Molecular

Mass Spectrometry
 Uses of Mass Spectrometer
• forms ions, usually positive, study
charge/mass ratio
• very characteristic fragmentation pattern
in charge/mass ratio
• data easier to interpret than IR and/or
NMR
• provides accurate MW of sample
• used to determine isotopic abundances
Mass Spectrum of Ethyl Benzene
 Components of Mass Spectrometer

Sample

Inlet Ion Mass

Detector
System Source Analyzer
10-5 - 10-8 torr

Signal
Vacuum Processor
System
Readout
Inlet System
Natural Abundances of Isotopes of Some
Common Reagents
Components of Mass Spectrometer

“Schematic of (a) an external sample

introduction system (note the that various
parts are not to scale) and (b) a sample
probe for inserting a sample directly into the
ion source.”
 Sample Handling
• batch inlet: 1-5 L surge tank
• gases and volatile liquids
• direct probe inlet: non-volatile liquids
• gas chromatographic inlet systems
• permeable porous material to release
carrier gas
 Ion Sources for
Molecular Mass Spectroscopy
 Electron Impact Source
• bombardment of sample with beam of
electrons
"An electron impact source."
 Fragmentation Patterns

"Some Typical Reactions in an Electron Impact Source."

 Mass Spectra
"Mass spectra for 1-decanol:
(a) 70-ev electron impact
(b)chemical ionization with isobutane as
reagent gas.”
Electron-impact mass
spectra of:
(a) methylene chloride
and
(b) 1-pentanol.
 Chemical Ionization Source
• reagent gas 103 - 104 times concentrated
as sample
• collisions with reagent gas ions causes
ionization
• less fragmentation, simpler spectra
• special modifications to deal with higher
pressures
 Spark Source
• rf spark source, 30 kv
• sample part of electrodes, produces
gaseous ionic plasma
 Field Ionization Source
• metallic anode
• cathode acts as slit
• separation: 0.5 to 2 mm
• 5 to 20 kv potential applied
• produces mainly M and M+1 peaks
Mass spectra for glutamic
acid:
(a) electron impact ionization,
(b) field ionization, and
(c.) field desorption
 Mass Analyzer
• resolution vs price and application
 Single-Focusing Analyzers
with Magnetic Deflection

"Schematic of a
magnetic sector
spectrometer."
 Magnetic Centripetal Force
Fm = Bzev
where Fm => magnetic centripetal force
B => magnetic field strength
v => velocity of particle
z => charge on particle
e => charge of electron
 Centrifugal Force
Fc = mv2/r
where Fc => balancing centrifugal force
r => radius of curvature of
magnetic sector
m =>mass of particle
 Kinetic Energy
KE = zeV = 1/2mv2
where KE => kinetic energy
V => accelerating potential
 Mass to Charge Ratio, m/z
F m = Fc
thus
Bzev = mv2/r
where v = Bzr/m

m/z = (B2r2e)/2V
 Mass Analyzer
Double-Focusing Analyzers
– higher resolution, need higher amplification
– 2 magnets or 1 magnet & 1 electrostatic field
Mattacuh-Herzog type double-
focusing mass spectrometer.
 Double Focus
Mass Spectrometer
 Time of Flight Analyzers
– non-magnetic separation
– detector - electron multiplier tube
– instantaneous display of results
Schematic of a time-of-flight mass
spectrometer.
 Quadrupole Analyzers
• 4 short parallel metal rods
• opposite rods same charge on dc source,
AC rf applied on top
Quadrupole Mass Spectrometer
 Ion Trap Analyzer
• Variable radio frequency voltage applied to
the ring electrode
• ions of appropriate m/z circulate in stable
orbit
• scan rf, heavier particles stable, lighter
particles collide with ring electrode
• ejected ions detected by transducer as an ion
current
Ion Trap Mass Spectrometer
 Measurement and Display of
Results
• photographic results in double-focus
• electron current from well protected
electrode
• galvanometers with sensitized paper
• strip chart recorder
• computer display
 Fourier Transform
Mass Spectrometer
• Usually trapped ion analyzer
• ions created by brief electron beam burst
• short rf pulse that increase linearly in
frequency with time
Computerized Mass Spectrometers

