Relative abundance of isotope & their contribution

to characteristics peaks & Mass spectrum.

Presented by : Eknath A. Hirve
M.Pharm 1st pharmaceutics
Submitted to : Dr. G. S. Gadaginamath
Modern Pharmaceutical Analysis
Department of pharmaceutics
SET’s college of pharmacy
DHARWAD
Content-
 1.Principle
 2.Mass spectrum
 3.Presentation of mass spectrum
 4.Component of mass spectra
 5.Abundance of the isotope
 6.Contribution of isotope to characteristic peaks
 7.Reference


 Principle:
 Organic molecule are bombarded with electrons and converted to highly
energetic positively charged ions (molecular ions or parent ion),which can
break up into smaller ions (fragment ions or daughter ions).,the loss of an
electron from a molecule leads to a radical cation (M M•+ ).
 The molecular ion M•+ commonly decomposes to a pair of fragments, which may be
either radical plus or small molecule plus a radical cation.



The molecular ions, the fragment ions and the fragment

radical ions are separated by deflection in a variable
magnetic field according to their mass and charge, and
generate a current (the ion current) at the collector in
proportion to their relative abundances.

 Mass spectrometer


 ☻ Mass spectrum:-
The mass spectrum is a plot of relative abundance of the
positively charged ions and fragments against their mass to
charge ratio (m/z).
The X-axis indicates mass to charge ratio (m/z).

The left ordinance of Y-axis indicates the relative abundance

(intensity).




q A mass spectrum shows-

vThe mass of the molecule from molecular peak.
vThe mass of the fragments formed from the molecule.
vrelative abundances of all the ions or fragments.
üRelative abundance, which related to the number of times an
ion of that m/z ratio, strikes the detector.
üRelative abundance is determined by measuring the height,
because peak height is proportional to no. of ions produced.
Larger the peak ,more abundant the ion producing it.




Presentation of the spectrum:
v Bar graph
v Tabular form
1.Bar graph:


 Ex. Dopamine
 The five traces that are obtained with a multiple
galvanometer system. Because a spectrum of this sort is
difficult to use, usually this information is converted into a
form of a bar graph.
Bar graph: (dopamine)
 Advantages:
üNumber of peaks which are not of interest thus showing only
the intense peak.
üFragmentation pathway can be directly suggested & contains
less data.
 Disadvantages:
üInability to show the less intense ion peak.
üInability to show the less intense isotopic peak.



2.Tabular form:


Commonly mass spectral result may also be presented in



tabular form.
Advantages:


üClearly visible, all information in a single page.

üLess intense peaks can be clearly represented.
 Bar graph (pentane)
Tabular form
Components of mass spectra:

Molecular ion and base peak-

ØThe ions formed from a molecule by the loss of one electron

from molecule.
ØWhen the vaporized organic sample passes into the ionization
chamber of a mass spectrometer, it is bombarded by stream
of electrons.
ØThese electrons have a high enough energy to knock an
electron off an organic molecule to form a positive ion. This
ion is called the molecular ion.
Ø
ØThe molecular ion is often given the symbol M+ or M•+ .
ØThe mass spectrum of pentane, the line produced by the
heaviest ion passing through the machine (at m/z =72) is
due to the molecular ion.
ØThe tallest line in this spectra (at m/z=43) is called the base
peak.
ØThe base peak is tallest peak because it represent the
commonest fragment ion to be formed.


The species due to the final signal is known as the molecular

ion and is usually corresponds to the molecular mass of the
compound.

Ex. ethyl benzene (mw.106)
C6H5CH2CH3 + e- C6H5CH2CH3•+ + 2e-
 M+1 peak:
 If we have a complete mass
spectrum, we will find a small line
1 m/z unit to the right of the main
molecular ion peak. This small peak
is called the M+1 peak.

