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M.

Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

Mass spectroscopy
Mass spectroscopy is a quantitative and qualitative analytical technique by
which we can measure the molecular mass and formula of a compound and
the record is known as mass spectra.

Mass spectra is useful −


 To establish the structure of a new compound
 To give the exact molecular mass
 To give the molecular formula
 To indicate the presence of functional group in a compound

Principle/function:
The mass spectrometer is designed to perform four basic functions −
• To vaporize the compound by increasing volatility.
• To generate the ions from the neutral compound in resulting vapor
pressure
• To separate the ions according to their mass to charge ratio (m/z) in
a magnetic field.
• To collect the mass and record.

Theory:

1. Molecular ion production:


 Mass spectrometer is a device for the production and weighing
of ions.
 Molecules are subjected to bombardment by a stream of high-
energy electrons, converting some of the molecules to ions. The
molecular ions are usually radical cation and some may be
radical anion.

- e- + - e- -
[M] [M] or, [M] [M]

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

2. Fragmentation:
 When the molecule has been bombarded by high-energy
electrons to produce ions, the molecule absorbs sufficient
energy and undergo fragmentation.

B+ + neutral
A+

C+ + neutral

Decompose to produce new ions

D+ + neutral

3. Separation of ions:
 The mixture of ions are separated according to the mass charge
ratio in the analyzer and then recorded.
 The record is known as the mass spectrum. It is the record of
abundance of each ion against its m/z value.

4. Mass spectrum:
 The mass spectrum is a plot of ion current intensity (ion
abundance) versus m/z value.
 The most abundant peak will give the tallest peak of the mass
spectra. This peak is known as the base peak and its mass
arbitrarily assigned a value of 100%. The heaviest peak is the
molecular ion peak and its mass will give the mass of the
molecule.

C+

B+
Relative absorbance ↑ D
A+

m/z value →

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

Isotope peak:
Isotopes present in the molecule may generate additional peak. Due to the
occurrence of isotopes we also observe M+1, M+2, M+3 etc peaks. The
relative abundances of these isotopic peaks are proportional to the
13
abundance of the isotope in nature (e.g. the natural abundance of C is
1.1% of the 12C atoms. For an ion containing n number of carbon atoms, the
abundance of isotope peak is nX1.1% of the 12C containing peak.
M+1 peaks are made by − 13C, 2H, 15N, 33S

M+2 peaks are made by − 18O, 34S, 37Cl, 81Br)

Base peak
M+ peak

Relative abundance ↑

M +1

M+2

m/z ratio →

M+1 and M+2 peak in benzene:


Benzene shows molecular ion peak at m/z value 78 due to C6H6. It will

also show M+1 peak at m/z 79 due to 13CC5H6+ or, C6H5D+.


13
M+2 peak will also show at m/z 80 due to CC5H5D+ or, 13C2C4H6+ or,

C6H4D2+.

The relative abundances of this isotopic can be used to determine


molecular formula.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

Atomic weight and natural abundances of some isotope −


Isotope Atomic Natural
weight abundance (%)
1
H 1.0078 99.985
2
H 2.014 0.015
12
C 12.000 98.9
13
C 13.003 1.1
14
N 14.003 99.64
15
N 15.0001 0.36
16
O 15.9949 99.8
17
O 16.999 0.04
18
O 17.999 0.2

Ionization method:
In ionization method compound are divided into 2 groups −
a. Ionization of volatile materials
b. Ionization of nonvolatile materials

a. Ionization of volatile materials −


Two methods are commonly used to produce ions from thermally
volatile compound ---
1. Electron impact ionization (IE)
2. Chemical ionization (CI)

1. Electron impact ionization:


• A direct probe tip is used near to a heated filament which
provides electron and is heated in the ionization chamber
causing vapor from the sample.

(Handle) (metal sheet) (Ceramic tip with sample)

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

• Electrons are accelerated from the hot filament to an anode, usually


through a potential difference of about 70ev.
• A 70ev electron has sufficient energy not only to ionize an organic
molecule but also to cause extensive fragmentation.
• Molecules are ionized due to bombardment with high-energy electrons
by removal of an electron. The product is cation radical.

