INTERPRETATION OF

MASS SPECTRA

Renjitha J R
Assistant Professor
Mount Zion College Of Pharmaceutical
Sciences And Research , Adoor
Mass spectrum

Mass spectrum is a plot of mass to charge ratio of ions (x axis) against

relative abundance of ions (y axis) .

As most of the ions formed are singly positively charged peak position
denotes the mass of the ion.

Peak height denotes the relative abundances of the ions, where relative
abundance is the no: of times a molecule of a particular m/z value
strikes the detector.
Various Peaks In Mass Spectra

Molecular ion peak or parent peak or mass peak: peak at the

highest m/z value.
Base peak : most abundant peak in the spectra.

Isotope peaks:
M+1 peak : formed by 13C, 2H,15N, 33S
Relative intensity of M+1/1.1 gives no of carbon atoms

M+2 peak : formed by 18O, 34S, 37Cl, 81Br

Relative intensity indicates presence of Cl, S, Br in a compound
A Typical Mass Spectrum
 Rules For Interpreting Mass Spectra

Isotope abundance
Nitrogen rule
Hydrogen rule
Index of hydrogen deficiency
Ring rule
Rule of thirteen
Fragmentation pattern
Isotope Abundance:

Isotopes:
Atoms having same atomic number but different mass number.

Eg: Carbon - 12C ,13C

Hydrogen - 1H, 2H, 3H
Oxygen - 16O, 17O, 18O
Chlorine - 35Cl ,37Cl
Isotopic abundance:

Mono isotopic elements: F , P , I

M+1 isotope peak - formed by isotopes of elements which differ

by 1 mass unit.
Eg: C , H , N

M+2 isotope peak - formed by isotopes of elements which differ

by 2 mass units.
Eg: Cl , S , O
 M contributors M+1 contributors M+2 contributors

Relative Relative Relative

Isotope Abundanc Isotope Abundanc Isotope Abundanc
e e e
1
H 100 2
H 0.016%
Relative Isotope Abundances of Common Elements
12
C 100 13
C 1.11
14
N 100 15
N 0.38
16
O 100 17
O 0.04 18
O 0.20
19
F 100
32
S 100 33
S 0.78 34
S 4.40
35
Cl 100 37
Cl 32.5
79
Br 100 81
Br 98.0
127
I 100
When bromine or chlorine is present in a compound, the M+2 peak
becomes very significant.

This is due to the fact that, for bromine, two isotopes (79Br and 81Br) are
present in a 1:1 ratio in naturally occurring substances.

For chlorine, two isotopes (35Cl and 37Cl) are present in a 3:1 ratio in
naturally occurring substances.

Molecular ion peak is assigned an abundance of 100 %.

Abundance of other isotopic peaks are normalized with respect to the

molecular ion peak.
 M: m/z = 84
Isotope ratio:
From isotope ratio CN4O
C2N2O2
Number of % peak height C2H2N3O
atoms = % abundance C2H4N4
Molecular ion peak = 84 C5H8O

(M+1) peak = 5.57%

(M+2) peak = 0.3%
Number of carbon atoms = 5.57 = 5.06
1.1
Nitrogen Rule:

A molecule of even numbered molecular weight may contain

either no nitrogen or an even number of nitrogen atoms and a
molecule of odd numbered molecular weight contains odd
number of nitrogen atoms.
 CH3

O2N NO2

Example :
NO2

formula: NH3 H2NNH2

m/z : 17 32 227

odd even odd

Hydrogen Rule:

Maximum no of hydrogen and halogen atoms that can be

present in a molecule is given by equation:
H = 2C + N + 2

Applicable for molecules containing C, H, O, N, Cl, Br ,F, I.

Molecule may contain lesser no of atoms than predicted by
this rule.
Example :

max no: of H=

 
max no: of H reduced by 1

max no: of H reduced by 2

Hydrogen Deficiency Index:

Calculated from molecular formula

Also called number of sites (or degrees) of unsaturation or
double bond equivalent {DBE}.

HDI = no of rings + no of double bonds + twice no of triple

bonds
or
HDI = number of carbon atoms – number of hydrogen atoms/2
–number of halogen atoms /2 + number of nitrogen atoms/2 +
1.
Example:

CcHhNnOoXx
HDI = c - h/2 – x/2 + n/2 +1

C7H7NO :

HDI = 7 - 3.5 - 0+ 0.5 + 1 = 5

If HDI = 1, compound shows one pie bond or one ring.

If HDI = 2, compound shows two rings or double bonds or one of

each or one triple bond.

If HDI = 4, possibility for benzene nucleus{one ring and three

double bonds}.
Ring Rule:

To determine any unsaturation in the compound.

CcHhNnOo

R = c+ 1 + n – h
2
Example: for C6H6 :

c=6, h=6, n=0

Then R=6+1-3 =4
Rule Of Thirteen
Used to calculate no of carbon and hydrogen atoms in alkanes.
Divide molecular weight by 13.
Numerator gives no of carbon atoms.
Numerator +(13* remainder) gives no of hydrogen atoms

 
 
 
 
• Example:
If molecular weight is 400. Divide 400 by 13 which is
30.7692 .

30 is the number of carbon atoms.

  Multiply the remainder (.7692) with 13 to get 10.

Add number 10, to the whole number 30, to get the number of
hydrogens, 40.

So the molecular formula is which equals 400.

 
Fragmentation Pattern
The relative abundance of various masses of ions in a spectra is
characteristic of a particular compound under specified conditions.
This pattern is known as fragmentation pattern.