"A trapped
ion analyzer
cell.”
 Computerized
Mass Spectrometers
"Schematic diagram showing the timing
of
(a) the radio-frequency signal
(b) the transient image signal."
 Computerized
Mass Spectrometers
"Time domain (a) and (b) frequency or
mass domain spectrum for 1,1,1,2-
tetrachloroethane."
 Computerized
Mass Spectrometers
"A computer display of mass-spectral
data. The compound was isolated from a
blood serum extract by chromatography.
The spectrum showed it to be the
barbiturate, pentobarbital."
 Electron Impact Ionization
Process
M + e- -----> M+ + 2e-
where M+ => molecular ion
 Electron Impact Ionization
Process
Molecular Ions:
M+
(M+1)+
(M+2)+
 Electron Impact Ionization
Process
Molecular Ions:
M+
– results from removing an electron from a
molecule
 Electron Impact Ionization
Process
Molecular Ions:
(M+1)+
– results from one atom/molecular of C-13 or
H-2
 Electron Impact Ionization
Process
Molecular Ions:
(M+2)+
– small for most organics because it requires
two heavy atoms/molecule
• 1 C-13 and 1 H-2
• 2 C-13s
• 2 H-2s
– sizeable for chlorinated or brominated
compounds
 Electron Impact Ionization
Process
Molecular Ions:
– peaks for collision products: function of
concentration (pressure)
– stability of the molecular ion
• stabilized by  e- systems, cyclic

base peak
 Electron Impact Ionization
Process
base peak
– highest peak
– peak height against which all others are
measured for use in peak tables
 Determination of
Molecular Formula
distinguish between compounds of same
MW
C5H10O4 or C10H14
 Determination of
Molecular Formula
distinguish between compounds of same
MW
C5H10O4
13C 5 * 1.08% = 5.40%
2H 10 * 0.016% = 0.16%
17O 4 * 0.04% = 0.16%
-------
135peak/134peak 5.72%
 Determination of
Molecular Formula
distinguish between compounds of same
MW
C10H14
13C 10 * 1.08% = 10.8%
2H 14 * 0.016% = 0.22%
-------
135peak/134peak 11.0%
 Determination of
Molecular Formula
"Isotopic Abundance Percentages and
Molecular Weights for Various
Combinations of Carbon, Hydrogen,
Oxygen, and Nitrogen."
Calculate the ratios of the (M+1)+ to M+ peak
heights for the following compounds:
dinitrogen benzene and an olefin,
 Nitrogen Rule
• organic compounds with even MW, O
or even number of N
• odd MW, odd number of nitrogen atoms
 Fragmentation Patterns
"Some Typical Reactions in an Electron
Impact Source."
 Identification of Compounds
from Fragmentation Patterns
Fragmentation Patterns: Rules
• C - C bonds weaker than C - H bonds
• fragmentation most likely at a branch
• positive charge remains with fragment
with most branching
 Fragmentation Patterns
CH3
|
CH3 - CH2 -- C -- CH3
|
CH3
 Fragmentation Patterns
CH3
|
CH3 - CH2 -- C -- CH3 m/e = 71
+
 Fragmentation Patterns
+CH3 m/e = 15

CH3
|
+C - CH3 m/e = 57
|
CH3
CH3 - CH2 + m/e = 29
 Fragmentation Patterns
for alkenes: cleavage is favored at second
bond away from double bond

CH3 -- CH2 - CH = CH - CH3

----->
CH2 -- CH -- CH - CH3
+
m/e = 55
 Fragmentation Patterns
for aromatics: cleavage at beta bond from
ring, m/e = 91

 - CH2 --R ----->  - CH2 +

 Fragmentation Patterns
terminal non-carbon group: cleaves
non-carbon group

R -- X -----> R +

where R => Cl, Br, I OH, OR, SH, SR,

NH2, NHR, NR2
 Example:
2,2,4-trimethylpentane
CH3
|
CH3 - C - CH2 - CH - CH3
| |
CH3 CH3
 Example:
2,2,4-trimethylpentane
CH3
|
CH3 - C - CH2 - CH - CH3
| |
CH3 CH3

m/e % of base peak Fragment

57 100%
56 33
41 28
 Example:
2,2,4-trimethylpentane
CH3
|
CH3 - C - CH2 - CH - CH3
| |
CH3 CH3

m/e % of base peak Fragment

43 25
29 18
27 11
 Example:
2,2,4-trimethylpentane
CH3
|
CH3 - C - CH2 - CH - CH3
| |
CH3 CH3

m/e % of base peak Fragment

99 5
15 5
55 4
 Example:
2,2,4-trimethylpentane
CH3
|
CH3 - C - CH2 - CH - CH3
| |
CH3 CH3

m/e % of base peak Fragment

42 2
114 1
71 1
69 0.5
 MS/MS instrument
"Schematic of a tandem quadrupole
MS/MS instrument.”