Identification of Compounds

by UV, IR, NMR AND MASS SPECTROMETRIES

Gas Chromatogram of Flavor Compounds

1 10 11 17

3 13 15 8 5 16

6 9 12 14

Identification of Compound
H3C H2 H2 H CH3 H H H H CH3

O
CH3

SPECTROMETRIC IDENTIFICATION

I. II. III. IV. V.

Introduction of Spectrometric Analyses Ultra Violet Spectrometry Infrared Spectrometry Nuclear Magnetic Resonance Spectrometry Mass Spectrometry

I. Introduction of Spectrometric Analyses

The study how the sample interacts with different wavelength in a given region of electromagnetic radiation is called spectroscopy or spectrochemical analysis. The collection of measurements signals (absorbance) as a function of electromagnetic radiation is called a spectrum.

Spectrum of Radiation

Electromagnetic Spectrum.
Wavelength, cm , 10-10 10-8 10-6 10-4 ultraviolet visible 10-2 1 microwave 102 104 106 108

infrared

1020

1018

1016

1014

1012

1010

108

106

104

102

frequency, (cycles/sec) , yellow orange violet green visible region

blue

400

500

600 700 Wavelength, nm ,

red

800

Power

ray

ray

radio

Energy Absorption
The mechanism of absorption energy is different in the Ultraviolet, Infrared, and Nuclear Magnetic Resonance regions. However, the fundamental process is the absorption of certain amount of energy. The energy required for the transition from a state of lower energy to a state of higher energy is directly related to the frequency of electromagnetic radiation that causes the transition.

Spectral Distribution of Radiant Energy

V' C V = Wave number (cm -1) = Wave length (nm) = Velocity of Radiation (constant) 3 1010 cm/sec = Frequency of Radiation (cycles/sec)
V 1

V' =
C

(The energy of photon) E = Vh (Planck's Constant 6.62 10-27 erg -sec ) E = Vh = h C = V

C C

1 x 107 erg = I joule =0.239 calorie Avogadros number = 6.02 x 10 23 mol-1

V =

Spectral Properties, Application and Interactions of Electromagnetic Radiation

Energy Electron volts, eV Wave Number V Wavelength Frequency Type Radiation Type spectroscopy Type Quantum Transition

Kcal/mol

cm-1

cm

Hz

9.4 x 107

4.9 x 106

3.3 x 1010

3 x 10-11

1021

Gamma ray X-ray Ultra violet Visible

Gamma ray emission X-ray absorption, emission UV absorption

Nuclear Electronic (inner shell)

9.4 x 103

4.9 x 102

3.3 x 106

3 x 10-7

1017

9.4 x 101

4.9 x 100

3.3 x 104

3 x 10-5

1015

Electronic (outer shell) Molecular vibration

9.4 x 10-1

4.9 x 10-2

3.3 x 102

3 x 10-3

1013

Infrared

IR absorption

Molecular rotation

9.4 x 10-3

4.9 x 10-4

3.3 x 100

3 x 10-1

1011

Microwave Radio

Microwave absorption Nuclear magnetic resonance Magnetically induced spin states

9.4 x 10-7

4.9 x 10-8

3.3 x 10-4

3 x 103

107

Molecular Orbital of Triplet Oxygen

Molecular Atomic
* 2Px 2Py 2Pz * * 2Pz 2Py 2Px

Atomic

Energy
* 2S 2S

* 1S 1S

Quantum Numbers and Orbitals

Principal quantum number (n): The average distance of the electron from the nucleus. 1, 2, and so on. Azimuthal quantum number (Momentum quantum number) (l): The shape of the orbital. 1s, 2s, 2p, 3s, 3p, 3d, etc Magnetic quantum number (Orientational quantum number) (ml): The orientation of orbital in the space. 2px 2p , 2pz x, y and z are orientational quantum number
y

Electron spin quantum number (ms)= The two possible orientation of the electron in a magnetic field. 1/2 or 1/2

One S and Three P Orbitals

The 5 d Orbitals

II. Ultra Violet Spectrometry

The absorption of ultraviolet radiation by molecules is dependent upon the electronic structure of the molecule. So the ultraviolet spectrum is called electronic spectrum.

