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Mass Spectrometry (MS)

Measures molecular weight


Sample vaporized and bombarded by electrons
that remove an electron, creating a cation-radical
Bonds in cation radicals begin to break (fragment)
Charge to mass ratio is measured

Mass Spectrometer

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Spektrometer massa

amplifier

Komponen dasar MS
1. Sistem injeksi sampel
2. Sumber pengionan
3. Analisator/pemisah massa
4. Sistem pompa vakum
5. Detektor
6. Penguat sinyal
7. Pencetak spektra

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The Mass Spectrum
Plot mass of ions (m/z) (x-axis) versus the
intensity of the signal (roughly corresponding
to the number of ions) (y-axis)
Tallest peak is base peak (100%)
Other peaks listed as the % of that peak
Peak that corresponds to the unfragmented
radical cation is parent peak or molecular
ion (M+)

MS Examples: Methane and


Propane
Methane produces a parent peak (m/z = 16) and
fragments of 15 and 14 (See Figure 1a)
The MS of propane is more complex (Figure 1b) since
the molecule can break down in several ways

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3-methyl-6-i-PropylCyclohex-2-ene-1-one

MW = 152

Isotopic cluster M, M+1, M+2

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Interpreting Mass Spectra
Molecular weight from the mass of the molecular
ion
Double-focusing instruments provide high-
resolution exact mass
0.0001 atomic mass units distinguishing specific
atoms
Example MW 72 is ambiguous: C5H12 and
C4H8O but:
C5H12 72.0939 amu exact mass C4H8O 72.0575 amu
exact mass
Result from fractional mass differences of atoms 16O
= 15.99491, 12C = 12.0000, 1H = 1.00783
Instruments include computation of formulas for
each peak

Natural Abundance of a few elements

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Molecular formula in
Molecular ion:
general

Even Mass : can contain C, H, O , halogen, even # N


Odd Mass : can contain C, H, O , halogen, odd # N

Fragmentation ions:
Even Mass : From even mass M+
comes from rearrangement or 2 bond breaking
Odd Mass : From even mass M+
comes from single bond breaking

even Mass : from Odd Mass M+


comes from single bond breaking

Molecular formula: RULE of 13


Consider CH unit 13 amu

If we divide the mass by 13 we can establish easily possible formula:


Example: Molecular ion => 152
152 / 13 = # carbons = 11
Mass = 12 * 11 = 132 therefore H = 152 132 = 20

Basic formula C11 H20


If Oxygen is present : mass = 16 remove CH4
If Nitrogen is present : mass = 14 remove CH2
If one oxygen : C11 H20 CH4 + O = C10 H16 O
If second oxygen : C10 H16 O CH4 + O = C9 H12 O2
If third oxygen : C9 H12 O2 CH4 + O = C8 H8 O3

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Molecular formula: RULE of 13
After establishing the basic formula with only Carbon/hydrogen,
Other element can be introduced by substracting the proper hydrocarbon
value
14N => CH2 16O => CH4
19F => CH7 1 H12 => C
28Si => C2H4 127I => C10H7
31P => C2H7 79Br => C6H7
32S 35Cl => C2H11
=> C2H8

Nitrogen
rule

Molecular ion => 26 Molecular ion => 27 odd!


26 / 13 = # carbons = 2 27 / 13 = # carbons = 2
Mass = 12 * 2 = 24 therefore H = 2 Basic formula C2 H3
Basic formula C2 H2 One Nitrogen C2 H3 - CH2 + N
H C C H H C N

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Nitrogen rule NH2
OH

Molecular ion => 100


100 / 13 = # carbons = 7
Molecular ion => 99 odd!
Mass = 12 * 7 = 84 therefore H = 16
Basic formula C7 H15
Basic formula C7 H16
1 Nitrogen: C7 H15 CH2 + N
1 Oxygen: C7 H16 - CH4 + O
C6 H12 O C6 H13 N

Calculating M+1 and M+2

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Isotope Abundance patterns for some
atoms

Can be diagnostic
for some isotopes

Isotope
peaks

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Interpreting Mass-Spectral
Fragmentation Patterns
The way molecular ions break down can
produce characteristic fragments that help in
identification
Serves as a fingerprint for comparison with
known materials in analysis (used in forensics)
Positive charge goes to fragments that best can
stabilize it

Fragmentation process
There are 3 type of fragmentations:
1) Cleavage of s bond
+.
---- C C ---- ---- C + +
. C ----

At heteroatom
+.
---- C Z ---- ---- C + +
. Z ----

a to heteroatom
+.
---- C - C Z ---- C=Z + + ---- C .