M + e = M+ + 2e

2. Chemical ionization (CI):


• In this technique a reagent gas (methane, isobutane or ammonia) is
allowed to pass into the ion chamber at low pressure.
• The gas is ionized by using electron impact, which can then undergo
ion molecule reaction.

+
CH 4 + e → CH 4 • + 2e
+
CH 4 • + CH 4 → CH 5 + + CH 3 •

+
If the sample molecules are volatilized into mixture of ions, CH 5 act
as a strong acid and protonates the sample.

M + CH 5 + → MH + + CH 4

Thus in positive ion CI-spectra, the observed m/z value is one unit greater
than the true molecular weight.

 In CI-technique, negative ion CI-spectra may occur for molecules with


electron accepting properties like trifluoroacetates, quinones and nitro
compounds.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

b. Ionization of nonvolatile materials −


 molecules have low molecular weight but have numerous polar
functional groups, or
 have high molecular weight; usually don’t pass into the gas phase
at high temperature and at low pressure.

1. Field desorption:
• Here, the probe tip is replaced by a thin wire on which sharp
needles have been grown.
• The wire is supported between two posts on the probe.
• A solution of small amount of a sample is deposited on the wire.
• In the mass spectrometry the wire is maintained at +8kv and can
be heated and this can cause the discharge of an electron from
the sample into the metal. Thus positive ions (M+) are created.
In this way molecules are thrown into the gas phase as a
positive molecular ion without thermal decomposition.

Cathode slit
+8KV +
wire (+8KV) +
+
Probe +

Needle Ionized molecule

Fig: Field desorption technique

2. Desorption ionization by particles or radiation:


I. Laser desorption (LD)
II. Fast atom bombardment (FAB)
III. Californium plasma desorption
IV. Secondary ion mass spectrometry (SIMS)

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

 based upon giving a large pulse of energy to the sample


 here intermolecular bonds are broken and the sample is desorped
from its environment into the gas phase within 10-12 sec.
 so thermal decomposition doesn’t occur.

 Laser desorption →
 In this technique, the sample is bombarded with short duration,
intense pulses of laser light.
 Efficient and controllable energy transfer to the sample requires
resonant absorption of the molecule at the laser wavelength.
 Therefore, lasers emitting either in the UV or IR are employed.
 Laser pulses are applied for 1-100ns.
 One disadvantage is some thermo labile compounds may be
degraded with the laser beam resulting in a spectrum of
fragment ions. To overcome this problem, a matrix is used and
the technique is known as matrix assisted laser desorption
ionization technique (MALDI).

In MALDI, a low concentration of the sample is embedded either in a


liquid or a solid matrix (molar ratio 1:100 to 1:50000) which is
selected to absorb strongly the laser light. In this way, the energy is
transferred indirectly to the sample, and in a controlled manner
which avoids sample decomposition.
 Used in conjunction with a suitable method for ion analysis,
MALDI can give approximate molecular weight determinations
for biomolecules, even in the range 100000 – 200000 Daltons.

Some common MALDI matrices:

Matrix Application
Peptides, protein, lipids,
2,5-dihydroxy benzoic acid (DHB)
oligosaccharides
3,5-dimethoxy-4-hydroxy cinnamic
Peptides, protein, glycoproteins
acid (Sinapinic acid)
α-cyano-4-hydroxy cinnamic acid Peptides, protein, lipids,
(CHCA) oligonucleotides

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

 FAB →
 Here the energy is provided by a beam of neutral atom. The
sample is dissolved in a matrix of low volatility. A few µgm of
the sample are dissolved in a few µl of glycerol as matrix and a
beam of fast xenon atoms bombards the solution.

Bombarding atom beam

Probe sample ion Mass analyzer


Sample in matrix

 This fast xenon atoms are prepared by accelerating xenon ions


and then neutralizing these ions by charge exchanger at a low
pressure.

