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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