+.
---- C - C Z ---- Z+. + ---- C = C

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Fragmentation process
2) Cleavage of 2 s bond (rearrangements)

+.
---- HC C Z ---- ---- C=C + + HZ

Retro Diels-alder CH2


+. +. CH2
+ CH2
CH2

McLafferty
H H
Z +. Z +. CH2

CH2
Z R Z R

Fragmentation process

3) Cleavage of Complex rearrangements

Fragmentation rules in MS

1. Intensity of M.+ is Larger for linear chain


than for branched compound

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2. Intensity of M.+ decrease with
Increasing M.W. (fatty acid is an
exception)
3. Cleavage is favored at branching
reflecting the Increased stability of
the ion
Stability order: CH3+ < R-CH2+ < R2-CH+
< R3-C+
R
R CH
R

Loss of Largest Subst. Is most favored

Illustration of first 3 rules (large MW)

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Branched alkanes
H3C H3C H3C
CH3
H3C
CH3
H3C

MW=170

M.+ is absent with heavy branching


Fragmentation occur at branching: largest fragment loss

Illustration of first 3 rules


(Linear alkane with Smaller MW)
Molecular ion is stronger than
in previous sample

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Illustration of first 3 rules
(Branched alkane with Smaller MW)
43 Molecular ion smaller than
linear alkane

Cleavage at branching is
favored

Hexane

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Mass Spectral Fragmentation of
Hexane
Hexane (m/z = 86 for parent) has peaks at m/z =
71, 57, 43, 29

Rule 3 Alkanes

Cleavage Favored at branching

Loss of Largest substituent


Favored

Rule 1: intensity of M.+


is smaller with branching

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Fragmentation in different isomers

Molecular ion almost absent

Practice Problem: methylcyclohexane


or ethylcyclopentane?

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Fragmentation rules in MS
4. Aromatic Rings, Double bond,
Cyclic structures stabilize M.+
5. Double bond favor Allylic Cleavage
Resonance Stabilized Cation

-R
.
+ +
H2C CH CH2 R H2C CH CH2

H2C CH CH+2

Aromatic ring has stable M.+

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Cycloalkane
ring has
stable M.+

Fragmentation rules in MS
6. a) Saturated Rings lose a Alkyl Chain
(case of branching)
R
+. +
-R.

b) Unsaturated Rings Retro-Diels-Alder


CH2
+ . +. CH2
+ CH2
CH2

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Retro Diels-alder
+. +.

Fragmentation rules in MS
7. Aromatic Compounds Cleave in b
Resonance Stabilized Tropylium
+
CH2 CH2
R
C
-R. CH
+
+
CH

Tropylium ion +

m/z 91

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Tropylium ion
+

Fragmentation rules in MS
8. C-C Next to Heteroatom cleave leaving the
charge on the Heteroatom
+
x - [RCH2] H2C Y R
R CH2 CH2 Y R

+
H2C Y R

R2 R1 R1 R1
- [R2 ] C C
+
C
larger
+ O
O O
x

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Fragmentation rules in MS

9. Cleavage of small neutral molecules


(CO2, CO, olefins, H2O .)
Result often from rearrangement
McLafferty
H
H H O

x
x
- CH2=CH2
x

O CH2 O CH2
C
C CH2 C CH2 Y CH2
Y CH2 Y CH2

Ion Stabilized
Y H, R, OH, NR2
by resonance

Alkenes
Most intense peaks are often:
m/z 41, 55, 83

Rule 4: Double Bond Stabilize M+

Rule 5: Double Bond favor


Allylic cleavage

-Et
CH2 + CH CH Et +CH
2 CH CH CH2 CH CH +
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Me Et Me Et Me Et
M+ = 112 m/z = 83

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Alkenes

Aromatic
compound

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Mass Spectral Cleavage
Reactions of Alcohols
Alcohols undergo a-cleavage (at the bond next
to the C-OH) as well as loss of H-OH to give
C=C

Hydroxy compounds
R1
R1 R1 +
- R3
x
+ C O H
R2 C O H C O H
R2 R2
R3 If R1=H m/z 45, 59, 73

Loss of largest group If R1=alkyl m/z 59, 73, 87

H H CHR
+ CHR
CHR OH
x

CHR OH (H2O)
+ +
CHR CHR CHR CHR
CHR CHR
CHR CHR CHR CHR
CHR CHR M (H2O)

H
CHR
x

OH - H2O CHR
- CHR=CHR
CHR CHR CHR
CHR M (H2O) (C1=C2) Alkene

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Alcohols

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Phenol

Aromatic Ether Molecular ion is prominent

+
O x
O
R -CO
1) - R C5H5

m/z 93 m/z 65
Cleavage in b of aromatic ring
O O

2) - CH2=CH2 x

H
H
H
Rearrangement m/z 94
O
H
x

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B
Aliphatic Ether
Cleavage of C-C next ot Oxygen
Loss of biggest fragment

+
CH3 CH2 OCH2 CH2 CH2 CH3 CH3 CH2 O+ =CH2

m/z 59 CH3 CH2 O CH2+

m/z 45
Ether
Rearrangement
B
m/z 73

1- Cleavage of C-C next to Oxygen H CH2

CH3 CH2CH O CH2 CH3 CH =O+ CH2 Box rearr.


