Application Of UV Visible Spectroscopy In The

Separating Process:
Submitted to : Prof. Amir Iqbal
Submitted by:
Name : Atta Subhan
Muhammad Husnain
Roll no. : 364704
364705
Course Title : Analytical Chemistry-II
Course Code : CHM-508
Program : BS Chemistry 6th Semester
(Evening)
Govt. Graduate College Samanabad, Faisalabad.
 Introduction:

 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 color 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 1,3,5-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.
THANK YOU