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ESTIMATION OF QUININE SULPHATE BY FLUORIMETRY (Practical IMA 2024)

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355 views3 pages

ESTIMATION OF QUININE SULPHATE BY FLUORIMETRY (Practical IMA 2024)

Uploaded by

himanshukurrey951
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© © All Rights Reserved
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5. ESTIMATION OF QUININE SULPHATE BY FLUORIMETRY.

Aim: To estimate the amount of Quinine sulphate present in given sample by fluorimetry.
Requirements:

Apparatus: Pipette, volumetric flask, measuring cylinder.


Chemicals: Quinine sulphate , 0.1N H2SO4 (Sulphuric acid), Distilled water.
Instrument: Fluorometer
Reference:

Principle:
Fluorescence is the emission of visible light by a substance that has absorbed light of a different
wavelength. The emitted photon has a longer wavelength and lower energy. As the excitation
of the molecule is due to the absorption of a photon (light), these types of luminescence are
called photoluminescence. Fluorescence is also defined as the radiation emitted in the transition
of a molecule from a singlet excited state to a singlet ground state.

The wavelength of absorbed radiation is called as excitation wavelength and that of emitted
radiation is called as emission wavelength. Fluorescent intensity is directly proportional to
concentration of substance. Fluorometer is the instrument used to measure fluorescence with a
wavelength varying between 240 nm to 800 nm. Such measurement form the basis of a
sensitive method of analysis called the fluorometry. Fluorometric methods of analysis have
found application in many situations of pharmaceutical interest such as in analysis of
riboflavin, thiamine,reserpine, quinine sulphate in drug dosage forms.
Theory:
Absorption of UV/Visible radiation causes transition of electrons from singlet ground state to
singlet excited state. As this state is not stable, it emits the energy in the form of UV/Visible
radiation and returns to singlet ground state. This study or measurement of this emitted
radiation (when electrons undergo transition from singlet excited state to singlet ground state)
is the principle in Flourimetry. Phosphorescence is also a related phenomenon, which is the
study of emitted radiation when electrons undergo transition from triplet state to singlet ground
state.

Procedure:
Preparation of 0.1 N H2SO4:-
Pipette out 2.7 ml of conc H2SO4 to 100 ml of water and then make it up to 1000 ml with
water.
Preparation of standard Quinine sulphate solution
 Weigh accurately 100mg of Quinine sulphate powdered drug
 Dissolve in 100 ml of 0.1 N conc. H2SO4 to get 1mg/ml
 Take 10 ml of above solution and dilute to100 ml with 0.1N H2SO4(100μg/ml)
 Again 10ml of above solution dilute to 100ml with 0.1N H2SO4(10μg/ml)
 Pipette out 0.5,1, 1.5, 2,2.5,3 ml of above diluted standard quinine sulphate solution in to
a set of 10 ml volumetric flask and make up to 10 ml with 0.1N H2SO4 to get
concentration of 0.5,1,1.5,2,2.5,3 µg/ml respectively.
 Switch on the instrument and stabilize for 10-15min.
 Set excitation and emission filters at the wavelengths 365 and 459nm respectively.
 Set the fluorescence intensity to 0% by using 0.1N H2SO4 as blank and 100% by using
highest concentration of the standard solution. Measure the fluorescence of serial
dilutions and plot the calibration curve (fluorescence intensity Vs concentration).
Observation:
S. No. Concentration (µg/ml) % Fluoroscence intensity
1. 0.5
2. 1.0
3. 1.5
4. 2.0
5. 2.5
6. 3.0
7. Unknown

Report: The amount of Quinine sulphate present in the given sample was found to be ……..

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