Light = particles called photons.

Energy is inversely proportional to wavelength and directly proportional to wavenumber.

Red light, longer wavelength than blue light, is less energetic than blue light.

Molecule absorbes a photon => energy of molecule increases. Energy -> excited state.
Molecule emits a photon => energy of molecule is lowered. Energy -> ground state.

Infrared radiation stimulates vibrations.

Visible and ultraviolet radiation promotes electrons to higher-energy orbitals.
When light is absorbed by a sample, the irradiance of the beam of light is decreased.
Irradiance, P, is the energy per second per unit area of the light beam.
Light is passed through a monochromator (a prism, a grating, or even a filter) to select one
wavelength. Light of a single wavelength is said to be monochromatic, which means one
color. The monochromatic light, with irradiance P0, strikes a sample of length b. The irradiance
of the beam emerging from the other side of the sample is P.
Transmittance, T, is defined as the fraction of the original light that passes through the sample.
T = P/P0
Absorbance, A, is defined as A = log (P0/P) = -log T
When no light is absorbed, P = P0 and A = 0. If 90% of the light is absorbed, 10% is transmitted
and P = P0/10. This ratio gives A = 1.
Absorbance is so important because it is directly proportional to the concentration, c, of the lightabsorbing species in the sample.
Beer-Lambert law or Beers law: A = ebc
c: concentration of the sample (M)
b: pathlength (cm)
e: molar absorptivity/how much light is absorbed at a particular wavelength(/M*cm)
The greater the molar absorptivity,e, the greater the absorbance.
Beers law states that absorbance is proportional to the concentration of the absorbing
species. It applies to monochromatic radiation8 and it works very well for dilute solutions
(smaller or equal to 0.01 M) of most substances.
However, the absorptivity changes with concentration because properties of a molecule
will change at very high concentration (nonabsorbing solutes in a solution can begin to
interact with absorbing species and alter the absorptivity).

Moles reacted = I(A)*t(s)/(n(electrons/molecule)*F)

Often, during electronic spectroscopy, the electron is excited first from an initial low energy state
to a higher state by absorbing photon energy from the spectrophotometer.
Once in the excited state, the electron has higher potential energy and will relax back to a lower
state by emitting photon energy (this is called fluorescence)

Components of spectrophotometers include the source, sample cell, monochromator,

and detector.
Light source => Monochromator => Sample => Detector => Display
Most flame spectrometers use a premix burner, in which fuel, oxidant, and sample are
mixed before introduction into the flame. Sample solution is drawn into the pneumatic
nebulizer by the rapid flow of oxidant (usually air) past the tip of the sample capillary.
Liquid breaks into a fine mist as it leaves the capillary. The spray is directed against a
glass bead, upon which the droplets break into smaller particles. The formation of small
droplets is termed nebulization. A fine suspension of liquid (or solid) particles in a gas
is called an aerosol.

Principal differences between atomic and ordinary molecular spectroscopy lie in the
light source (or lack of a light source in atomic emission), the sample container (the
flame, furnace, or plasma), and the need to subtract background emission.

Atomic absorption spectroscopy: procedure for determination of chemical

elements employing the absorption of light (Beer-Lambert law) or determination
of the concentration of a particular element (analyte) in a sample.
o In general, each wavelength ~ only one particular element.
o Hollow cathode lamps (most common radiation source), filled with Ne or
Ar. When high V is applied b/w the anode and cathode, gas is ionized and
Cation/(+) ions strike -> toward the cathode = metal atoms from the
cathode sputtered into gas phase => collisions with high-energy electrons
emit photons (this radiation has same frequency absorbed by analyte in
flame).
o Monochromator: (purpose) to select one line from the hollow-cathode
malp and to reject as much emission from the flame or furnace as
possible.

Atomic fluorescence spectroscopy = measure the emitted light as (energy is

released as light = flurescence). Intensity of floresced light is directly proportional
to the [concentration] of atoms.

