UV-VIS SPECTROSCOPY

INSTRUMENTATION
Two configurations:
1.

Single beam

2.

Double beam

Basic components:
source of radiation
dispersive device
sample compartment
detector
electronics & readout
Configuration
note the position of the wavelength selector
(reduces sample heating)
Sources
Two sources are usually incorporated into most instruments:
1. Deuterium discharge lamp (~160 - 380 nm)
D2 (g)
-

electrical discharge

D2* (g)

2D (g) + hn

continuum of radiation obtained

quartz window has to be used in UV (glass absorbs emr < 325 nm)

2. Tungsten filament lamp (325 - 3000 nm)

-

spectral output of incandescent filament is black-body radiation

output intensity, wavelength distribution depends on temperature

two types:
standard W-filament lamp - 2900 K (approx.)
- 325 - 3000 nm
- glass bulb

quartz-halogen lamp

- halogen added to evacuated bulb

(increases life of lamp)
- high operating temperatures
- quartz bulb (glass would soften)

UV/VIS MOLECULAR SPECTROSCOPY

Wavelength Selectors
Wavelength selectors are required in optical spectroscopy in order to select the
wavelength of radiation that will be used for analysis.
Although we speak of a specific wavelength, no wavelength selection device is able to
select a single wavelength. These devices select a narrow wavelength range, referred
to as the spectral bandwidth or spectral band pass of the device.
Wavelength selector devices fall into two main categories:
1. filters:

absorption (visible region)

interference (ultra-violet, visible, infra-red regions)

2. monochromators: -

prism
grating

consist of -

entrance slit (width & sharpness of the image)

exit slit (spectral band pass)
dispersive device (dispersion of spectra)
optical components (mirrors, lenses, windows,
etc.)

in practice - the entrance and exit slits are made of

approximately equal widths since (at a given
resolving power) this allows maximum
throughput of radiation

spectral bandpass
- depends on the dispersive power of the
dispersion device and on the exit slit of the
monochromator

dispersion - two types to consider

- angular - the ratio of the change in the
angle of reflection or refraction to
the change in wavelength
dr/dl
- linear - the ratio of the change in distance
to the change in wavelength along
the focal plane
dx/dl
- often expressed as the reciprocal
linear dispersion (units = nm/mm)
dl/dx
(the smaller the RLD, the better
the dispersion)

resolution - a measure of the ability of a monochromator to

separate two peaks, and it is given by;
R = l/dl
where, l = average wavelength between the
peaks
dl = separation between the peaks
(typical values of R ~ 103 to 104)

instrument configuration
- several configurations -

Ebert
Czerny-Turner
Littrow
Rowland
Wadsworth
Eagle
Seya-Namioka

RADIATION DETECTORS (UV-VIS Spectroscopy)

Requirements: high sensitivity, good signal-to-noise ratio, uniform response over wavelength
region, fast response, linear response
Types:

photo-tube; photo-multiplier tube

silicon diode; diode array
photo-voltaic cell; photo-conductivity cell
charge-transfer device (CID, CCD)

Description:

Photo-tube:
- consists of a semi-cylindrical cathode and a wire anode sealed inside an
evacuated tube

- a photo-emissive substance, e.g., Ag and an alkali metal mixed with its metal
oxide or an alkali semiconductor material (Cs3 Sb, K2 CsSb, or Na2 KSb mixed
with a trace of Cs) is coated onto the surface of the cathode
- various surfaces respond differently to different wavelengths of radiation
- (photo-)electrons are emitted when e.m.r. hits the surface of the cathode
- if a potential of 90 V or greater is imposed across the electrodes the electrons
are collected by the anode and a current flows in the circuit. This current is
small and has to be amplified before it can be measured, recorded, or
displayed
- the current measured is directly proportional to the power of the beam of
radiation

Photo-multiplier tube:
- is similar to a photo-tube. Instead of one anode it has a series of anodes
called dynodes that serve to multiply the number of electrons that result from
one electron emitted from the cathode. It is a more sensitive detector than
the photo-tube

- the cathode surface is coated with the same type of photo-emissive material
as in the photo-tube
- the surfaces of the dynodes are coated with a material such as BeO, CsSb, or
GaP which emit several (at least 2 to 3) electrons for each high-energy
electron that strikes the surface of the dynode
- the dynodes are concave in shape and are positioned so that the electrons
that are emitted are focussed on to the next dynode
- if a potential of at least 90V (positive) is maintained between each succeeding
dynode, then an avalanche of electrons (e.g., 2n or n3 for each original
electron emitted, where n = number of dynodes) will be collected at the anode
- there is no need for large external amplification
- the current measured is proportional to the power of the beam of radiation
- photo-multiplier tubes are the most widely used detectors for spectroscopic
work in the UV-VIS region

Silicon Photo-diode:
- is a semi-conductor device that consists of a pn junction
- when such a device is placed in an electrical circuit there are two possible
arrangements:
- forward-biased operation: whereby the positive terminal of a dc source is
connected to the p region and the negative terminal to the n region:

- in this arrangement excess electrons in the n region and excess holes in

the p region move towards the pn junction where they annihilate each
other and a current flows in the circuit
- the conduction process continues since the negative terminal provides
electrons for the n region, and the positive terminal removes electrons
from the p region creating holes
- reverse-biased operation: whereby the positive terminal of a dc source is
connected to the n region and the negative terminal to the p region:

- in this arrangement excess electrons in the n region and excess holes in

the p region move away from the pn junction, the holes move towards the
negative terminal and the electrons move towards the positive terminal. A
non-conductive depletion layer is formed and no current flows in the circuit
- how, then, does a silicon diode act as a radiation detector?
- when photons of sufficient energy (UV-VIS region) strike the depletion
layer of a pn junction electrons and holes are formed causing the
depletion layer to become conducting. The resulting current in the
circuit can be measured
- the measured signal is proportional to the power of the beam of radiation
- these devices can be made sensitive to wavelengths from different
regions of the spectrum (400-1000 nm)
- sensitivity is less than that of a PMT but better than for a photo-tube
- sensitivity in the UV region (200-400 nm) is very low
Photo-diode Array:
- is a 2-dimensional array of individual silicon photo-diodes fabricated side-byside on a single silicon chip
- they are used in multi-channel instruments where the radiation intensity at
small band-widths can be measured simultaneously over a large range of
wavelengths by placing such an array along the focal plane of the
monochromator

Photovoltaic Cells:
- read page 546

Photoconductivity Cells
- read page 547

Charge-transfer device:
- is a metal-oxide semiconductor capacitor that consists of a layer of n-doped
semiconductor material that is separated from two electrodes by a thin layer
of silica that acts as an insulator

- when a negative charge is applied to the electrodes, a charge inversion region

is created under the electrodes
- positive holes may be stored in this region
- how does this device act as a radiation detector?
- when photons strike the semiconductor material, holes are formed. These
holes are collected in the charge inversion region, i.e., the device acts as a
capacitor
- the charge stored can be measured in two ways:
- voltage change occurring from movement of the charge from under one
electrode to another is measured (charge-injection device, CID)
- the charge is moved to a charge-sensing amplifier for measurement
(charge-coupled device, CCD)
- sensitivity, dynamic range, and signal-to-noise ratio approach those of the
photo-multiplier tube
- can be fabricated in a two-dimensional array that makes it useful for
multichannel detection

UV/VIS MOLECULAR SPECTROSCOPY

Bending of Calibration Curves
In practice a plot of Absorbance vs Concentration is not always linear for all
concentrations, particularly at high concentrations.
There are several reasons for this behaviour, i.e., bending of the calibration curve.
1. Theoretical
Beers Law assumes that the beam of radiation incident on the sample is
monochromatic in nature
A = -log (P/Po)
If this is not true, Beers Law requires that the logarithms of the radiant powers of
the component wavelengths be additive.
S Ali = - S log (Pi/Poi)
Since spectrophotometers allow a finite group of wavelengths (equivalent to the
band-width of the instrument) to enter the detector, it is actually the additive radiant
powers of the different wavelengths that is measured. Only when the absorption
curve is essentially flat over the spectral bandwidth employed, will Beers law hold
true. When this is not true, the calibration curve will bend towards the concentration
axis, and this becomes very noticeable at high concentrations. Additionally, the
wider the bandwidth of radiation entering the detector, the greater will be the
deviation from linearity.
S (Pi/Poi)

2. Physical Interferences
Beers law is a limiting law and applies to solutions of low concentrations only.
i) Refractive Index Effect
In the equation A = abC or ebC, it is not really a or e that is constant, but a
quantity
ah/(h2 + 2)2
where,

h = the refractive index of the solution

Thus, ideally, a and e should be replaced by this correction factor.

At low concentrations, h is essentially constant, and there is very little variation in

the absorbance, A, with varying concentration of the solution. At high
concentration (> 0.01 M), h may vary significantly and the absorbance (A) is not
directly proportional to concentration. Again, this leads to bending of the
calibration curve.
ii) Intermolecular Effects
As the concentrations of the analyte specie or other species in solution increase,
intermolecular and/or interionic interactions may alter the ability of the analyte
specie to absorb radiation at a given wavelength. Charge distribution changes
(distortion of the molecular energy orbitals) will lead changes in the absorbance
of the analyte species. This effect is not significant below concentrations of ~
0.01 M. Again, bending of the calibration curve occurs.

3. Chemical Interferences
Deviations from Beers law occur when the absorbing specie associates, dissociates
or reacts with the solvent and/or other species in solution, leading to changes in the
concentration of the absorbing specie.
Examples:
i) chromate-dichromate equilibrium

ii) indicator equilibrium

Isobestic Point
Measurements are carried out at the isobestic point, where e1 = e2
Temperature & Equilibrium
In many systems the equilibria are often temperature dependent. Thus changes in
temperature will affect the absorbance of the particular species.
Increases in temperature also shift absorbance bands to longer wavelengths, thus
the absorbances of the species change, also.

4. Spectral Interferences
Spectral interferences are due to many causes.
Stray radiation
Radiation of the same or different wavelength(s) may enter the detector. This
radiation is often referred to as stray radiation and its effect is due to this unwanted
radiation entering the detector during the measurement of P and Po. The effect of
such radiation depends on many factors, including whether the radiation passes
through the wavelength selection device and/or through the sample. The equations
that describe the effects of this unwanted radiation on the absorbance (A) are often
complex and are beyond the scope of this course.
Non-specific molecular absorption
This occurs when concomitant molecular species in the sample absorb radiation in
the same wavelength range as the analyte specie.