Large group of analytical methods based on atomic

and molecular spectroscopy Spectroscopy?

The science that deals with interactions of various types

of radiation with matter. (electromagnetic radiation & matter)

Electromagnetic radiation? Wave particle duality Electromagnetic spectrum

Electromagnetic Spectrum
Encompasses an enormous range of wavelength and

frequency (energy)

Energy states of chemical species

Atoms, ions & molecules can exist only in certain discrete states, characterized by definite amounts of energy. When species change states, it absorbs/emits an amount of E exactly equal to the E difference between the states. 2. When atoms, ions & molecules absorb/emit radiation , the frequency, , or the wavelength , of the radiation is related to the energy difference between the states by: E1 - E2 = h = hc/
1.

How does it work?

How does it work?

A beam of UV and VIS light source is separated into its

components by a prism/diffraction grating. The monochromatic beam is then split into two beams of equal intensity by a half mirror. One passes through cuvette containing sample solution and the other through another cuvette containing only solvent Intensities of two beams measured and compared difference indicates absorption and presence of functional group
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BEERS LAW-ABSORPTION LAW

Absorption is proportional to the number of absorbing molecules.

Incident radiation

I0
Pathlength

Transmitted radiation

concentration

The amount of UV light absorbed is expressed as the samples molar absorptivity

(), defined by the equation:

= A/bc Molar absorptivity is a physical constant, characteristic of the paritcular substance being observed and thus characteristic of the particular p electron system in the molecule.

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

log I0/I = 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 cm1mol 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 cuvette quartz of 1 cm (Styrene, MW = 104 g/mol)
max=250 nm

How does it work?

When UV and VIS light is absorbed by the sample

chromophores (system containing the electrons responsible for absorption in UV-VIS) electrons held in bonds are readily excited to higher energy. Energy absorbed is related to the type of functionality within compound

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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

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, s 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 p orbitals, in = or bonds. These are the most readily excited and are responsible for a majority of electronic spectra in the UV-VIS regions. * p 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 excitedstate energy levels (either * s or *p ). Therefore, absorption of radiation results in an electronic transition to antibonding orbitals. The most common transition:

p n n

p* p* s*

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)

p* and n

p*

* The probability ofp Molar absorptivities,

* p * is higher than n

p*

p
n

p * (1000-100,000) p * (less than 1000)

Measurement of the spectrum

Taken in dilute solution Compound diluted in solvent of choice Solvent: transparent & less interaction with absorbing species Most common 95% ethanol Others: cyclohexane, acetonitrile
Spectrum plotted as log10 (Io/I) vs

For publication and comparison converted to vs or

log vs

Intensity measured as area under curve max maximum intensity adopted in everyday use

Selection rule and intensity

Irradiation does not always give rise to excitation of

electrons from any filled orbital to any unfilled orbital Due to symmetry governing transitions which are allowed. Intensity is a function of allowedness (electronic transition of target area to capture light)
= 0.87 x 1020 P.a P: transition probability a: target area of chromophore

The longer the conjugated system the more intense the

absorption

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 p electrons in p ) conjugation. the chromophores (n

Auxochrome
Functional group that does not itself absorb in the UV

region but has the effect of shifting chromophore peaks to longer wavelengths and increasing intensities
Ex: -OH (270nm, = 1450), NH2 (280nm, = 1430) on benzene(256nm, = 200)

Spectra changes can be classed as:

Bathochromic shift (red shift) m ax shifted to longer

wavelength. It may caused by addition of conjugation. Hypsochromic shift (Blue shift) -m axshifted 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 p bond system of the benzene ring is removed on protonation Neutral aniline absorbs at m ax = 230 nm = 8600 But in acid solution (protonated) m ax = 203 nm = 7500

Solvent effect
-*:
transition result in an excited state more polar than ground

state. Dipole interaction with solvent molecule will lower energy of excited state longer wavelength maxima Red shift from hexane to ethanol n-*: Weak transition of O lone pair in ketones (n-* transition) shows solvent effect in opposite direction Solvent effect is lesser extent to which solvents can hydrogen bond to carbonyl group in excited state blue shift

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Spectra changes can be classed as: continue.

Hyperchromic effect increased absorption

intensity ( ) Hypochromic effect decreased absorption intensity ( )

Note: m ax = the wavelength of the absorption maximum 1% 1cm = Absorption [log10 ( I 0 I )] of a 1% solution in a cell with 1 cm pathlength- used when MW of a compound is not known

Structure Determination in Conjugated Systems: UV Spectroscopy

Conjugated compounds can absorb light in the ultraviolet

region of the spectrum The electrons in the highest occupied molecular orbital (HOMO) undergo a transition to the lowest unoccupied molecular orbital (LUMO) The region from 2 x 10-7m to 4 x 10-7m (200 to 400 nm) is most useful in organic chemistry A plot of absorbance (log of the ratio of the intensity of light in over light transmitted) against wavelength in this region is an ultraviolet spectrum
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Ultraviolet Spectrum of 1,3-Butadiene

Example: 1,4-butadiene has four p molecular orbitals

with the lowest two occupied Electronic transition is from HOMO to LUMO at 217 nm (peak is broad because of combination with stretching, bending)

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Interpreting UV Spectra: The Effect of Conjugation

max: wavelength where UV absorbance for a compound is

greatest Energy difference between HOMO and LUMO decreases as the extent of conjugation increases max increases as conjugation increases (lower energy)
1,3-butadiene: 217 nm, 1,3,5-hexatriene: 258 nm

Substituents on p system increase max

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Conjugation, Color and the Chemistry of Vision

Visible region is about 400 to 800 nm Extended systems of conjugation absorb in visible

region b-Carotene, 11 double bonds in conjugation, max = 455 nm Visual pigments are responsible for absorbing light in eye and triggering nerves to send signal to brain

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