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