Atomic emission spectroscopy: use intensity of light emitted from flames,

plasmas, or sparks at a particular wavelength to determine the quanitty of an
elemnt in a sample.
o Intensity of emitted light is directly proportional to [concentration] of atom.
o The wavelength of the atomic spectral line => identity of the element
o A sample of a material (analyte) is brought into the flame as a gas or sprayed
solution. The heat from the flame evaporates the solvent and breaks chemical
bonds to create free atoms. The thermal energy also excites the atoms into
excited electronic states that subsequently emit light when they return to the
ground electronic state. Each element emits light at a characteristic wavelength,
which is dispersed by a grating or prism and detected in the spectrometer.
o

Sources: flames (most common), plasmas,

HPLC High Performance Liquid Chromatography

Principle: to isolate/separate the desired analyte from species that would interfere in the
analysis
Mobile phase: the solvent moving through the column; either a liquid or a gas
Stationary phase: the one that stays in place inside the column; commonly a liquid that
bonded to the inside of the tube. The function of the stationary phase in the column is to
separate different components, causing each one to exit the colum at a different time
(retention time).
In: Fluid entering the column (eluent)
Out: Fluid emerging from the end of column (eluate)
Elution: the process of passing liquid/gas through a chromatography column
HPLC uses high pressure to force solvent through closed columns => give high-resolution
separation.
HPLC system consists of:
A solvent delivery system
A sample injection valve
A high-pressure column
A detector
Display i.e. a computer
Decreasing particle size => more uniform flow through the column + less distance solute must
diffuse => increasing resolution (sharper peaks)
Small particle sizes benefit: analyte is not diluted so much as it travels through the column
The Column: expensive and easily degraded
Increasing temperature => decreases retention times, improves resolution by hastening
diffusion of solutes.
However, increased temperature => degrades stationary phase, decrease column lifetime.
Solvent more polar, greater its eluent strength (the measure of the solvent adsorption energy) =>
the more rapidly the solutes will be eluted from the column.
Normal-phase chromatography:
Polar stationary phase
More polar solvent => higher eluent strength
Reversed-phase chromatography:
Nonpolar stationary phase
Less polar solvent has higher eluent strength


Gradient elution: continuous change of solvent composition (increasing amounts of
solvent B are added to solvent A) to => increase eluent strength. Gradient elution in
HPLC is analogous to temperature programming in gas chromatography. Increased
eluent strength is required to elute more strongly retained solutes.

In gas chromatography, gaseous analyte is transported through the column by a gaseous mobile phase,
called the carrier gas.
Gas-liquid partition chromatography: stationary phase is a nonvolatile liquid bonded to the inside of the
column.
Gas-solid absorption chromatography: analyte is adsorbed directly on solid particles of stationary
phase.
Open tubular columns:
offer higher resolution
shorter analysis time,
greater sensitivity than packed columns
have less sample capacity
Wall-coated column: < 1um thick liquid coating on inside of silica tube
Support-coated: 30 um thick coating of liquid coated support on inside of silica tube

Narrow columns (higher resolution) than wider columns, but require higher operating pressure
and have less sample capacity.
Increaseing thickness of the statinoary phase => increases retention time, sample capacity and
the resolution.
Like dissolves like: nonpolar columns are best for nonpolar solutes.
Packed columns: liquid or solid stationary phase
Compared with open tubular columns, packed columns provide
greater sample capacity
But broader peaks
longer retention times
less resolution
Carrier Gas: Helium (most common and compatible with most detectors)
N2, He, H2
H2 and He give better resolution (smaller plate height) than N2 because solutes diffuse more
rapidly through H2 and He than through N2.
He is more efficient, non-flammable (H2 is flammable), works with great number of detectors.
Thusly, He is most common carrier gas used.
In temperature programming, the temperature of a column is raised during the separation to increase solute
vapor pressure and decrease retention times of late-eluting components.
Sometimes, the less volatile compounds may not even be eluted from the column. But, if the
temperature is increased at a constant rate, all compounds will be eluted and the separation of peaks is fairly
uniform.
** CAUTION: if temperature is raise too high, analyte and stationary phase will decompose.