Xe+ + Xe → Xe + Xe+

Another matrixes used in FAB are ~


 Thioglycerol : Diglycerol (1:1)
 Tetracol
 Teracol
 Glycerol

 Californium plasma desorption →


 Here the sample to be analyzed is deposited on a thin metal foil
(nickel).
 Spontaneous fission of the radioactive Californium nucleus
(252Cf) occurs, and each fission event gives rise to two fragments
travelling in opposite directions. A typical pair of fission
fragments are 142
18 Ba + and 106
22 Te + of high kinetic energy.
 When such a high energy fission fragment passes through the
sample foil, produce a high temperature of 10000K.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100, Bangladesh

 Consequently the molecules in this plasma zone are desorbed


from the foil with the production of both positive and negative
ions. These ions are then accelerated out of the source into the
analyzer system.

252
Cf fission + −
or
Sample
Ni foil (10–3 mm)

 Californium plasma desorption technique produces better


molecular peak for molecules having molecular weight between
10000 – 20000 Dalton.

Different technique for analyzing ions in a mass spectrometer:


a. Magnetic sector
b. Time of flight
c. Quadrupole
d. Ion cyclotron resonance
e. Ion trap

Magnetic sector analyzer/mass analyzer:


 The ions may be separated according to their mass to charge ratio
(m/z) using a magnetic field.
 Here the ions of larger mass are deflected less than the ions of
smaller mass according to the equation –

Where,
m H 2r 2 r = radius of circular of path in which ion is
= traveling
z 2v H = magnetic field strength
V= potential difference of ion.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

The equation clarifies that by varying the magnetic field strength or


accelerating potential, the ions of all m/z value can be successively
allowed to pass through the detector slit & mass spectrum recorded.

Time of flight mass analyzer:


 TOF mass analyzer separates or resolves the ion beam by measuring
the flight time of the ions. The technique requires that all the ions
produced in the ion source should leave at the same time.
 The ions are accelerated by a potential difference and then allowed
to pass into the filed free region.
Since all the singly charged ions will acquire the same kinetic
energy, the largest mass will have the lowest velocity and the longest
time of flight over a given distance. Mathematically,

where
m 1 t = time of flight
t=L × L = distance in which a ion travel
e 2V m = mass of the ion
e = charge of the ion
V = velocity in which the ion travel

It is quicker than any other mass analyzer and applicable for all
masses.

Quadrupole:
 Here four parallel rods arranged symmetrically around an ion flight
path and a direct current and a radio frequency are applied to the
rods resulting an oscillating electrostatic field.
 The ions when pass through the region, will acquire an oscillation in
the electrostatic field. The ions of incorrect m/z ratio undergo an
unstable oscillation and strikes one of the rods.
 Ions of correct m/z ratio undergo stable oscillation of constant
amplitude and pass through analyzer to reach the recorder.

Quadrupole analyzer is a relatively compact instrument and


inexpensive.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Low-resolution mass spectrometer (LRMS):


LRMS employs a single stage analyzer. It resolves only integral masses
and it can’t differentiate the molecules e.g. CO (28), CH2=CH2 (28), N2
(28). As all have the same integral mass 28. since it can’t give exact
masses molecular formula can’t be determined.

High-resolution mass spectrometer (HRMS):


HRMS employs multiple stage analyzer such as magnetic and
quadropole sectors linked in series. The accuracy of these types of
instruments enables the distinction between different isotopes such as
13 12
C vs. C. The high-resolution data are obtained at an accuracy of
0.0001 amu (atomic mass unit) and consequently this permits a
distinction between species of the same mass unit such as - CO (28),
CH2=CH2 (28), N2 (28). Therefore data from HRMS are essential for
unambiguous determination of molecular data.

 Double focusing mass spectrometer are used to obtain high


resolution in which the beam ions are pass through an electric
field region before entering the magnetic field.
 In a single focusing mass spectrometer, there is a lack of
uniformity of ion energies that is all ions do not have precisely
same velocity. The result is peak broadening and low to
moderate resolution.

