ANALISIS

FISIKO KIMIA I
Prof. Dr. Sudibyo Martono, M.S., Apt.
Spektra Massa
 Mass Spectrometry

The basic principles

Applications

SPECTRA MS 3
 Mass Spectrometry
THE MAIN USE OF MS IN ORGANIC CHEMISTRY IS:

• DETERMINE THE MOLECULAR MASS OF ORGANIC

COMPOUNDS

• DETERMINE THE MOLECULAR FORMULA OF ORGANIC

COMPOUNDS

SPECTRA MS 4
 Mass Spectrometry
• Molecular weight can be obtained from a very small
sample.

• It does not involve the absorption or emission of

light.

• A beam of high-energy electrons breaks the

molecule apart.

• The masses of the fragments and their relative

abundance reveal information about the structure of
the molecule.
SPECTRA MS 5
 What is a mass spectrometer ?

A mass spectrometer is an instrument which

produces charged particles (ions) from
chemical substances under analysis.

It then uses magnetic and/or electric fields to

separate those ions and to measure their
mass.

SPECTRA MS 6
SPECTRA MS 7
SPECTRA MS 8
SPECTRA MS 9
SPECTRA MS 10
 Ion Generation

~70 Volts
Electron Collector (Trap)

Neutral
Molecules Positive Ions
+ Inlet _
Repeller _
+ + ++ + + + To Analyzer

Electrons
e - e- e-
_

Filament Extraction
Plate

SPECTRA MS 11
SPECTRA MS 12
 APCI

SPECTRA MS 13
 Electrospray Ionization
• Sample dissolved in polar, volatile buffer (no
salts) and pumped through a stainless steel
capillary (70 - 150 m) at a rate of 10-100
L/min

• Strong voltage (3-4 kV) applied at tip along

with flow of nebulizing gas causes the
sample to “nebulize” or aerosolize

• Aerosol is directed through regions of higher

vacuum until droplets evaporate to near
atomic size (still carrying charges)
SPECTRA MS 14
SPECTRA MS 15
 ESI

SPECTRA MS 16
 Electrospray (Detail)

SPECTRA MS 17
 Electrospray Ionization
• Can be modified to “nanospray” system with
flow < 1 L/min
• Very sensitive technique, requires less than
a picomole of material
• Strongly affected by salts & detergents

• Positive ion mode measures (M + H)+ (add

formic acid to solvent)
• Negative ion mode measures (M - H)- (add
ammonia to solvent)
18
SPECTRA MS
 ELECTRON IMPACT
H H
H-C:H + e H-C H + 2e CH4
H H
RADICAL CATION
H
H H
H-C
H H-C + + H
H
BOND-BREAKING RADICAL
CATION
H
H-C + H+
H
ONLY CATIONS ARE CARRIED TO DETECTOR
SPECTRA MS 19
 Electron Impact Ionization
A high-energy electron can dislodge an electron
from a bond, creating a radical cation (a positive ion
with an unpaired e-).
H H
H C C H
H H
H H H H
e- + H C C H H C C+ H
H H H H
H H
H C+ C H
SPECTRA MS =>
20
H H
 SEPARATION OF IONS
• Only the cations are deflected by the magnetic
field.

• Amount of deflection depends on m/z.

• The detector signal is proportional to the

number of ions hitting it.

• By varying the magnetic field, ions of all masses

are collected and counted. =>

SPECTRA MS 21
 Positive or Negative Ion Mode ?
• If the sample has functional groups that readily
accept H+ (such as amide and amino groups
found in peptides and proteins) then positive ion
detection is used-PROTEINS

• If a sample has functional groups that readily lose

a proton (such as carboxylic acids and hydroxyls
as found in nucleic acids and sugars) then
negative ion detection is used-DNA

SPECTRA MS 22
 Mass Spectrometer Schematic

Sample Data
Introduction Output

Inlet Data
System

Ion Mass Ion

Source Analyzer Detector

Vacuum
SPECTRA MS Pumps
23
 The magnetic field exerts a force on these fast-
moving ions and causes them to move in a circular
path, the radius of which is dependent upon their
mass to charge ratio (m/z) and speed. 

SPECTRA MS 24
 Magnetic Mass Separation

Correct m/z ratio

ion not detected ion detected
m/z too small

Ion Detector
Source N
ion not detected
m/z too large
Electromagnet

SPECTRA MS 25
 Fragments appear due to bond cleavage next to C=O
(alkoxy group loss, -OR) and hydrogen rearrangements.
43 -OCH2CH3
100
relative abundance
80
60 -C2H3
40
20 45 61 88
0
0 20 40 60 80 100
mass / charge (m/z)

61
H
O H
m/z(parent ion) = 88
H3C O CH2CH3
SPECTRA MS
43 45 26
Alarm Pheromone of Honey Bee

Typical Mass Spectrum

100%
43
base peak given 100%
abundance
Note: 114-71 = 43
58
Note: 43 = mass of radical
50%
71
114
molecular ion

m/e
Isotope peaks - P+1,
SPECTRA MS P+2, etc 27
 Fragmentation of 2-Methylpentane

CH3

CH3CH2CH2 •
+ CHCH
+
3
Facile
m/z 43
+
CH3 • CH3
Facile
CH3CH2CH2CHCH3 CH3 •
+ CH3CH2CH2CH +

More difficult m/z 71

m/z 86
CH3
CH3CH2 • + CH
+
2CHCH3
m/z 57
SPECTRA MS 28
 Mass Spectrum of 2-Methylpentane

43
100
CH3
80
CH3CH2CH2CHCH3
Intensity

60

40
71
20
57
86, M+

10 20 30 40 50 60 70 80 90
SPECTRA MS m/z 29
 Mass Spectrum of Pentane

43
100 CH3CH2CH2CH2CH3
80
Intensity

60 42
57
40 41

20 27
29 39 72, M+

10 20 30 40 50 60 70
SPECTRA MS 30
m/z
 2-Chloroethylphenyl ether

94
100
Cl H
107
80
Intensity

O
60 94

40 156
107
20

20 40 60 80 100 120 140 160

SPECTRA MS 31
m/z
 Triple Quadrupole Mass Analyzer

Sample Inlet

Q2
Ion Guide (Collision cell)
EM
Q1 Detector
Q3

SPECTRA MS 32
 LC-MS-MS

SPECTRA MS 33
 MRM

SPECTRA MS 34
 Multiple Reaction Monitoring in Triple Quadrupole

Q1 (227) Q2 collision cell Q3 (181)

OH
NO2 OH

LC NO2
column

NH3+
NH2+

O OH

Set on mass of Fragment Transmit only

parent ion parent ion diagnostic
product ion
highest duty cycle triple quadrupole scan type!
SPECTRA MS 35
 Applications
Mass spectrometers are used for all kinds of
chemical analyses:

- Chemical analysis (Chemical Research

- Environmental analysis
- Analysis of petroleum products
- Trace metals
- Biological materials

SPECTRA MS 36
 Terima Kasih

E-mail: sudib_kekes@yahoo.co.id

SPECTRA MS 37
SPECTRA MS 38