Physical Biochemistry Raman Spectroscopy

A form of vibrational spectroscopy

An electric field will distort the molecular orbital

This is a weak effect that grows with the square of intensity
A strong electric field can induce an electric dipole moment
A varying EM field will result in the induced dipole varying
EMR has an oscillating electric field which will create an oscillating electric dipole moment, which
can scatter EMR.

Raman Scattering:

Scattered photons can be emitted in any direction, and may have

a different frequency to the excitation photon. If the scattered
photon has the same frequency as the excitation photon, then
Rayleigh scattering is occurring. It is an elastic interaction; there
is no non-kinetic transfer of energy between molecule and photon.
vsc ≈ vex

Raman Scattering is an inelastic interaction; energy is transferred between the molecule and the
photon. vsc ≠ vex.
 Stokes lines (Stokes scattering) – Energy of molecule increases: vsc < vex
 Anti-Stokes lines – Energy of photon increases: vsc > vex

Energy Level Diagrams:

At room temperature, most molecules will be is the v = 0 or 1 vibrational energy states.
In scattering, electrons are not actually promoted to a higher energy state, but we can use virtual
energy states. They do not correspond to an actual absorption.

A pictorial representation –
not the actual process.

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Physical Biochemistry Raman Spectroscopy

Raman Spectra:

Transitions between vibrational / rotational levels will lead to spectral lines on either side of the
excitation line.
The spectra on the Stokes side will be more intense than that of the anti-Stokes side. Stokes
scattering is more likely as there will be more molecules in the v = 0 state than v = 1 at room
temperature.
Vibrational spectra is centred at the wavelength of the source (Raleigh)

Raman Versus IR:

 Aqueous solutions can be used
 Any wavelength can be used
 Requires variation in polarisability (some normal modes that are not visible in IR might be
visible in Raman) e.g. O2 – has variation in polarisation, so can be used in Ramen, but
could not be used in IR. Homonuclear diatomic molecules are good examples of this.

Some normal modes are invisible to both Ramen and IR.

Both Ramen and IR data is presented as transmission (a negative curve). 100 - %Abs.
A comparison of IR and Ramen spectra is useful, as some bonds will only be visible in one or the
other. Raman is often more simple (fewer peaks) and therefore easier to interpret.

Raman Spectrometer:

High intensities are required, as this is a weak

effect; provided by laser illumination that is
focused onto the specimen.
Light from the illuminated spot on the sample
is collected using a lens, then scattered using
a monochromator. FT techniques cannot be
used.
Resonance Raman scattering can also be
used. (Needed?)

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