CH3 CH3
M+ m/z 73 CH =O+ H

2- Cleavage of C-O bond: charge on alkyl CH3


m/z 45

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Fragmentation of Ketones and
Aldehydes
A C-H that is three atoms away leads to an
internal transfer of a proton to the C=O, called
the McLafferty rearrangement
Carbonyl compounds can also undergo a
cleavage

Mass Spectral Cleavage of


Amines
Amines undergo a-cleavage,
generating radicals

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Ionization
Techniques
Electron Impact (EI)
Under these conditions, very energetic Ions are produced
=> Fragmentation M.+ is often weak
e.g. ROH often loose H2O
.
Intensity of M + Depends on the stability of the Ion
Most Stable
Ketones
Aromatic
Amines
Conjugated Alkenes
Ester
Cyclic compounds
Ether
Sulfides
Acid ~ Amides ~ Aldehydes
Short Linear Alkanes
Mercaptans
Least Stable
M.+ is often absent in: { Alcohols,
Nitriles, Nitrates, Nitro

Other Mass Spectral Features


If parent ion not present due to electron
bombardment causing breakdown, softer
methods such as chemical ionization are
used
Peaks above the molecular weight appear
as a result of naturally occurring heavier
isotopes in the sample
(M+1) from 13C that is randomly present

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Other Ionization Techniques
Under EI (Electron Impact), the molecular ion may be weak or absent
In such case, the best solution is to run CI Chemical Ionization
Which result in intense M+1 ion with little fragmentation
CI Chemical Ionization
Vaporized Sample is introduced with excess gas (usually methane)
The gas is ionized producing Primary ions that react with excess gas
to produce secondary ions
CH .+ + CH CH + + CH .
4 4 5 3
CH3+ + CH4 C2H5+ + H2
Secondary ions react with sample M:
CH5+ + M [M+H]+ + CH4
C2H5+ + M [M+H]+ + C2H4
Less energy than CI => less fragment

Comparing
different
techniques

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Classification according to method of separating charged
particule
1. Magnetic Field Deflection
1. Magnetic Field only: Mass 12-500 in seconds
2. Double Focussing: High resolution to 4 decimals
r = 60,000
2. Quadrupole Mass Spectrometer
1. Quadrupole Mass Filter: Mass Scanning by varying RF & DC
Frequencies
2. Quadrupole Ion Storage (Ion Trap)
- Compact
-Inexpensive
-Very Sensitive
-GC/MS

Classification according to method of separating charged


particule

3. Time of Flight
- Need Fast Electronic (10-7 s)
- Used With Sophisticated Ionization Methods
(FAB, Laser Desorption .)
4. FT-ICR (Fourier Transform Ion Cyclotron
Resonance)
- Very High Precision most expensive
5. MS/MS (Tandem Mass Spectro.)
-Specific Ions are Separated in First MS
- Pass one at a time in a collision chamber
- Second MS produce Daughter Ions

-used for large molecule and resolution of mixture

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Comparing Mass Spectrum of a compound in different
type of mass spectrometers
Magnetic sector Quadrupole

Time of Flight

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MW = 164

Sample introduction/ionization method:

Ionization Typical Sample Mass Method


method Analytes Introduction Range Highlights

Hard method
Relatively GC or to
Electron Impact versatile
small liquid/solid 1,000
(EI) provides
volatile probe Daltons
structure info
Relatively GC or to Soft method
Chemical
small liquid/solid 1,000 molecular ion
Ionization (CI)
volatile probe Daltons peak [M+H]+
Liquid Soft method
Peptides to
Electrospray Chromatograp ions often
Proteins 200,000
(ESI) hy multiply
nonvolatile Daltons
or syringe charged
Carbohydrates Soft method
Fast Atom Sample mixed to
Organometallics but harder
Bombardment in viscous 6,000
Peptides than ESI or
(FAB) matrix Daltons
nonvolatile MALDI
Matrix Assisted
Peptides Sample mixed to Soft method
Laser
Proteins in solid 500,000 very high
Desorption
Nucleotides matrix Daltons mass
(MALDI)

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Linked Systems:

Gas chromatography coupled to mass


GC/MS: spectrometry
Liquid chromatography coupled to
LC/MS:
electrospray ionization mass spectrometry

Steps in the Identification of Unknown


Identify Molecular ion M.+
Determine Molecular Formula (odd / even mass
Compare with proton NMR ( # protons)
Identify base peak (note if even / odd)
One or two bond fragmentation
Analyze heteroatom (M+1 and M+2 )
S, Si, Cl, Br, .
Test your conclusions: in lab make derivatives (TMS
or Na or K complexes mass shift)

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Mass Spectroscopy C5H12O
MW = 88.15

Mass Spectroscopy C7H7Br


MW = 171.04

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Mass Spectroscopy C9H10 O
MW = 134.18

Mass Spectroscopy C11H12 O3


MW = 192.21

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Tentukan struktur senyawa dari data berikut :

Tentukan struktur senyawa dari data berikut :

13C quartet: singlet: triplet: doublet:


NMR: 30.6 ppm 45.2 ppm 108.5 ppm 149.3 ppm

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