Electrospray ionization (ESI):


‘Electrospray’ is applied to the small flow of liquid (1-10µl/min) from a
capillary needle when a potential difference of 3-6kV is typically
applied between the end of the capillary and a cylindrical electrode
located 0.3-2 cm away.
 Under these circumstances, the liquid leaving the capillary does
not leave as a drop, but rather as a spray.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

 The spray consists of highly charged liquid droplets, and these


droplets may be positively or negatively charged depending on
the sign of the voltage applied to the capillary.
 If the liquid spraying from the capillary contains sample
molecules, then a molecular ion of these sample molecules can
be obtained by evaporation of the solvent.

ESI is an excellent technique for the production of molecular ions from


large polar molecules, and it will be seen subsequently that, since it
frequently produces multiply charged ions, it is a very powerful tool in
the analysis of biopolymers. This is especially true since the method
can be conveniently used to analyze directly the effluent from an HPLC
column.

Gas chromatography-mass spectrometry (GC/MS)


 The separation and detection of components from a mixture of
organic compound is readily achievable by gas chromatography.
Furthermore, limited characterization of unknown components is
often possible from retention times appropriate to the particular
column used.
 Mass spectrometry, because of its high sensitivity and fast scan
speeds, is the technique most suited to provide definite structural
information from the small quantities of material eluted from a gas
chromatograph.
 The association of the two techniques provided a powerful means of
structure identification for the components of natural and synthetic
organic mixtures even though the components may be present in
nanogram quantities and eluted over periods of only a few seconds.

The interface between the GC and the MS is a jet separator.

Such a combination is useful as an aid in determining the structures


and chiralities of amino acids. The amino acids are first derivatized as

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

follows: by treatment with trifluoroacetic anhydride in the first step


and by isopropanol / HCl in the second (to render them volatile):

NH2CH(R)COOH → CF3CONHCH(R)COOH → CF3CONHCH(R)COOCH(CH3)2

Since trifluoroacetyl is a good electron capture group, the mass


spectra are determined in the negative ion mode. The mixture of
derivatized amino acids (frequently from 6N HCl hydrolysis) is simply
injected on to a chiral GC column, where the retention times are not
only dependent on structure but also on absolute configuration of the
amino acids. Thus separation, molecular weight, and chirality can all
be determined in one experiment.

Liquid chromatography-mass spectrometer (LC/MS)


• HPLC is a powerful method for the separation of complex
mixtures, especially when many of the components may have
similar polarities. In reverse-phase HPLC, the column substrate is
such that starting with an aqueous solution of a mixture of polar
components; the most polar components are eluated first. The
later-eluated hydrophobic components are often encouraged to
leave the column by gradually increasing the concentration of
acetonitrile (CH3CN) in the otherwise aqueous developing
solvent.
• If the mass spectrum of each component can be recorded as it
eluates from the HPLC column, quick characterization of the
components is greatly facilitated.

Tandem mass spectrometry (MS/MS):


• Tandem mass spectrometer uses two mass spectrometers is
tandem.
• It has a great potential value in the structure elucidation of
organic compounds.

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

• In this technique, a compound to be analyzed is subjected to


ionization and fragmentation. The mixtures of ions are then
separated in the first mass analyzer.
• From the mixture of ions, a specified ion is selected for the
second mass analyzer. The magnetic field is set to pass only the
selected ion through a slit into a collision chamber. This chamber
contains a high energy reagent gas like helium (He) or argon (Ar)
with which the ions collide. As a result, mixtures of ions are
produced. The process is known as collision activated
decomposition (CAD). The ions are then analyzed in the second
mass analyzer.

Example –
Penazitidine A is a heterocyclic compound with a long chain. It has a
methyl group on the side chain but the position was not established by
various technique (NMR & even by 2D NMR). But MS-MS can determine
the position of methyl group on the structure.

The ion at m/z 296 was selected. It was allowed to pass into the
collision chamber where it is subjected to CAD. The mixtures of ions
produced are analyzed in the 2nd mass analyzer. The intense peak at
m/z 182, m/z 210 indicates the position of methyl group at C12 of the
side chain.

