1. Absorption Spectrometry
Particularly UV-Visible

2. One difference between certain compounds is their colour.
Quinone is yellow;
Chlorophyll is green;
2,4-dinitrophenylhydrazone derivatives of aldehydes and ketones range in colour from bright yellow to deep red, depending on double bond conjugation;
Aspirin is colourless.

3. The human eye is functioning as a spectrometer analyzing the light reflected from the surface of a solid or passing through a liquid.
We see sunlight (or white light) as uniform or homogeneous in color, it is actually composed of a broad range of radiation wavelengths in the ultraviolet (UV),
visible and infrared (IR) portions
of the spectrum.

5. Violet: 400 - 420 nm Indigo: 420 - 440 nm
Blue:   nm Green:   nm Yellow:   nm
Orange:   nm
Red:   nm
                                                                                                

6. Absorption of nm light renders a substance yellow, and absorption of nm light makes it red.
Green is unique in that it can be created by absorption close to 400 nm as well as absorption near 800 nm.

8. Wavelength is the distance between adjacent peaks (or troughs), in meters, centimeters or nanometers (10-9 meters).
Frequency is the number of wave cycles that travel past a fixed point per unit of time, and is usually in cycles per second, or hertz (Hz).
Visible wavelengths range from ~ 400 to 800 nm. The longest visible wavelength is red and the shortest is violet. Other common colors of the spectrum, in order of decreasing wavelength, may be remembered by the mnemonic: ROY G BIV.

9.  = c/  = frequency,  = wavelength,
The energy associated with a given segment of the spectrum is proportional to its frequency. The energy carried by a photon of a given wavelength of radiation:
                                                                                                                                                                                                                                                   
 = h
h = Planck’s constant = 6.6x10-34 J.sec
 = c/  = frequency,  = wavelength,
c = speed of light = 3 x 108 m/sec
Frequency remains constant
Wavelength and the speed of light change with the medium

10. Refractive index = c/v medium n* air 1.0003 Water 1.333
50% sucrose in water
carbon disulfide
crystalline quartz (no)
1.553 (ne)
diamond

11. Polarized electromagnetic wave

12. Carvone

13. Each photon of light has a distinct energy –
Causes transitions between quantized energy states in atoms, molecules etc.
Absorption
Emission
Scattering

14. Absorption Only if energy states differ by h
Other frequencies pass through
Measure decrease in P at each frequency

15. Emission Chemical species can be excited by Thermal Chemical
Electrical energy.
If the subsequent relaxation to the ground state results in the release of light –
This is emission

16. Luminescence When energy is absorbed the chemical species are excited.
The excited species will have a limited lifetime
They will relax – lose the excess energy- and return to the ground state.
If the excitation is by light and light is emitted upon relaxation – you have luminescence – fluorescence or phosphorescence.
Incoming beam is unidirectional, luminescence is emitted in all directions

17. Why is the sky blue?

18. Light passes more molecules when coming from
the horizon – so some is scattered away and sky is very pale blue

19. WHY IS THE SUNSET RED?
As the sun sets, light must travel farther through the atmosphere before it gets to you.
More of the light is reflected and scattered.
As less reaches you directly, the sun appears less bright. The color of the sun appears to change, first to orange and then to red. This is because even more of the short wavelength blues and greens are now scattered. Only the longer wavelengths are left in the direct beam that reaches your eyes.

21. Rayleigh Scattering

22. Mie Scattering For particle sizes larger than a wavelength, Mie
scattering predominates. Mie scattering is not strongly wavelength dependent and produces the almost white glare around the sun when a lot of particulate material is present in the air.
It also gives us the white light from mist and fog.

23. Tyndall effect
When a very dilute dispersion of small particles or droplets is viewed directly against an illuminating light source it may appear to be transparent.
In contrast, when the same dispersion is viewed from the side (at a right angle to the illuminating beam), and against a dark background, the dispersion may appear turbid and blue-white.
The scattered light is due to Tyndall scattering and the optical effect is referred to as the Tyndall effect.