 
Uniqueness of fragmentation pattern

Peaks differing in mass by 14 - straight chain hydrocarbon

Alcohols - strong peak at (M-18)

Primary alcohol – at m/z 31 due to OH+

Ion  Fragment Lost  Possible Inference 
M-1 H 
M-15 CH3
M-16 O, NH2 amide
M-17
M-18 Common Fragment Ions:
OH
H2O alcohol
M-19 F  fluoro compound
M-26 C2H2  aromatic
CO   ketone aromatic, ethyl ether, ethyl
N2   ester, n-propyl ketones,
M-28
McLafferty  
C2H4  
rearrangement
CHO   aldehyde  
M-29 C2H5  ethyl
M-30 CH2=NH2 primary amine
M-31 OCH3  methyl ester
M-32 CH3OH    methyl ester
M-35, 37 Cl chloro compound
C3H7   propyl  
M-43
CH3CO methyl ketone
M-45 CO2H    carboxylic acid
M-46 NO2  nitro aromatic
M-57 C4H9 butyl
M-60 CH3CO2H  acetate
M-79,81 Br  bromo compound.
Fragment ( m/z) Possible Group Possible Inference 
29  CHO+, C2H5+
30 CH2=NH2+ primary amine
31 CH2=OH+ primary alcohol
41 C3H5+ allyl group
C3H7+    propyl group  
43
CH3CO+   acyl group
50 C4H2+  aromatic
51 C4H3+ phenyl group
C4H9+    butyl group  
57
C2H5CO+ ethyl ketone, propionate ester 
58 CH2=C(OH)CH3+  methyl ketone (McL. Rear.)
CO2CH3+   methyl ester   
59
CH2=C(OH)NH2+ primary amide (McL. Rear.) 
 CH2=C(OH)OH+  carboxylic acid (McL. Rear.) 
60
C5H11+   pentyl group  
71
C3H7CO+ propyl ketone, butyrate ester  
74 CH2=C(OH)OCH3+ methyl ester (McL. Rear.)

C6H4+  mono or disubstituted

76
benzene
77 C6H5+ phenyl
85, 99 CcH2c+1+  benzyl group
91 C7H7+  alkyl group
105 C6H5CO+  benzoyl group
Steps To Interpret The Mass Spectra

1.Look for the molecular ion peak.

This peak will be the highest mass peak in the spectrum,
except for isotope peaks.

Nominal MW (meaning=rounded off) will be an even

number for compounds containing only C, H, O, S, Si and
even no: of Nitrogen or no Nitrogen.

Nominal MW will be an odd number if the compound

contains an odd number of N (1,3,...).
2.Try to calculate the molecular formula:

The isotope peaks can be very useful.

Example:
Carbon 12 has an isotope, carbon 13.  Their relative abundances
are 12C=100%, 13C=1.1%.  This means that for every 100 (12)C
atoms there are 1.1 (13)C atoms.

If a compound contains 6 carbons, then each atom has a 1.1%

abundance of (13)C.
if the molecular ion peak is 100%, then the isotope peak would be
6x1.1%=6.6%.
If the molecular ion peak is not 100% then you can calculate
the relative abundance of the isotope peak to the ion peak.
 
For example, if the molecular ion peak were 34% and the
isotope peak 2.3%:  (2.3/34)x100 = 6.8%.  6.8% is the
relative abundance of the isotope peak to the ion peak. 

Next, divide the relative abundance by the isotope

abundance:  6.8/1.1=6 carbons.
3.  Calculate the total number of rings plus double bonds:
either using the Ring rule or Hydrogen Deficiency
Index.
4.  Postulate the molecular structure consistent with peak
abundance and m/z of fragments.
Example:
m/z Molecular ion Relative abundance Conclusions
102 M 100 % Mass (lowest isotopes) = 102
Given information

Even number of nitrogens

103 M+1 4.50 %
4.50 / 1.1 = 4.09 approx.:4
carbons

104 M+2 0.68 % 0.68/0.2 = 3.4 approx. : 3 oxygen

< 4% so no S, Cl, or Br
Possible molecular formula are:

 
Applying hydrogen rule:

H= 2C+ N+1
= 2*4+ 1
=9
so the possible formula can be
𝐶 4 𝐻 4 𝑂3
 
Applying ring rule:
R= C + N/2 – H/2- X/2 +1
= 4- 4/2+1
=3
Mass Spectra Of
Some Chemical Classes
HEXANE

ALKANES
Alkenes characteristically
 show a strong molecular ion peak.
 cleave readily to form resonance-stabilized allylic cations.
ALKENES +
+
[CH2 =CHCH2 CH2 CH3 ] • CH2 =CHCH2 + • CH2 CH3
ALCOHOLS

Eg: Pentane-1-ol C5H12O

MW = 88.15
ETHER
Eg: Ethyl ether
KETONE
Eg : pentane-3-one
MW = 86.13 g/mole
ALDEHYDE
Eg: Hexanal
MW = 100.15888
CARBOXYLIC ACIDS
Eg: Hexanoic acid
MW = 116.1583 g/mole
Conclusion
Mass spectra can be used for determining the molecular
weight , molecular formula and even the structure of the
molecule.

Molecular ion peak and isotopic peaks are most useful for
the interpretation of a mass spectrum.

Mass spectrometry because of its high sensitivity and fast

scan speed , can be interfaced with a GC which provides
definite structural information from small quantities of
eluted materials.
References

1. Silverstein Spectrometric Identification Of Organic Compounds

Sixth Edition :Wiley&sons,Inc. Newyork1998
2. William Kemp Organic Spectroscopy Third Edition
ELBS:Hong Kong 1993
3.Gurdeep R Chatwal Instrumental Methods Of Chemical Analysis
Fifth Edition :Himalaya publishing house. Mumbai 2002.
4.Y.R.Sharma,Elementary Organic Spectroscopy,2006 Edition
5.Willard & Meritt instrumental methods of analysis, seventh
edition
Thank You