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Title:Assay of Metronidazole and Diloxanide furoate in

tablet dosage form using a method Absorbance
corrected for interference.

Apparatus required:
1)A double beam UV/Vis Spectrophotometer , 2)Volumetric flasks. 3)Pipettes ,4)Funnels, 5)Filter papers ,
6)Spatula,7)Watch glass or butter paper .

Chemicals required:
1)Metronidazole/Diloxanide furoate tablet, 2)Metronidazole WS, 3)Diloxanide furoate WS, 4)Methanol.
Theory:
UV/Vis Spectrophotometry is the measurement of the absorption of monochromatic radiation by solutions of
chemical substances in the range of 180nm to 380nm (uv) and 380 to 780nm (Vis.) of the spectrum respectively.
According to the modified Beer-Lambert equation

A = log
10
1/T = log
10
I
0
/I = a
1%
1cm
* b * c
Where, T=Transmittance
A=Absorbance which is logarithm of to the base of the 1/T.
I
0
=Intensity of the incident light
I=Intensity of the transmitted light
a
1%
1cm=
Specific absorbance measured in 1%w/v

at 1cm pathlength ,that it is at a particular wavelength in a given
solvent is the property of an absorbing substance.
2

b= Pathlength in cm.
c=Concentration in %w/v.
General calculation techniques for quantitative analysis of a substance in sample:
1)By calibration curve.
2)By using provided A
1%
1cm.
3)By single point standardization technique or direct comparison technique.
Calculation formula in single point standardization technique:
Amount of substance in each unit of a dosage form= A
s8
* C
std
*D.F.*Wt. or Vol. of each unit*Potency of RS
A
std
C
sp
100
The pharmaceutical analyst frequently encounters the situation where the concentration of one or more substances
is required in samples known to contain other absorbing substances which potentially interfere in the assay. In the
many instances substances which are required to be analysed interfere in the assay of each other.
If the identity, concentration and absorbtivity of the absorbing interferents( even substances to be estimated) are
known, it is possible to calculate their contribution to the total absorbance of a mixture. The concentration of the
absorbing component of interest is then calculated from the corrected absorbance (total absorbance minus the
absorbance of the interfering substances) in the usual way,
Procedure:
Each tablet contains Metronidazole = , Diloxanide furoate =
Wt. of 20 tablets=
Avg. Wt. of a tablet=
Grind the tablets to fine powder.

Preparation of Sample Solution:
Weigh well mixed content of 20 tablets equivalent to 45mg of diloxanide furoate, shake to dissolve in methanol.
Make volume up to 100ml with the same solvent. Filter, pipet 10ml and dilute to 50ml with methanol, further
dilute 5ml to 50ml with the same solvent.

Preparation of Standard Solutions:
a)Tinidazole:
Weigh accurately about 0.040g of Metronidazole working standard. Shake to dissolve in methanol and make
volume up to 50ml with the same solvent. Pipet 5ml, dilute to 50ml and further dilute 5ml to 50ml with the same
solvent.

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b)Diloxanide furoate:
Weigh accurately about 0.045g of diloxanide furoate working standard. Shake to dissolve in methanol and make volume up
to 50ml with the same solvent. Pipet 5ml, dilute to 50ml and further dilute 5ml to 50ml with the same solvent.

Measure the absorbance of the sample and standard solutions at 311nm and 257nm.

Observation:
Compounds

Abs.
At 257nm
Abs.
At 311nm
Metronidazole(Std.)
Diloxanide furoate(Std.)
Sample

Calculation:
Compounds

A
1%
1cm
At 257nm
A
1%
1cm
At 311nm
Metronidazole(Std.) X
1
= X
2
=


For Metronidazole:
Metronidazole/Tablet ( Use absorbance at 311nm ):
Abs. of Sp. X Wt. of Std. X 5 X 5 X 100 X 50 X 50 Avg.Wt.X % Potency of standard
Abs. of Std.X Wt. of Sp. X 50 X 50 X 50 X 10 X 5 100

=

For Diloxanide furoate:
Concentration of Metronidazole in the final solution of the assay, C
m
(%) = Samp. Abs. at 311nm =
X
2


=
Contribution made by Metronidazole in the total absorbance at 257nm( Abs
257
)= X
1
* C
m
(%).=
=
Diloxanide furoate/Tablet
=

Abs. of Sp( T.A. at 257nm.- Abs
257
) X Wt. of Std. X 5 X 5 X 100X 50 X 50 Avg.Wt.X % Potency of std.
Abs. of Std. X Wt. of Sp X 50 X 50 X 50 X 10 X 5 100

=

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Metronid azole Diloxanide furoate