definition : a spectrophotometer is an instrument that measure the amount of light absorbed or...
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Definition :Definition :
• A spectrophotometer is an instrument that
measure the amount of light absorbed or
transmitted by the sample.
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Purpose :Purpose :
• Spectrophotometer is used to:1)measure the concentration of the
solution.
2)identify organic compounds by determining the absorption maximum.
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Principle:Principle:
• Spectrophotometer consists of two instruments:
1-spectrometer to produce light for any selected wave length.
2-photometer to measure the intensity of light, and the analyte is put between them.
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SingleSingle andand doubledouble beamsbeams ::
• In the early days of spectroscopy, double beam
spectrophotometers were popular but now it is
thought that the single beam
spectrophotometer is more advantageous.
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Components of single beam :Components of single beam :
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1- Light source1- Light source..
• For ultraviolet absorption spectrophotometer • for visible absorption spectrophotometer
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For ultraviolet absorption For ultraviolet absorption spectrophotometer we use spectrophotometer we use
• # H2 lamp its wave length ranges from 190 to380 nm.
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• #D2 (deuterium) lamp its wave length ranges
from185 to400 nm.
We prefer D2lamp because of its higher stability & it emits continuous radiation.
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for visible absorption spectrophotometer we for visible absorption spectrophotometer we useuse
• #tungsten lamp (350~2200nm)• #tungsten halogen lamp (240~2500nm)
We prefer tungsten halogen lamp because it has longer life, can be used at lower wave
length.
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2-2- Prism or diffraction grating :Prism or diffraction grating : Dispersion devices causes a different
wavelength of light to be dispersion at different angle
Optical materials : lenses.
prism diffraction grating
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3- slit :3- slit : monochromators used for selecting one wave
length .
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4- cell (cuvette):4- cell (cuvette): The cuvette or absorption cells, must made
from material that is transparent to radiation in the spectral region of interest.
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5- Detector (photometer):5- Detector (photometer):
a device used to convert the radiant energy to
electrical signal.
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6- Read out device (Digital galvanometer) :6- Read out device (Digital galvanometer) : The data from the detector are displayed by a
readout device , such as an analog meter ,digital display or liquid crystal display.
The out put can also transmitted to computer or printer .
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Theoretical work of spectrophotometer:Theoretical work of spectrophotometer:
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Used laws:Used laws:
&&
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Where
TT: transmittance of solution. II: intensity of transmitted light . (Watt/cm2)
IIoo: intensity of incident light . (watt/cm2)𝛆𝛆 : Coefficient absorptivity. (L/mole.cm) 𝐜𝐜 : Concentration of solution. (Mole/L): 𝐋: 𝐋Thickness of cuvette. (cm) AA: the absorption of light by the sample
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Beer's Law:Beer's Law:• Beer's Law explains the relationship between
absorbance, at a given wavelength and concentration.
A = C L𝛆
Where:A = absorbance. 𝛆 = molar absorptivity coefficient. (L/mole.cm)L = length of the light path. (cm)C = concentration of the solute. (mole/L)
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Double Beam Spectrophotometer Double Beam Spectrophotometer It is used to measure the ratio of light intensities
on two separate light paths.
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Advantages :Advantages :
1)1) Wide applicability. Wide applicability.
2) High sensitivity.High sensitivity.
3)3) Good accuracy.Good accuracy.
4)4) Ease and convenience.Ease and convenience.
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Disadvantages :Disadvantages :
1. Error in reading due to1. Error in reading due to
»the change in temperature
» nature of solution.
22. It is subject to false wavelength settingIt is subject to false wavelength setting
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33. an error in wavelength setting of ±0.3 nm an error in wavelength setting of ±0.3 nm
results in an error.results in an error.
4. The equipment is generally expensive.4. The equipment is generally expensive.
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Applications in our life Applications in our life
• Clinical• Food and Drink• Industrial / Pharmaceutical• Life Sciences
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Mind mappingMind mapping
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The EndThe End
‘‘ ‘‘best wishesbest wishes ’’ ’’