Spectral Properties, Application and Interactions of Electromagnetic Radiation

Energy Electron volts, eV Wave Number V Wavelength Frequency Type Radiation Type spectroscopy Type Quantum Transition

Kcal/mol

cm-1

cm

Hz

9.4 x 107

4.9 x 106

3.3 x 1010

3 x 10-11

1021

Gamma ray X-ray Ultra violet Visible

Gamma ray emission X-ray absorption, emission UV absorption

Nuclear Electronic (inner shell)

9.4 x 103

4.9 x 102

3.3 x 106

3 x 10-7

1017

9.4 x 101

4.9 x 100

3.3 x 104

3 x 10-5

1015

Electronic (outer shell) Molecular vibration

9.4 x 10-1

4.9 x 10-2

3.3 x 102

3 x 10-3

1013

Infrared

IR absorption

Molecular rotation

9.4 x 10-3

4.9 x 10-4

3.3 x 100

3 x 10-1

1011

Microwave Radio

Microwave absorption Nuclear magnetic resonance Magnetically induced spin states

9.4 x 10-7

4.9 x 10-8

3.3 x 10-4

3 x 103

107

Electronic Excitation

The absorption of light energy by organic compounds in the visible and ultraviolet region involves the promotion of electrons in , , and n-orbitals from the ground state to higher energy states . This is also called Energy Transition. These higher energy states are molecular orbitals called antibonding.

Types of Bonds
* Antibonding Antibonding Nonbonding Bonding Bonding

Energy

n *

n *

Electronic Molecular Energy Levels

The higher energy transitions ( *) occur a shorter wavelength and the low energy transitions ( *, n *) occur at longer wavelength.

Electronic Ground and Excitation States

hv

Energy
* hv

hv

* hv n

* n

Chromophore
Chromophore is a functional group which absorbs a characteristic ultraviolet or visible region. 210 nm 233 nm 268 nm 315 nm Double Bonds Conjugated Diene Conjugated Triene Conjugated Tetraene

and * orbitals

and * orbitals

III. Infrared Spectrometry

Radiation energy in the infrared region is absorbed by the organic compound and converted into energy of molecular vibration. The energy absorption pattern thus obtained is commonly referred to as an infrared spectrum which has the plot of intensity of radiation absorption versus wavelength of absorption.

Spectral Properties, Application and Interactions of Electromagnetic Radiation

Energy Electron volts, eV Wave Number V Wavelength Frequency Type Radiation Type spectroscopy Type Quantum Transition

Kcal/mol

cm-1

cm

Hz

9.4 x 107

4.9 x 106

3.3 x 1010

3 x 10-11

1021

Gamma ray X-ray Ultra violet Visible

Gamma ray emission X-ray absorption, emission UV absorption

Nuclear Electronic (inner shell)

9.4 x 103

4.9 x 102

3.3 x 106

3 x 10-7

1017

9.4 x 10

4.9 x 10

3.3 x 10

3 x 10

-5

10

1 5

Electronic (outer shell) Molecular vibration

9.4 x 10-1

4.9 x 10-2

3.3 x 102

3 x 10-3

1013

Infrared

IR absorption

Molecular rotation

9.4 x 10-3

4.9 x 10-4

3.3 x 100

3 x 10-1

1011

Microwave Radio

Microwave absorption Nuclear magnetic resonance

9.4 x 10-7

4.9 x 10-8

3.3 x 10-4

3 x 103

107

Magnetically induced spin states

Some Molecular Vibrations

H H O Unsymmetrical bend

Symmetrical bend

C H Stretch

Atom, Group, and Molecular Rotations

Z H atom rotation H H CH 3 group rotation H C O H OH group rotation O Y COOH group rotation

Molecular rotation

Center of gravity of the molecule is at the origin

Infrared Spectrum

Infrared Absorption and Functional Groups

3.4 6.0 10.3 5.8 3.7 2.9

m m m m m m

Alkane cis-Double Bond trans-Double Bond Carbonyl Hydroxyl Stretching of Acid Group Hydroxyl