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practical

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13 views

1st practical (1)

practical

Uploaded by

paruldamor666
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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AIM: Analysis of pharmacopoeial compound(Paracetamol) by UV Vis

spectrophotometer.
REFERENCE:
REQUIREMENT:
APPARATUS-UV Spectrophotometer, Cuvette, Beaker, Volumetric Flasks
CHEMICAL-Paracetamol,0.1 N NaOH, Methanol, Distilled water.

PRINCIPAL: In UV Spectroscopy any molecule either n, π or σ combination of these


electrons. These bonding (σ & π) and nonbonding (n) electron absorbs the characteristics
radiation and undergo transition from ground state to excited state.
It is a metastable stage where they have a higher energy and lower stability, so they can move
back to the ground state and absorbs the energy as per the nature of electron present and
hence the molecular structure can be elucidated.
BEER-LAMBERT LAW
Beer's law states that "The intensity of a beam of monochromatic light decreases
exponentially with theincrease in the concentration of absorbing species".
Lambert's law states that "The rate of decreases of intensity (monochromatic light) with the
thickness of the medium is directly proportional to the intensity of incident light".
• Beer's Lambert equation,A=abc Where, A=Absorbance, a=Molecular extinction
coefficient, c=concentration(gm/100ml), b=path length(1cm)

THEORY:
Assay-Its an investigative or analytic procedure for assessing or measuring the presence,
amount, or functional activity of a drug (the analyte).
Paracetamol- It is a non-opioid analgesic and antipyretic agent used to treat fever and mild to
moderate pain. It is a widely used over-the-counter medication.
IUPAC ID: N-(4-hydroxyphenyl)acetamide, N-(4-hydroxyphenyl)ethanamide
Other names: N-acetyl-para-aminophenol (APAP), acetaminophen (USAN US)
Formula: C8H9NO2
Molar mass: 151.163 g/mol
Structure of Paracetamol:
Assay using UV-VIS spectroscopy:
The assay of single component sample, which contains other absorbing substances, is then
calculated from the measured absorbance by using one of three following principal
procedures:
1.Use of standard absorptivity value: In standard absorptive value method, the use of
standard A (1%, 1cm) or E values are used in order to determine its absorptivity. It is
advantageous in situations where it is difficult or expensive to obtain a sample of the
reference substance.
2.Calibration graph: In calibration graph method, the absorbances of a number of standard
solutions of the reference substance at concentrations encompassing the sample
concentrations are measured and a calibration graph is constructed. The concentration of
the analyte in the sample solution is read from the graph as the concentration
corresponding to the absorbance of the solution.
3.Single or Double point standardization: The single point standardization procedure
involves the measurement of the absorbance of a sample solution and of a standard
solution of the reference substance. The concentration of the substances in the sample is
calculated from the proportional relationship that exists between absorbance and
concentration.
Ctest= (Atest×Cstd)/Astd

PROCEDURE:
1.By using standard absorptivity value as IP:
I. Weigh and powder 20 tablets.
II. Weigh a quantity of the powder containing about 0.15 g of Paracetamol
III. Add 50 ml of 0.1 M sodium hydroxide, dilute with 100 ml of water, shake for
15 minutes and add sufficient wafer to produce 200.0 ml.
IV. Mix, filter and dilute 10.0 ml of the solution to 100.0 ml with water.
V. To 10.0 ml of the solution add 10 ml of 0.1 M sodium hydroxide,dilute to
100.0 ml with water and mix.
VI. Measure the absorbance of the resulting solution at the maximum at about
257 nm
VII. Calculate the content of C8H9NO2 taking 715 as the specific absorbance at
257 nm.
A=A(1 percent, 1 cm)bc

2.By using calibration curve method:


I. Diluent Preparation: Methanol and Water (15:85, v/v) used as a diluents.
 Standard Preparation: 10mg drug was dissolved in 15ml methanol and was
shaked well. Then 85ml water was added to it to adjust the volume upto
100ml (100ppm). This stock solution is used for further dilution.
II. Test Preparation:
 20 tablets were weighed and powdered.
 Powdered tablet equivalent to 100mg of paracetamol was weighed and taken
into 100ml volumetric flask then 15ml of methanol was added and shaked
well to dissolve it.
 After that 85ml of water was added to adjust the volume upto 100ml .
 From that 1ml of solution was withdrawn and taken in 100ml volumetric
flask.
 The volume was adjusted with diluent up to 100ml.
III. Measure the absorbance of the resulting solution at the maximum at about 243 nm
IV. Calculate the content of PCM from calibration graph.

RESULT:

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