Beruflich Dokumente
Kultur Dokumente
Spectroscopy
PHAR 3152
Learning Objectives
By the end of this lecture, the students should be
able to:
1.Describe the visible and ultraviolet (UV) region in
the electromagnetic spectrum.
2.Explain the absorption of radiation.
3.State the reason why compounds appear
colored.
4.Explain electronic transitions and the different
types.
5.State the relationship between Absorbance,
Transmittance and Intensity.
6.Determine unknown concentration using Beers
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Law Plot.
Visible Spectroscopy
A study of interaction of the visible part ( = 380
720 nm) with a chemical species
Main uses:
1. Allows determination of concentration of an
unknown sample.
2. Monitoring of reaction progress as a function of
time.
3. Other quantitative uses.; identification to a
certain extent.
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Visible Light
Each photon has a specific energy; related to certain frequency
or wavelength
380 (blue violet) 700 nm (red)
When all of these wavelength is present, the light appears
white to our eyes
If any wavelength is removed (absorbed), we perceive the
remaining wavelengths of light as the complimentary color
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Colour Wheel
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Chromophore
The nuclei that the electrons hold together in
bonds play an important role in determining which
wavelength of radiation are absorbed
This property of transition and absorption is a
characteristic of a group of atoms not the
electrons themselves
These group of atoms are called chromophores
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Substituents
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How is it measured ?
Usually measures absorbance in liquid solutions
The amount of a particular energy passing
(transmitted) through that solution is quantified
as transmittance
Solvent Cutoffs
Acetonitrile
190 nm
n-Hexane
201 nm
Chloroform
240
Methanol
205
Cyclohexane
195
Isooctane
195
1,4-Dioxane
215
Water
190
95% Ethanol
205
Trimethyl
phosphate
210
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Spectrometer or
Spectrophotometer
1. A light source
2. A diffraction grating
or prism to split light
into different
wavelengths
3. A sample holder or
cell
4. A photosensitive
detector which
measures the light
passing through the
sample
5. An amplifier and an
Absorbance Spectrum
A plot of the
absorbance of a
sample as a function
of wavelength
Can be used to identify
an unknown sample,
since compounds have
characteristic
absorption spectra
Maximum absorption = ~
560 nm
Example
Interferences
Path length
Other absorbing species in the solution
Impurities
Please Watch
By RSC
http://www.youtube.com/watch?v=O
39avevqndU&playnext=1&list=PL44C
6193128D03E4D
By khanacademy.org
http://www.youtube.com/watch?v=qbC
ZbP6_j48&feature=related
Application Question 1
Assay Example 1: Furosemide (frusemide) in tablet form
A typical example of a straightforward tablet assay is the analysis of furosemide (frusemide)
tablets:
1.Tablet powder containing ca 0.25 g of furosemide (frusemide) is shaken with 300 ml of 0.1 M
NaOH to extract the acidic furosemide (frusemide).
2.The extract is then made up to 500 ml with 0.1 NaOH.
3.A portion of the extract is filtered and 5 ml of the filtrate is made up to 250 ml with 0.1 M NaOH.
4.A portion of the extract is filtered and 5 ml of the filtrate is made up to 250 ml with 0.1 M NaOH.
5.The absorbance of the diluted extract is measured at 271 nm.
6.The A (1%, 1 cm) value at 271 nm is 580 in basic solution.
From the data below, calculate the % of stated content in a sample of furosemide (frusemide)
tablets:
Stated content per tablet: 40 mg of furosemide (frusemide)
Weight of 20 tablets : 1.656 g
Weight of tablet powder taken for assay : 0.5195 g
Absorbance reading : 0.596
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Application Question 2
Calculate the percentage of stated content of promazine hydrochloride
in promazine tablets from the following information:
(i)Tablet powder containing ca 80 mg of promazine hydrochloride is
ground to a paste with 10ml of 2 M HCl.
(ii)The paste is then diluted with 200 ml of water, shaken for 15 min
and finally made up to 500 ml.
(iii)A portion of the extract is filtered.
(iv)5 ml of the filtrate is taken and diluted to 100 ml with 0.1 m HCl.
(v)The absorbance is read at a wavelength of 251 nm.
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