INSTRUMENTAL ANALYSIS

CHEM 4811

CHAPTER 9
DR. AUGUSTINE OFORI AGYEMAN
Assistant professor of chemistry
Department of natural sciences
Clayton state university
CHAPTER 9


MASS SPECTROMETRY I

PRINCIPLES AND APPLICATIONS
PRINCIPLES
Technique involves

- Creating gas phase ions from the analyte atoms or molecules


- Separating the ions according to their mass-to-charge ratio (m/z)


- Measuring the abundance of the ions
PRINCIPLES
Technique can be used for

- Qualitative and quantitative analysis


- Providing information about the mass of atoms and molecules


- Molecular structure determination (organic & inorganic)


- Identification and characterization of materials
PRINCIPLES
- Instrument is mass spectrometer

- Separates gas phase ionized atoms, molecules, and fragments
of molecules

- Separation is based on the difference in mass-to-charge ratio (m/z)

m = unified atomic mass units (u)

1 dalton (Da) = 1 u = 1.665402 x 10
-27
kg

z = charge on the ion (may be positive or negative)
PRINCIPLES
- Analyte molecule can undergo electron ionization

M + e
-
M
+
+ 2e
-

- M
+
is the ionized analyte molecule called molecular ion

- Radical cation is formed by the loss of one electron

- Computer algorithms are used to deconvolute m/z values of multiply
charged ions into the equivalent mass of singly charged ion

- Permits easy determination of molecular weight of analyte
THE MASS SPECTRUM
- A plot of relative abundance vs m/z

- The most abundant peak is known as the base peak

- The base peak is scaled to 100

- Spectrum shows fragmentation patterns

- The m/z values and the fragmentation pattern are used to determine
the molecular weight and structure of organic compounds

- Provides the accurate mass of a given isotope not the
weighted average
RESOLVING POWER
- The ability of a mass spectrometer to separate ions of two
different m/z values

- Resolving power = M/M

- M = mass of one singly charged ion

- M = difference in mass between M and the next m/z value

- The resolving power of ions in the 600 range = 600

- The resolving power of ions in the 1200 range = 1200
RESOLVING POWER
- Two methods used to calculate M

- Full width at half maximum (FWHM) = M

- 10% valley (overlap should not be > 10%)
RESOLUTION
- The value of M at a given M

- Expressed in ppm
INSTRUMENTATION
Main components of the mass spectrometer

- Sample input system

- Ionization source

- Mass analyzer

- Detector

- Vacuum pumps

- Computer based data acquisition and processing system
SAMPLE INPUT METHODS
Gas Expansion

- Useful for gas samples and liquids with sufficiently high
vapor pressures

- The gas or vapor expands into an evacuated and heated vessel

- Sample leaks through holes in a gold foil seal into the ionization
source (termed molecular leak inlet)

- Pressure in ionization is maintained at 10
-6
10
-8
torr
SAMPLE INPUT METHODS
Direct Insertion Probe

- For liquids with high boiling points and solids with sufficiently
high vapor pressure

- The probe (with the sample in a glass capillary at the tip) is
inserted into the ionization source

- The probe is electrically heated to vaporize the sample

- This method has a problem with contamination
SAMPLE INPUT METHODS
Direct Exposure Probe

- Sample is first dissolved in a solvent

- A drop of solution is placed at the rounded glass tip of the probe

- Solvent evaporates leaving a thin film of sample

- The tip is inserted into the ionization source and heated to
vaporize sample

- Less likely to be contaminated
SAMPLE INPUT METHODS
Chromatography and Electrophoresis Systems

- Chromatographic instruments are used to separate mixtures of
gases and liquids

- Separated components are introduced into a mass spectrometer
for detection

- The GC-MS system

- LC-MS system is used for nonvolatile organic compounds

- Capillary electrophoresis (CE) can also be coupled to MS
IONIZATION SOURCES
Electron Ionization (EI)

- Commonly used for analysis of organic samples

- Electrons are emitted from a heated tungsten filament cathode

- Electrons are accelerated towards the anode with a potential
of about 50 100 V

- Electrons meet at right angles with the sample molecules

- Interaction with the high energy electrons causes ionization of
sample molecules and fragmentation into smaller ions
IONIZATION SOURCES
Electron Ionization (EI)

