Application of UV visible spectroscopy in the separating process:

UV visible spectroscopy is routinely, used in analytical chemistry for the quantitative

determination of different analytes, such as transition metal ions, highly conjugated organic
compounds, and biological macromolecules. Spectroscopic analysis is commonly, carried out in
solutions but solids and gases may also be studied, identified, and separated based on absorption
in the UV visible region of the spectrum.

Differentiation between copper sulfate and ammonical copper sulfate:

Solutions of transition metal ions can be colored (i.e. absorb visible light) because d-electrons
within the metal atoms can be excited from one electronic state to another.

The color of metal ions solutions is strongly affected by the presence of other species such as
certain ions or ligands. For instance, the color of a dilute solution of copper sulfate is a very light
blue; adding ammonia intensifies the cooler and changes the wavelength of maximum
absorption. In this way, copper sulfate and ammonical copper sulfate can be separated by UV-
visible spectroscopy.

The separation between different concentrations of the same or different substances:

The Beer-Lambert law states that the absorbance of a solution is directly proportional to the
concentration of the absorbing species in the solution and the path length.

For a fixed path length, UV visible spectroscopy can be used to determine the concentration of
the absorber in a solution. It is necessary to know how quickly the absorbance changes with
concentration.

This can be taken from references (tables of molar extinction coefficients), or more accurately
determined from a calibration curve.

So the different concentrations of the same or different substances can be identified and
separated based on their absorbance max.

Identification of different conjugated systems:

UV visible spectroscopy is helpful in the separation of different conjugated systems.

A simple carbon-carbon double bond (for example in ethene) has a maximum absorption at 171
nm. The two conjugated double bonds in but-1,3-diene have a maximum absorption at a longer
wavelength of 217 nm while for 1,35-hexatriene absorption occurs at a still longer wavelength
and absorbs at 258 nm.

Thus progressively increasing wavelength moves the absorption to longer wavelengths and
finally into the visible region.

Polyenes having a conjugated system of 20 double bonds are yellow in color.

The red color of carrots and tomatoes arises from the conjugated molecule of this type.

Identification/separation of functional groups:

The wavelengths of absorption peaks can be correlated with the types of bonds in a given
molecule and are valuable in determining or separating the functional groups within a molecule.

The shift in absorption in these cases results from a combination of inductive and resonance
effects. Substitutions may change the energy level of the both the ground and excited state but
the important point is the relative energies of the two states.

Analysis/separation of multicomponent mixtures:

It is relatively rare to find a practical problem in which one has a mixture to be analyzed with
only one component, which absorbs radiation.

When there are several such components, which absorb at the same wavelength their absorbance,
adds together, and it is no longer true that the absorbance of the sample is proportional to the
concentration of the component.

In these cases, several approaches can be adopted with the most important being chemical
reaction and multi-wavelength measurements.

Multicomponent analyses using UV-Visible spectra have been performed for almost as long as
single-component analyses, but because the techniques used in multicomponent analysis often
gave incorrect results (as detailed below), they were not widely applied.

However, modern instruments yield more precise data, and modern curve-fitting techniques give
more accurate results and perhaps more importantly indicate when results are incorrect. For these
reasons, multicomponent UV visible analyses are becoming more popular.

Separation in analytical chemistry:

Since the band of UV visible is, intense so small quantity of the substance can easily be detected.
For example, the presence of benzene in cyclohexane can be detected easily by using the
spectroscopic technique.
Benzene exhibits very strong light absorption near 180 nm (E > 65,000), weaker absorption at
200nm (E = 8,000), and a group of much weaker bands at 254 m (E = 240), while cyclohexane
due to the absence of chromophores or conjugated system is inactive to UV visible spectroscopy.

Drug separation: Many drugs in the form of raw material or in the process of formulation can be
separated from the solvent by measuring the absorbance at a specific wavelength.