, ignoring hydrogen:
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