- Referred to as hard ionization source due to the high energy
EI source

- Ions are accelerated into the mass analyzer by an accelerating
voltage of ~ 10
4
V

- Both negative and positive ions are formed by EI

- Negative ions form from molecules containing acid groups or
electronegative atoms
IONIZATION SOURCES
Electron Ionization (EI)

- Collision between ions and molecules may also result in ions
with higher m/z values than the molecular ion

An example is the (M+1) peak
- Reaction between analyte molecule and H
+
to form MH
+

or (M+H)
+
in which charge equals a+1

- Low pressure in the ionization source minimizes reaction
between ions and molecules
IONIZATION SOURCES
Chemical Ionization (CI)

- A large excess of reagent gas (1000 10000 times) is introduced
into the ionization region

- Pressures in source are typically higher than EI

- Electrons are allowed to bombard the gas-sample mixture

Examples of reagent gas
- Methane, ammonia, isobutane
IONIZATION SOURCES
Chemical Ionization (CI)

- Reagent gases are much more likely ionized by the electrons
than sample due to large excess

- Sample molecules are subsequently ionized by collision with
ionized reagent gas molecules

- Considered soft ionization source

- Less fragmentation and molecular ion is much more abundant

- Combination of CI and EI spectra provide good interpretation
IONIZATION SOURCES
Chemical Ionization (CI)

- For methane reagent gas
electrons with n interactio upon formed are CH and CH
3 4
+ + -
- + + -
+ +
3 5 4 4
CH CH CH CH
2 5 2 4 3
H H C CH CH + +
+ +
Proton transfer occurs when sample molecules collide with
+ +
5 2 5
H C and CH
IONIZATION SOURCES
Chemical Ionization (CI)
4 5
CH MH CH M + +
+ +
The following may occur if analyte is a saturated HC
4 2 5 2
H C MH H C M + +
+ +
2 4 5
H CH H) - (M CH M + + +
+ +
6 2 5 2
H C H) - (M H C M + +
+ +
29) (M m/z with ) H C (M H C M
5 2 5 2
+ = + +
+ +
IONIZATION SOURCES
Atmospheric Pressure Ionization (API) Sources

- Two major types
Electrospray Ionization (ESI)
and
Atmospheric Pressure Chemical Ionization (APCI)

- Operate at atmospheric pressure

- Modified version of ESI is the Ion Spray Source

- Used for mixtures of nonvolatile high molecular weight compounds
IONIZATION SOURCES
Atmospheric Pressure Ionization (API) Sources

Applications
- Pharmaceutical chemistry
- Biochemistry
- Clinical biomonitoring


Electrospray
- Fine spray of positively or negatively charged droplets
IONIZATION SOURCES
Desorption Ionization

- For direct ionization of solids

- Excellent tool for analysis of large molecules

- Solid samples are placed on a support and then bombarded
with ions or photons

- Different types are available
IONIZATION SOURCES
Desorption Ionization

Desorption Chemical Ionization
- Used for nonvolatile compounds

- Sample is directly introduced into the chemical ionization
source on a tungsten or rhenium wire


Secondary Ion Mass Spectrometry (SIMS)
- For surface analysis
- For large molecules
IONIZATION SOURCES
Desorption Ionization

Laser Desorption Ionization

- Uses pulsed laser

- Provides selective ionization by choosing appropriate

- Laser is focused on a solid surface to ionize material

Examples of Lasers
- IR laser: CO
2
laser
- UV laser: Nd:YAG (yttrium aluminum garnet)
IONIZATION SOURCES
Desorption Ionization

Matrix-Assisted Laser Desorption Ionization (MALDI)

- Matrix disperses large amounts of energy absorbed by the laser

- Minimizes fragmentation of the molecule

- Permits analysis of molecular weight over 10,000 Da

- Used for study of polymers, proteins, peptides
IONIZATION SOURCES
Desorption Ionization

Matrix-Assisted Laser Desorption Ionization (MALDI)

Matrix
- must be stable in vacuum and not react chemically

- must absorb strongly at laser (where analyte absorbs weakly)

Examples
- IR region : urea, alcohols, carboxylic acids
- UV region: 3-hydroxypicolinic acid, 5-chlorosalicylic acid
IONIZATION SOURCES
Desorption Ionization