Fragmentation patterns:
For most classes of compounds, the mode of fragmentation is
somewhat characteristic. The most common mode of fragmentation
involves the cleavage of one bond. In this process the odd-electron
molecular ion yields an odd-electron neutral fragment (a radical) and
an even electron fragment ion (carbonium type)

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Fragmentation via the cleavage of one bond:


+
[R − CH ]3

→ R + + • CH 3
+
[R − X ] → •
R+ + X • [odd elecetron radical]
[old electron molecular ion] [evene electron fragment ion]

where, X = halogen, OR, SR or NR2, R = H, alkyl or aryl

Fragmentation via the cleavage of two bonds:

+ +
[RCH − CHR ] / •
→ [RCH = CHR / ]• + H 2O

+ +
[RCH − CH 2 − O − C − CH3 ] →

[RCH = CH ] 2

+ HO − C − CH3
(odd electronmolecularion) (odd electronfragmention) (evenelectronneutralfragment)

In addition to this process, fragmentation process involving


rearrangements, migrations of groups and secondary fragmentations of
fragment ions are also possible.

Fragmentation patterns of different classes of compound:

Name of M+ peak Fragmentation


compound
Alkane − M+ peak is observed but less C-C bonds leading to the formation of 20 & 30
i) Branched intense than straight chain carbonium ions which are more stable than 10.
chain hydro- compound As a result the M+ ion will become less intense
carbon. and undergo further fragmentation.
e.g. +
• Cleavage of a C-C bond yields an
[CH 3 − CH − CH 3 ]∗
Isobutane isopropyl carbonium ion.
CH3 +
[CH 3 − CH ]

CH3-CH-CH3
CH3
0
2 carbonium ion, m/z =43

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Alkanes − Molecular ion peak observed at C-C bonds breaks resulting in a homologous
ii)Straight m/z = 58 series of fragmentation products. Primary
chain carbonium ion is formed.
compound + 1. Cleavage of C-1 to C-2 bond result in loss of
[CH 3 − CH 2 − CH 2 − CH 3 ]•
e.g. − Butane a Me-radical and formation of propyl
carbonium ion.
[CH 3 − CH 2 − CH 2 ]+ + [CH 3 ]• (m/z = 43)
43
2. Cleavage of c-2 to c-3 bond result in loss of a
Et-radical and formation of ethyl carbonium
29
ion
15 58 [CH 3 − CH 2 ]+ + [CH 3 − CH 2 ]• (m/z = 29)
3. Cleavage of c-3 to c-4 bond results in loss of
a propyl radical and formation of methyl
11 15 25 30 40 45 50 55 60 carbonium ion
(m/z) → [CH 3 ]+ + [CH 3 − CH 2 − CH 2 ]• (m/z = 15)
Fig: Mass spectrum of Butane

Alkenes − Distinction M+ peak Alkene isomers show nearly identical mass spectra.
E.g. − 1-butene So double bond can’t be located. Also cis & trans
& 2-butene • 1-butene and 2-butene can’t be differentiated.
give molecular peak at
CH2=CH-CH2- m/z =56. +

CH3
[R − CH 2 − CH = CH 2 ] → R + CH2 − CH = CH 2
• • +

• Both produce allyl


1-butene carbonium ion at m/z =
+
41. [CH 2 = CH − CH 2 − CH 2 − R ]• → R • +
[CH 2 CH 2 − CH = CH 2 ↔ CH 2 = CHCH 2 − CH 2

+
[CH 2 = CH− CH2 − CH3 ]• → CH3 + CH2 = CH− CH2
• +

(m/z = 41)
+
Alkynes − • Molecular ion peak is
[H − C ≡ C − CH 2 − R] → R • + •

E.g.− Propyne rather intense. +

• Give molecular ion peak [H − C ≡ C − C H 2 ↔ HC = C = C H 2 ]

at m/z = 40 +
 + +

[HC ≡ C − CH 3 ]• →  HC = C C H 2 ↔ H C = C = CH 2 
 
(Propyne) (m/z = 39)

