UV-VIS SPECTROSCOPY

General info: Spectrometry is based on the absorption of photons by the analyte. Spectrometric method: the sample solution absorbs electromagnetic radiation from an appropriate source and the amount absorbed is related to the concentration of the analyte in the solution.
Eg: copper solution is blue because it absorbs the yellow color from white light and transmits the remaining blue light. The higher[cu], the more yellow light is absorbed and the deeper the resulting blue color of the solution.

The electromagnetic spectrum- Review

A form of radiant energy that is propagated as a transverse wave Vibrate perpendicular to the direction of propagation - a wave motion to the radiation

Continue The relationship between wavelength and frequency: ; = wavelength in cm = frequency in s-1 or Hz c = velocity of light (3x1010 cm/s) The wavelength varies from a few to several meters, for UV & Vis nanometer for IR micrometer, commonly used as wave number, in cm-1

Continue

Electromagnetic radiation possesses a certain amount of energy called photon, related to the v or by:
E hv hc

E energy of the photon in ergs 34 6 . 62 10 Joule sec h = Planks constant,

Note: Shorter wavelength, greater energy UV from the sun burns you!!!!

Working ranges of the UV-Vis and IR spectra

UV
Vis Near IR Mid IR

200-380 nm
380-780 nm 0.78-2.5 m 2.5-15 m

The most analytically region

How does matter absorb radiation

Three basic processes by which a molecule can absorb radiation; all involve raising the molecule to a higher internal energy level, the increase in energy being equal to the energy of the absorbed radiation, hv . Three type of internal energy are quantized;

Rotational transition: the molecule rotates about various axes, the energy of rotation being a definite energy levels, so the molecule may absorb radiation and be raised to a higher rotational energy level. Vibrational transition: the atoms or groups of atoms within a molecule vibrate relative to each other, and the energy of this vibration occurs at definite quantized levels. The molecules may then absorb a discrete amount of energy and be raised to a higher vibrational energy level.

Electron transition: the electron of a molecule may be raised to higher electron energy. UV-VIS spectroscopy principal/theory

Energy level diagram illustrating energy changes associated with absorption of electromagnetic radiation

A = pure rotational changer (far IR) B = rotational + vibrational changes (near IR) C = rotational + vibrational + electronic transition (Vis + UV) Eo = electronic ground state E1 = first electronic exited state

Electronic Spectra and Molecule Structure

The electronic transition in the UV-VIS regions are due to the absorption of radiation by specific types of: Groups, bonds and functional groups within the molecule Kinds of transitions: Electrons in a molecule can be classified into 4 different types: i. Closed shell electron not involved in bonding ii. Covalent single bond electron, iii. Paired non bonding outer shell electron (n electrons) such as those on N, O, S, and halogens. These are less tightly held then electrons and can be excited by visible or UV radiation iv. Electron in orbitals, in = or bonds. These are the most readily excited and are responsible for a majority of electronic spectra in the UV-VIS regions. * and n electrons responsible for UV-VIS spectrum

Other electron remain/occupate in orbitals

A molecule also possesses unoccupied orbitals called antibonding orbitals; these correspond to excited- state energy levels (either * or * ). Therefore, absorption of radiation results in an electronic transition to antibonding orbitals. The most common transition:

n n

* or * *

above 200 nm occur at very short

HOMO: Highest Occupied Molecular Orbital LUMO: Lowest Unoccupied Molecular Orbital
Example, molecule H2

Example of transition in ketone (acetone)

* and n

* n * * is higher than The probability of * n Molar absorptivities, * (1000-100,000) * (less than 1000) n

ABSORPTION BY ISOLATED CHROMOPHORES

The absorbing groups (the electrons responsible for the absorption) in a molecules are called chromophores A molecule containing a chromophore chromogen The present of auxochrome enhance the absorption by a chromophore or shift the of absorption. Example: hydroxyl (OH), amine (-NH2), halogens (X) posses n electrons interact with electrons in the chromophores (n ) conjugation.

Spectra changes can be classed as:

Bathochromic shift (red shift) max shifted to shorter wavelength. It may caused by removal of conjugation. Hypsochromic shift (Blue shift) - max shifted to shorter wavelength. It may caused by removal of conjugation. Eg. The conjugation of the lone pair electrons on the N atom on aniline with the bond system of the benzene ring is removed on protonation Neutral aniline absorbs at max= 230 nm = 8600 But in acid solution (protonated) max = 203 nm = 7500

Spectra changes can be classed as: continue.

Hyperchromic effect increased absorption intensity ( ) Hypochromic effect decreased absorption intensity ( )

Note: max = the wavelength of the absorption maximum 1% 1cm = Absorption [ log ( I I ) ] of a 1% solution in a cell with 1 cm pathlength- used when MW of a compound is not known
10 0

BEERS LAW-ABSORPTION LAW

Fraction of radiant energy transmitted decays exponentially with pathlength.

Incident radiation Transmitted radiation

I0
Pathlength

b k

concentration

Transmittance in exponential form:

I T 10kb I0
log T log

is a constant (eq 1)

Putting in logarithmic form,

I kb I0

(Eq 2

BEERS LAW-ABSORPTION LAWcontinue Similar law holds for the dependence of T on the concentration,
T I 10k 'c I0

k ' is a new constant

(Eq 4)

(Eq 3)

log T log

I k ' c I0

Combining (Eq 1) and (Eq 3), describes the dependence of T on the pathlength and concentration.

I a is combined constant of k and k (Eq 5) 10abc I0 I log T log abc I0 1 I0 Omit the ve sign, and rearrange A log T log log abc T I T
Hence,

A abc

Conc in g/L
Pathlength in cm

Absorbance

Absorptivity

BEERS LAW-ABSORPTION LAWcontinue

Note: in absorption spectrum, the absorbance varies with wavelength in direct proportion to a (b & c are held constant). The product of the absorptivity and the molecular weight of the absorbing species is called the molar absorptivity, (cm 1mol 1L)

Thus

A bc

Example Amines, RNH2 react with picric acid to form amine picrates, which absorb strongly at 359 nm (=1.25 x 104 ). An unknown amine (0.1155 g) is dissolved in water and dilute to 100 mL. A 1 ml aliquot of this is diluted to 250 ml for measurement. If this final solution exhibit an absorbance of 0.454 at 359 nm using a 1.00 cm cell, what is the formula weight of the amine? What is a probable formula? Solution: A bc

0.454 1.25 104 cm 1mol 1L 1.00cm c c 3.63 105 mol / L

(3.65 105 mol / L)(0.250L) 100mL 1.00mL

9.08 104 mol in original flask 0.1155g 1 127 gmol 9.08 104 mol

Measurement of the UV Spectrum:

Using a dilute solution: 1 mg of compound with MW (100-400 g/mol) is dissolved in 100 mL volumetric flask. Hexane is used as a solvent. Portion of the solution is transferred to a cuvett quartz of 1 cm (Styrene, MW = 104 g/mol
max=250 nm

Choice of solvent Solvent consideration: cheap, good solvent transparent down to about 210 nm. Most commonly used 95% ethanol Solvent Effect Electronic Transition, *: atom do not move, however electrons of the atom and solvent molecules may reorganise Transition state is more polar than ground state due to dipole-dipole interaction with solvents, thus lower the energy of the excited state.

Eg: Styrene in ethanol (max= 270 nm) Styrene in hexane (max= 250 nm) about 20 nm shift (red shift, longer wavelength)

Electronic transition, n * (the weak transition of the O lone pair in ketones