Uv Visible Spectroscopy : By:-Sarika Singh. Fy M-Pharmacy (QAT)
Uv Visible Spectroscopy : By:-Sarika Singh. Fy M-Pharmacy (QAT)
SPECTROSCOPY……
by:- Sarika Singh.
Fy M-Pharmacy(QAT)
Spectroscopy
It is the branch of science that deals with the study of
interaction of matter with light.
OR
Frequency (ν):
◦ It is defined as the number of times electrical field
radiation oscillates in one second.
◦ The unit for frequency is Hertz (Hz).
1 Hz = 1 cycle per second
Wavelength (λ):
◦ It is the distance between two nearest parts of the wave in the
same phase i.e. distance between two nearest crest or
troughs.
The relationship between wavelength & frequency can be written as:
c=νλ
E=hν=hc/λ
ABSORBANCE LAWS
BEER’S LAW
mathematically;
-dI/ dt ᾱ I
-In . I = kt+b -------------- eq(2)
E=h𝜗 erg
E₁-Eₒ= h𝜗
Rotational
Electronic Transitions
The possible electronic transitions are
1- σ → σ* transition
2- π → π* transition
3- n → σ* transition
4- n → π* transition
Absorption & Intensity Shifts
Blue Red
Absorbance ( A )
shift shift
Hypochromic shift
λmax Wavelength ( λ )
Instrumenation:
Source of radiation.
Collimating system.
Mono-chromator system.
Sample holder.
Detector.
Amplifier and Read-out devise.
Source of radiation
Requirements of an ideal source
1. Filters
2. Prisms
3. Gratings
1. Filters
1. Diffraction grating.
2. Transmission gratings.
Sample holder or cuvettes
a) They must be uniform in construction, the thickness must be
constant and surfaces facing the incident light must be optically
flat.
b) The materials of construction should be inert to solvents.
c) They must transmit light of the wavelength used.
Detectors
Device which converts light energy into electrical signals, that
are displayed on readout devices.
The transmitted radiation falls on the detector which determines
the
intensity of radiation absorbed by sample.
The following types of detectors are employed in instrumentation
of absorption spectrophotometer
1. Barrier layer cell/Photovoltaic cell
2. Phototubes/ Photo emissive tube
3. Photomultiplier tube
Requirements of an ideal detector:-
a. It should give quantitative response.
b. It should have high sensitivity and low noise level.
c. It should have a short response time.
d. It should provide signal or response quantitative to wide
spectrum of radiation received.
Single beam UV-Spectrophotometer
Double Beam UV-Spectrophotometer
Applications of
UV / Visible
Spectroscopy
1. Detection of Impurities.
4. Qualitative analysis
5. Chemical kinetics
6. Detection of Groups
9. As HPLC Detector