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Aromatic • Show distinct and intense Fragmentation occurs at benzylic position not at
hydrocarbon molecular ion peak. phenolic position.
i) containing
alkyl group
E.g. • Not so intense Fragmentation occurs at benzylic position. Loss of
Tolune • Shows very intense hydrogen gives a strong peak at m/z = 91
molecular ion peak at m/z
CH3 = 92
+ +
CH2
+
CH3
+
C7H7
Benzyl carbonium ion Tropylium ion (m/z = 91)
(m/z = 91)

Ethyl benzene Molecular ion peak gives at Fragmentation occurs at benzylic position. Loss of
m/z = 106 hydrogen gives a strong peak at m/z = 91
CH2-CH3 +
CH2 +
+
CH2-CH3 + CH3

Propyl Molecular ion peak at m/z = Fragmentation occurs at benzylic position. Loss of
benzene 120 hydrogen gives a strong peak at m/z = 91
+ +
CH2
CH2-CH2-CH3 +
CH2-CH2-CH3 + CH2-CH3

Alcohol Intensity of molecular ion • Loss of an alkyl group. C-C bond broken.

1º & 2º peak is usually low. CH3 − CH2 − C H 2 + CH2 = OH+ m/z=31
+
3º [CH3 − CH2 − CH2 − CH2 − OH] •
(1 − butanol) •
CH3 − C H 2 + CH3 - CH = OH+ m/z=45
+
[CH3 − CH2 − CH − OH]• •
CH3 C H 3 + (CH3 ) 2 - C = OH+ m/z=45
(2 − butanol)

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

+ • Dehydration: the importance of dehydration


CH3 * increases as the chain length increases. The
CH3-C-OH dehydration is a 1,2 elimination of water.
CH3 General reaction -

(CH2)n (CH2)n

RCH CHR' + H2O


RCH CHR'
H OH

Example - (1-butanol)

(CH2)2
CH2
CH2 CH2 + H2O
CH2 CH2
H OH

• Simultaneous loss of water & alkene usually


which contains more than four carbon.
+
H H
RCH +
O
R C CH2 + CH2=CH2 + H2O
H2C CH2 H

C
H2

Ex − 1-butanol −
+
H
O
H +
H2C
H2C CH2 + CH2=CH2 + H2O
H2C CH2
m/z = 28
C
H2

Phenols − Exhibit intense molecular ion • Loss of CO


E.g.−Benzyl peak.
alcohol 108 OH H H
(M) - CO
m/z = 66
79 107
77
+
OH H H
CH2OH +
- CO +
+ H C6H5 + H2

m/z=107 m/z=79 m/z = 77

Fig: Mass spectrum of


benzyl alcohol

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Ethers − Weak molecular ion peak. • Loss of alkyl group/cleavage of C−C bond to
the α-carbon
α + +
[R − C H 2 OR' ] → CH 2 = O R'+ R •

• Cleavage of C−O bond


α + +
[R − C H 2 OR' ]• → C H 2 − R + R' O •

Aromatic Molecular ion peak is • Loss of alkyl group


ethers − observed + +
[C 6 H 5 OR]• → C 6 H 5 O + R •
E.g. − Anisole
• Loss of alkoxy group
+
+
[C 6 H 5 OR]• → C 6 H 5 + OR •
• Example − Anisole
+ +

[C 6 H 5 - O - CH 3 ]• → C 6 H 5 O + CH 3
+
+ •
[C 6 H 5 - O - CH 3 ]• → C 6 H 5 + OCH 3

Aliphatic Molecular ion peak is • α-Cleavage: cleavage occurs in one of the two
aldehyde − observed/ weak bonds to the carbonyl group.
O
+
R C H R C O + H *

O
+
R C H H C O + R *

R C H H C O * +R+

• β-Cleavage: cleavage occurs in β- carbon.


