INTRODUCTION:

Fenofibrate chemically 1-methylethyl2-[4-(4-chlorobenzoyl) phenoxy]-2-methyl- propionate. Its therapeutic effects through activation of peroxisome proliferators activated receptor a (PPARa). Fenofibric acid, the active metabolite of fenofibrate. Increases Apo lipoprotein A-Iα Mediated High-Density Lipoprotein Biogenesis by enhancing transcription of ATP-Binding cassette transporter A1 gene in a liver X receptor dependent manner. There are varies method are reported for estimation of Fenofibrate in pharmaceutical lick UV-Spectroscopy,^{1, 2}. HPLC^{3, 7-8}, Solid phase extraction coupled to liquid chromatography⁴, UPLC⁵, LC-MC⁶.

Experimental:-

Instrument, Chemicals, Reagents:

Shimadzu 1700 U.V visible spectrophotometer with 1cm matched quartz cells, and methanol was used for the experiment.

Fenofibrate Stock Solution:

25 mg portions of standard Fenofibrate were weighed, transferred to 25 ml volumetric flasks, and dissolved in methanol (15 ml). The flask was shaken, and the volume was made up to the mark with same solvent. A 1.0 ml aliquot was pipetted into a 10 ml volumetric flasks sepretly, and methanol was added to adjust the volume to give a concentration of 100 μg/ml of Fenofibrate respectively.

Sample solution of pharmaceutical formulation (Tablet):

Sample solution: Twenty tablets of Fenofibrate were weighed and powered in glass mortar. Amount equivalent to 10 mg was transferred to 100 ml volumetric flasks, dissolved in sufficient quantity of methanol, sonicated and made up the volume with methanol to obtain concentration of 100µg/ml. This solution was then filtered through Whitman filter paper # 41. Further dilutions were made from this stock solution to get required concentration.

Determination of λ max:

The standard solution of Fenofibrate (12µg/ml) was scanned at different concentrations in the range of 200-400nm and the λ max was found to be 285.5nm against reagent blank.(Fig 1) The absorbances were recorded for 03-30 μg/ml at 285.5nm (λ max). From this calibration curve was plotted. (Fig 2)

Second Order Derivative Method:

The standard solution of Fenofibrate were diluted appropriately with methanol to obtain solutions containing 12 μg/ml. Spectra of these diluted solutions were scanned in the spectrum mode between 200 and 350 nm. These zero-order spectra of Fenofibrate were treated to obtain corresponding second-order derivative spectra (Fig 3)

Calibration Curve for Fenofibrate:

The absorbances were recorded for 03-30 μg/ml at 211 nm in derivative mode. From this calibration curve was plotted. (Fig 4)

Fig :1 λ max for Fenofibrate:

Fig :2 Calibration Curve for Fenofibrate:

Fig :3: second Order Derivative Spectra of Fenofibrate:

Fig :4: Calibration Curve for Fenofibrate

Table:-1 Calibration curve for Fenofibrate:

Sr. No.	Conc. of Fenofibrate	Absorbance at 285.5
1	3	0.150
2	6	0.313
3	9	0.462
4	12	0.627
5	15	0.794
6	18	0.945
7	21	1.103
8	24	1.255
9	27	1.412
10	30	1.569

Table :2- Optical Characteristics and Precision

Following are the parameters obtained during the procedure.

Parameters	Zero Order Values	Second Order Derivative
Absorption maxima (nm)	285.5	217
Beer’s law limit (mcg/ml)	03-30	03-30
Correlation coefficient	0.9998	0.9994
Molar absorptivity (lit/mole/cm)	0.7315×10⁴	1.0183×10⁴
Sandell’s sensitivity(mcg/sqcm/0.001)	0.049322	0.02376
Slope (m)	0.02027	0.0765
Intercept	0.001333	0.016772
% COV	0.099995	0.137648
Confidence limit with 0.05 level	0.001530	0.001434
LOD(ng\ml)	100	90
LOQ(ng\ml)	200	250

Table :3:- Results of analysis of Zero Order

Sr.No	Tablet	Label claim	% Estimated	% Recovery*
1	Marketed tablet A	160	99.46	100.03
2	Marketed tablet B	200	100.122	99.96

Table :4:- Statistical analysis of results

Sr.No	Tablet	SD*	COV (%)*	SE*
1	Marketed tablet A	0.05451	0.05480	0.03147
2	Marketed tablet B	0.08138	0.08120	0.04698

* Mean of three readings

SD-Standard deviation, COV-Coefficient of Variation, SE- Standard Error

Table :5:- Results of analysis of Second Order.

Sr.No	Tablet	Label claim	% Estimated	% Recovery*
1	Marketed tablet A	10	99.77	98.51
2	Marketed tablet B	20	99.97	99.14

Table : 6:-Statistical analysis of results

Sr.No	Tablet	SD*	COV (%)*	SE*
1	Marketed tablet A	1.4246	1.4953	0.8649
2	Marketed tablet B	1.3047	1.3050	0.7512

^*Mean of three readings SD-Standard deviation, COV-Coefficient of Variation, SE- Standard Error

Method validation^9-12:

Specificity-

Pure Fenofibrate is spiked with common excipients and was assayed by proposed method and it was found that the assay results were unaffected by the presence of such excipients.

Linearity-

Linearity was observed in the range of 03-30 mcg/ml for zero order,second order derivative. The calibration curve yielded coefficient of correlation (r) 0.9998, 0.9994, for zero order, second order derivative method respectively.

Sensitivity-

High Molar absorptivity and low Sandell’s sensitivity for the respective method reveals that all these methods are highly sensitive.

System precision-

% COV calculated from 3 replicate readings (absorbance values) at concentration (12 µg/ml) confirm the precision of the method.

Assay results-

Two different brands of Fenofibrate tablets were analysed by proposed methods, the percentage in tablet were determined and presented in the table. Assay results obtained are within limit.

Accuracy and precision –

The low values of S.D, %COV, and 95% confidence interval indicate that method is precise. % recovery was found to be within limit indicate the noninterference from the formulation excipients and confirm the accuracy and precision of the method.

RESULTS AND CONCLUSIONS:

All the methods Zero order, second order derivative method for the estimation of Fenofibrate in tablet dosage were found to be simple (table:-2), accurate and reproducible. Beer- lambert’s law was obeyed in the concentration range of 3-30 µg/ml in all these methods. The accuracy of the method was assessed by recovery studies at three different levels i.e. 80%, 100%, 120%. The values of standard deviation were satisfactory and the recovery studies were close to 100%. The %RSD value is less than 2 indicative of accuracy of the method. Results found are satisfactory.

ACKNOWLEDGEMENTS:

Authors are grateful to Emcure Pharmaceuticals Bhosari, Pune for providing the gift sample of Fenofibrate. We are also thankful to the Principal and Head of Pharmaceutical Chemistry Department of Government College of Pharmacy, Karad for providing the necessary facilities to carry out this work.

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Received on 25.11.2009 Modified on 14.01.2010

Asian J. Research Chem. 3(2): April- June 2010; Page 326-328