 THE MOLECULAR ION
 The small peak (M+1) at 115 due to the natural abundance (about 1%) of carbon-13.
The height of this peak relative to that for the molecular ion depends on the number of
carbon atoms in the molecule. The more carbons present, the larger the M+1 peak.
100


80
Abundance %

60
40

.
10 20 30 40 50 60 70 80
20

90 100 110 120 130 140

m/z

 Carbon -13 isotope:

▪ The M+1 peak is caused by the presence 13 C isotope in the



molecule.
 ▪ Methane(CH4) approximately 1 in every 100 of these molecule
will contain C13 rather than more common C12.
 ▪ That means that 1 in every 100 of the molecule will have a mass
of 17(13+4) rather than 16(12+4).
 ▪ The mass spectrum will therefore have a line corresponding to the
molecular ion (13 CH4)+ as well as (12 CH4).+






Isotope:- meaning at the same place, that all isotope of an

element are located at the same place on the periodic table.
Isotope is an atom which contains a different number of
neutrons in its nucleus .
Means different isotope of an element will have different
masses.
Since both proton & neutrons contribute equally to the mass
of an atom.
The sum of the protons & neutrons is the mass number.
 Ex. 3He(2 p - 1 n), 4He (2p – 2 n).
Isotope have same chemical properties but different weight.

Abundance of the isotope:-
▬ Not all isotope are equally abundant in nature.
▬ Ex. Naturally occurring isotope of Hydrogen (Hydrogen-2 is
the only common isotope which has its own name, called
Deuterium).
Compare these abundances of hydrogen to those for some other
elements, notice that the atomic mass is close to the sum of the
number of the neutrons and protons, but not exactly the same.
Natural abundances of common elements and their isotopes:-


Sr. no. Elements Relative

abundance %
1 Carbon 12 100
2 Carbon 13 1.08
3 Nitrogen 14 100
4 Nitrogen 15 0.38
5 Oxygen 16 100
6 Oxygen 17 0.04
7 Oxygen 18 0.20
8 silicone 28 100
9 Silicone 29 5.10
10 Silicone 30 3.35
11 Sulfur 32 100
12 Sulfur 33 0.78
13 Chlorine 35 100
14 Chlorine 37 32.5


Contribution of isotope to characteristic peak:-
Ethane (C2H6) has a mw.30 when it contain most common isotope
of carbon and hydrogen.
Occasionally sample of ethane yields a molecule in which one of
the carbon atom is heavy isotope of carbon C13.
This mol. Appear mass spectra at 31.
The relative abundance of C13 in nature is 1.08%.
There are two carbon atom in in ethane, a molecule Of mass 31
will turn up (2x1.08) or 2.16% of the molecular ion peak.
This mass 31 peak is called as m+1 peak.



If a deuterium atom, H2 replaced one of the hydrogen atom in

ethane, the molecule also have mass of 31.
The natural abundance of deuterium is 0.016%.
The intensity of the m+1 peak would be (6x0.016) or 0.096%
of the intensity of the molecular ion peak.
C-13- (2x1.08) 0r 2.16 intensity of the m+1 peak.




How the intensities of the m+1 peak----

Consider two molecule of mass 42, propane (C3H6) &

diazomethane (CH2N2).
PROPANE- the intensity of the M+1 peak should be (3x1.08)
+ (6x0.016)=3.34 % .
DIAZOMETHANE- the natural abundance of N15 isotope of
nitrogen is 0.38 %. Relative intensity of the M+1 peak to be
1.08+ (2 x 0.016) + (2 x 0.38)=1.87 % of the intensity of the
molecular ion peak.

 REFERENCE
Jag Mohan., Organic spectroscopy principal and
application., II edition, Narosa prakashan.
Skoog. Holler. Crouch., Instrumental analysis., India edition.
Gurdeep R. Chatwal, Sham K. Anand., Instrumental methods
of chemical analysis., Himalaya publication house.
Y.R Sharma., Elementary organic spectroscopy., S. Chand.
Dr. A.V. Kasture, Dr. K.R. Mahadik., pharmaceutical
analysis, vol -II, Nirali prakashan.


Questions:-


Explain the general importance of M, M+1, peaks.

Outline of the principle of mass spectrometry.