Fast Atom Bombardment (FAB)

- Employs fast moving neutral inert gas atoms (Ar) to ionize
large molecules

- Sample is dissolved in glycerol and spread in a thin layer on
a metal probe

- Probe is then inserted into the mass spectrometer and a beam
of fast moving atoms probe the surface
IONIZATION SOURCES
Desorption Ionization

Fast Atom Bombardment (FAB)

- Used for analysis of surfactants and proteins (MW > 10,000)

- For large and thermally unstable molecules

- Technique works well at room temperature

- Simple and high sensitivity

- Sample can be recovered
IONIZATION SOURCES
Desorption Ionization

Fast Atom Bombardment (FAB)

- Modified technique is the continuous flow FAB (CFFAB)

- Sample introduction is through a fused silica capillary tube

- Solvent flows continuously and sample is introduced by
continuous flow injection

- For analysis of blood, urine, other body fluids, waste water
IONIZATION SOURCES
Inorganic MS Ionization Sources

Solid Samples

- Glow Discharge (GD) and Spark sources

- For sputtering and ionizing species from solid surfaces

- Primarily for atomic mass determination of elements

- GD has better S/N and able to sputter more material from sample
IONIZATION SOURCES
Inorganic MS Ionization Sources

Liquid Samples

- Inductively coupled plasma (ICP)

- Has high ionization efficiency

- Provides very simple mass spectra
MASS ANALYZERS
- Differentiates ions according to their m/z

- Different designs are available

Scanning Instruments
- Only ions of a given m/z pass through the analyzer at
a given time

- Magnetic Sector Mass Analyzer

- Quadrupole Mass Analyzer
MASS ANALYZERS
Simultaneous Transmission Instruments
- Allow transmission of all ions at the same time

- Time-of-flight (TOF)

- Ion Trap

- Ion Cyclotron Resonance Mass Analyzer

- Dispersive Magnetic Mass Analyzer

Tandem Mass Spectrometer (MS
n
)
- Two or more mass analyzers in sequence
MAGNETIC SECTOR MASS ANALYZER
- Gas phase molecules are ionized by a beam of high energy
electrons

- Electrons may be ejected from molecules (ionization) or bonds
in molecules may rapture (fragmentation)

- Ions are then accelerated in a field (sector) at a voltage V

- Sector can have any apex angle (60
o
and 90
o
are common)

- Most modern instruments combine both electric sector
and magnetic sector (double-focusing MS)
MAGNETIC SECTOR MASS ANALYZER
- The electric sector acts as an energy filter

- m/z range is 1 1400 for single-focusing and
5,000 10,000 for double-focusing instruments

- Energy of each ion = zV

- Kinetic energy depends on charge and voltage but not
on mass of ion

- Ions with small masses must travel at a higher velocity
than ions with larger masses
MAGNETIC SECTOR MASS ANALYZER
- For single positively charged ions
m = mass of ion
v = velocity of ion
z = charge of ion
V = accelerating voltage

- V changes as m varies such that mv
2
is constant
zV mv
2
1
2
=
1/2
m
2zV
v
|
.
|

\
|
=
m
1
v
MAGNETIC SECTOR MASS ANALYZER
- Ions enter a curved section of a homogeneous magnetic
field B after acceleration

- Ions move in a circle with radius r

- Attractive force on magnet = Bzv

- Centrifugal force on the ion = mv
2
/r

- The two forces are equal if the ion follows the radius of
curvature of the magnet
MAGNETIC SECTOR MASS ANALYZER
Substituting for v and rearranging gives
Bzv
r
mv
2
=
Bz
mv
r =
2V
r B
z
m
2 2
=
MAGNETIC SECTOR MASS ANALYZER
- Radius of circular path depends on m/z if V and B are kept constant

- Ions with different m/z travel in circles with different radii

- Basis of separation by m/z

- Ions with the right m/z reach the detector and others hit the
sides of the instrument and be lost

- Which m/z to reach the detector can be selected by varying V or B

- B is varied and V is kept constant in modern instruments
TIME OF FLIGHT (TOF) ANALYZER
- Makes use of a drift tube

- Pulses of ions are accelerated into the an evacuated drift tube
(free of field or external force)

