UV-Visible Spectroscopy

A diagram of the components of a typical spectrometer are shown in the following

diagram. The functioning of this instrument is relatively straightforward. A beam of
light from a visible and/or UV light source (colored red) is separated into its
component wavelengths by a prism or diffraction grating. Each monochromatic
(single wavelength) beam in turn is split into two equal intensity beams by a halfmirrored device. One beam, the sample beam (colored magenta), passes
through a small transparent container (cuvette) containing a solution of the
compound being studied in a transparent solvent. The other beam, the reference
(colored blue), passes through an identical cuvette containing only the solvent.
The intensities of these light beams are then measured by electronic detectors
and compared. The intensity of the reference beam, which should have suffered
little or no light absorption, is defined as I0. The intensity of the sample beam is
defined as I. Over a short period of time, the spectrometer automatically scans all
the component wavelengths in the manner described. The ultraviolet (UV) region
scanned is normally from 200 to 400 nm, and the visible portion is from 400 to
800 nm
https://www2.chemistry.msu.edu/faculty/reusch/virttxtjml/spectrpy/uvvis/uvspec.htm#uv1
Infrared spectroscopy is an absorption method in the wavelength region of 1 to
100 m in that extends the region of the visible light to longer wavelengths and
smaller frequencies/energies. The energy of infrared light is no longer sufficient
to induce transitions of valence electrons. Instead, infrared radiation excites
vibrational and rotational motions in molecules. Except for the differences in the
energy transfer from the radiation to the molecule, the principles of IR
spectroscopy are the same as those of VIS/UV spectroscopy or other
spectroscopic techniques. The absorption of infrared light is again characterized
by the BougerLambert-Beer Law. However, infrared spect
http://www.uni-konstanz.de/FuF/Bio/folding/Chapter%202_IR.pdf

Principle of UV spectroscopy
UV spectroscopy obeys the Beer-Lambert law, which states that: when a
beam of monochromatic light is passed through a solution of an absorbing
substance, the rate of decrease of intensity of radiation with thickness of
the absorbing solution is proportional to the incident radiation as well as the
concentration of the solution.
The expression of Beer-Lambert law is-

A = log (I0/I) = Ecl

Where, A = absorbance
I0 = intensity of light incident upon sample cell
I = intensity of light leaving sample cell
C = molar concentration of solute
L = length of sample cell (cm.)
E = molar absorptivity
From the Beer-Lambert law it is clear that greater the number of molecules
capable of absorbing light of a given wavelength, the greater the extent of
light absorption. This is the basic principle of UV spectroscopy.
http://www.indiastudychannel.com/resources/146681-Principle-workingand-applications-of-UV-spectroscopy.aspx