24. Raman Scattering
Like Rayleigh scattering, Raman scattering depends upon the polarizability of molecules.
The incident photon can excite vibrational modes of the molecules, yielding scattered photons which are diminished in energy by the amount of the vibrational transition energies.
Thus the scattered light is at lower energy than the incoming light.
Occurs with particles much smaller than wavelength of light

25. An application of Raman
The scattering produced by a laser beam directed on the plume from an industrial smokestack can be used to monitor the effluent for molecules which will produce recognizable Raman lines.
We will see some Raman scattering as an interference in our fluorescence spectra.

26. Turbidity is a critical water quality parameter
many applications, from drinking water to ultrapure processes.

27. Turbidimetry Turbidity is a critical water quality parameter
Standards – formazin
(insoluble polymer)
Turbidimeter- measures
the amount of radiation
that passes forward
Nephelometer – measures scattered radiation
(good if low turbidity)
Some instruments use the ratio of these two measurements
Turbidity is a critical water quality parameter
in many applications, from drinking water
to ultrapure processes.

28. Electronic transitions- generally from HOMO to LUMO

29. Unsaturated functional groups that absorb UV or visible light are called chromophores
Single C-C bonds hold their electrons too tightly for transitions to occur.
But not so C-S, C-Br, C-I
The energy absorbed from the UV is comparable to some bond energies – some bonds can be broken – called photolysis

30. was used to color the robes of the royal and wealthy
widely distributed in plants, but is not sufficiently stable to be used as permanent pigment, other than for food coloring
A common feature of all these colored compounds, displayed below, is a system of extensively conjugated pi-electrons.

31. To Longer Wavelength
Bathochromic
To Shorter Wavelength
Hypsochromic
To Greater Absorbance
Hyperchromic
To Lower Absorbance
Hypochromic

32. Absorption Analysis
Usually done on liquids, but gases can also be analyzed

33. 1,2,4,5-tetrazine

34. Why are the bands so broad?
Within each electronic state there are numerous vibrational states. At room temperature, Molecule is in lowest vibrational state.
But it can excite into a variety of vibrational levels
In liquids additional broadening occurs because of collisions with the solvent which further reduce the lifetime of the excited state. (Short lifetime, broad peak – Heizenberg uncertainty principle)

36. Advantages of UV-visible absorption
Moderately low LOD
10-7 to 10-6 M
Two types of applications:
Trace
Micro
Compared to gravimetric-rapid and convenient
Low cost
Can automate (increases instrument cost)

37. Absorption Only if energy states differ by h
Other frequencies pass through
Measure decrease in P at each frequency

38. Beer-Lambert law
Extent of absorption depends on number of encounters between photons and absorbing species.
P0 = power of incident radiation
b = pathlength of cell

40. Percent Transmittance
A = log P0/P = -log T
A = log 1/T
%T = 100 T
Ranges from 0 to 100 %
A = log 100/%T

41. Deviations from Beer’s law
Instrumental
Non-monochromatic light
negative deviation at high conc.
Wide slits give lower A values
Stray light
a) Reflections
b) Higher orders
c) slit diffraction
All cause negative deviation

42. Real Deviations
Refractive index increases with concentration – at high concentrations there is a negative deviation
At concentrations > 0.01 M, each molecule affects the charge distribution of its neighbour
This can alter the ability of the molecules to absorb light.
Can also occur with high concs of surrounding electrolyte

43. Chemical Deviations
Equilibria - acid base pH control
Activity coef.
Temperature
Solvent effects

44. Equilibrium

45. Analysis of two component sample
If the components have absorptions that do not overlap, then measurement of each can be done independently of the other.

46. Analysis of two-component sample
Beer’s law is additive
The total absorbance will equal the sum of the absorbances
If two compounds, x and y, absorb at different enough wavelengths:
At 1, A1 = (Ax)1+ (Ay)1
= x1 [x] + y1[y] (assuming b=1cm)
At 2, A2 = (Ax)2+ (Ay)2
= x2 [x]+ y2 [y]

47. Determine the molar absorptivities () for x and y for each wavelength, 1 and 2
Then solve the simultaneous equations

48. If there is a lot of overlap between spectra
Am = xb[x] + yb[y] at any wavelength
Axs = xb[x]s
Ays = yb[y]s

49. The absorbances of the standards and mixtures are measured at a variety of wavelengths.
The data at a particular wavelength will give one point on the graph.
Thus measurements need to be done at at least 5 wavelengths