- Velocity of an ion depends on m/z
(depends on mass if all ions have the same charge)

- Lighter ions move faster along the tube than heavier ions

- Ions are separated in the drift tube according to their velocities (v)
TIME OF FLIGHT (TOF) ANALYZER
- V = accelerating voltage

- If L is the length of tube (typically 1-2 m) and t is the flight
time of ion, then v = L/t

- Implies mass-to-charge ratio and flight time can be found from
- An ion mirror called a reflectron is used to increase resolution
1/2
m
2zV
v
|
.
|

\
|
=
2
2
L
2Vt
z
m
=
2zV
m
L t =
QUADRUPOLE MASS ANALYZER
- Separates ions in an electric field (the quadrupole field)

- Field is varied with time

- Oscillating radio frequency (RF) voltage and a constant DC
voltage are used to create the field

- These are applied to four precisely machined parallel metal rods

- The result is an AC potential superimposed on a DC potential

- Ion beam is directed axially between the four rods
QUADRUPOLE MASS ANALYZER
- Opposite pairs of rods are connected to opposite ends of a
DC source

- Ions follow an oscillating (corkscrew) path through the
quadrupole to the detector

- For a given ratio of DC to RF at a fixed frequency, only ions of
a given m/z value will pass through the quadrupole

- Other ions with different m/z values will collide with the
rods and be lost
QUADRUPOLE MASS ANALYZER
- The quadrupole acts as a filter so is often called the mass filter

- Sample must be ionized and in the gas phase

- m/z range is 1 1000 Da

- Has smaller range and lower resolution than magnetic sector
but faster

- Is the most common analyzer

- Rugged, inexpensive, and compact
MS MS (TANDEM MS) INSTRUMENTS
- Employs two or more stages of mass analyzers

- Example is two quadrupoles coupled in series

- First analyzer selects ion (precursor ion) and second analyzer
selects the fragments of the precursor ion

- Used to obtain more information about the structure of
fragment ions

- Fragment ions may be dissociated into lighter fragment ions or
converted into heavier ions by reaction with neutral molecule
ION TRAP
- A device in which gaseous ions are formed and/or stored
for periods of time

Two commercial types
- Quadrupole Ion Trap (QIT)

and

- Ion-Cyclotron Resonance Trap (ICR)
ION TRAP
Quadrupole Ion Trap (QIT)

- Also called Paul Ion Trap

- Uses a quadrupole field to separate ions

- A 3D field is created using a ring-shaped electrode between
two end cap electrodes

- A fixed frequency RF voltage is applied to the ring electrode

- The end cap electrodes are either grounded or under
RF or DC voltage
ION TRAP
Quadrupole Ion Trap (QIT)

- Ions are stored in trap by moving in trajectories between
electrodes

- This is done by changing signs of electrodes to repel ions as
they approach the electrodes

- Ions of a given m/z pass through an opening to the detector
when the RF of the ring electrode is changed

- m/z range is 10 1000 Da
ION TRAP
Fourier Transform Ion-Cyclotron Resonance (FTICR)

- Also called Penning Ion Trap

- Uses magnetic field to trap and store ions

- Consists of six conducting plates arranged as a cube

- Cubic cell is about 100 mm on a side and is located inside
a strong magnetic field

- Sample is ionized by an electron beam
ION TRAP
Fourier Transform Ion-Cyclotron Resonance (FTICR)

- The ions then move in circular orbits

- Path is perpendicular to the applied field

- The operating frequency is called the cyclotron frequency
DETECTORS
- Measure one m/z value at a time (single channel detectors)


- Multiple detectors are used for multiple ion detection


- High resolution magnetic sector instruments use multiple
detectors (called multicollectors)
DETECTORS
Electron Multiplier (EM)

- The most common detector in MS for ions


- Similar to PMT


- Very sensitive and has fast response
DETECTORS
Faraday Cup

- A metal or carbon cup serves to capture ions and store the charge


- Cup shape decreases loss of electrons


- Least expensive detector for ions


- Has long response time
DETECTORS
Array Detectors

- Used in TOF MS instruments

- Employs a focal plane camera (FPC) consisting of an array of
31 Faraday Cup

- Up to 15 m/z values can be measured simultaneously

- Exhibits improved precision compared with single channel
detectors