R − CH 2 − CHO → R + + CH 2 = CHO •
• Mclafferty rearrangement: when alkyl group
attached to the carbonyl carbon is large, a type
of rearrangement called Mclafferty
rearrangement occurs.
+
H +
R1 CH O R 1 -C H
C H2
+
R2 CH C H -O H
C R 2C H
CH2 H m /z=4 4

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

e.g. mass spectrum of Butyraldehyde: molecular ion peak at m/z=72.


+ + +
O CH3CH2CH2=O + H m/z=71

CH3-CH2-CH2-C-H +
H-C O + CH3CH2CH2 m/z=29
α-Cleavage:
+
O
+
CH3-CH2-CH2-C-H CH3CH2 + CH2=CHO m/z=29
β - Cleavage:
Mclafferty rearrangement:
H +
CH2 +
O +
CH2 CH2 + CH2 CH -OH m/z=44
CH2
CH
CH2

Aromatic • Intense molecular ion • Loss of hydrogen show molecular peak at


aldehyde − peak. m/z = 105
• Benzaldehyde show
e.g. +
molecular peak at m/z = O +
Benzaldehyde 106 C O
C H +H
O

C H 106(M+) m/z=105
105
77 • Loss of CHO group may lead to C6H5+
which show molecular peak at m/z = 77
C6H5+ C6H5C≡O+
Fig: mass spectrum of
Benzaldehyde
Aromatic Intensed molecular ion peak • loss of alkyl radical giving ArCO+ ( α -
ketones − Cleavage)
+
O +
C O +
C R +R*

m/z=105 m/z=77

• loss of COR giving Ar+ (Mclafferty


rearrangement)
H R H +
+
O CH O +
C O
CH2 C
C
CH2
CH2 m/z=105
m/z=120
+ CHR CH2

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M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Acyclic • Intense molecular ion 1. α-Cleavage


Ketones − peak. +
O + *
R -C O + R'
R - -- C -- - R '

+
O
+ *
R '- C O + R
R --- C ---R '

2. Mclafferty rearrangement:
+
+
H OH
R 1C H R 1 CH
O + C CH2
R 2C H R 2C H
CR3 R3
CH2

e.g.− +
O
+
2-butanone CH3-C O + CH3CH2
CH3-C-CH2CH3
m/z=43
larger gr. will be predominant

+
O
+
CH3CH2-C O + CH3
CH3--C-CH2CH3
m/z=57

the peak at m/z =43 is more intense than the peak at


m/z =57
Aliphatic May be very weak or even β - Cleavage:
amines − absent. + +
R-CH2-NH2 * R * + CH2 NH2
m/z=30

Aromatic Intense molecular ion peak Loss of hydrogen atom.


amines − + + +
NH2 + NH * H H H *
* *
H* *
HCN + H*

m/z=92 m/z=66 m/z=65

Aromatic Intensed molecular ion peak Loss of OH to form C6H5CO+ ion (m/z = 105)
Carboxylic followed by loss of CO to form the C6H5+ ion
acids − O + +
+
* C O * +
C OH

m/z=105 m/z=77
+ OH *

21
M. Kaisarul Islam, Lecturer, Department of Pharmacy, Jagannath University, Dhaka-1100,
Bangladesh

Aromatic Weak molecular ion peak α - Cleavage gives to the formation of C6H5CO+
esters − 105 O + +
C O +
*
E.g. − Methyl C OCH3
benzoate. 77 m/z=105 m/z=77
136 + OCH3*
+
(M )

Fig: mass spectrum of


methyl benzoate
Esters − Weak but noticeable ion • α-Cleavage: loss of the alkoxy group to form
peak corresponding acylium ion RCO+
O +
*

+
R C-OR R *

+ O C-OR'
'

O + O
*

+
R COR' R + *
C-OR'

+
O
*
+
R-C OR' R C O + *
OR'

• β-Cleavage: Mclafferty rearrangement


H +
OH +
R1CH * R1CH
O *
+ C CH2
R2CH
C R2CH
CH2 OR' OR'

22

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