INTRODUCTION:

Diacerein also known as diacetylrhein, is a drug used in the treatment of osteoarthritis.¹Diacerein is the drug to be proved as disease modifying agent. Diacerein [4,5-bis[acetyloxy]-9,10-dioxo-dioxo-2 –anthracene] is an Anthracine derivative. It is converted to active metabolite “Rhein” which has anti inflammatory effects through inhibition of Interleukin-1B. It reduces the fibrinolytic synovial Fibroblasts. It also dose –dependently inhibits chemotaxis and super oxideanion production. It consequently reduces collagenase production in the intraarticular cartilage which spontaneously occurs in the body during destructive inflammation.^2,3 Literature survey reveals that several spectroscopic and chromatographic methods for analysis of Diacerin in different dosage form like U.V^4-12,Vsisble ^13,14,HPLC^15-22,RPHPLC^23-26LC-MS,²⁷HPTLC ²⁸ were reported.

MATERIALS AND METHODS:

Apparatus

A Shimadzu UV/Visible double beam spectrophotometer (Model 1700) with 1 cm matched quartz cells were used in present study for spectral and absorbance measurements.

Reagents and Materials

All reagents used were of analytical grade and double distilled water was used throughout the investigation.

· Orange II Reagent: Dye solution (0.1% w/v) was prepared in Buffer solution pH 9.2.

· Tropoelin Reagent: Dye solution (0.1% w/v) was prepared in Buffer solution pH 9.2.

· Standard Stock Solution: Accurately weighed (50 mg) pure drug sample of Diacerin was transferred to 50 ml calibrated volumetric flask, dissolved and made up to the mark with chloroform.(1000 µg/ml)

Method A

Different aliquots (0.5, 1.0, 1.5,2.0,2.5 3.0 and 3.5 ml) of a standard Diacerin (1000 μg/ml) solution were transferred into a series of 10 ml calibrated flasks and all were made up to the mark with chloroform. To the above solution, 5 ml of Orange II reagent was added in a separating funnel. Reaction mixture was shaken gently for 5 min and allowed to stand so as to separate the aqueous and chloroform layer. Coloured chloroform layer was separated out and absorbance was measured at 465 nm against reagent blank. A calibration curve was plotted from the absorbance values so obtained. A representative spectra and calibration curve of ion pair complex of Diacerin and Orange II dye is reported in fig 1 and fig 2 respectively.

Method B

Different aliquots (0.5, 1.0, 1.5,2.0and 2.5 ml) of a standard Diacerin (1000 μg/ml) solution were transferred into a series of 10 ml calibrated flasks and all were made up to the mark with chloroform. To the above solution, 5 ml of Tropoelin reagent was added in a separating funnel. Reaction mixture was shaken gently for 5 min and allowed to stand so as to separate the aqueous and chloroform layer. Coloured chloroform layer was separated out and absorbance was measured at 465.0 nm against reagent blank. A calibration curve was plotted from the absorbance values so obtained. A representative spectra and calibration curve of ion pair complex of Diacerin hydrochloride and Tropoelin dye is reported in fig 3 and fig 4 respectively.

Analysis of Capsule Formulation

Twenty Capsules were weighed accurately and ground into fine powder. An amount of the powder equivalent to 50 mg of Diacerin was weighed and dissolved in about 50 ml chloroform. The solution was shaken thoroughly for about 15-20 min, and filtered using Whatman No. 41 filter paper; residue was washed with 20 ml chloroform. Filtrate and washing were transferred to a 100 ml calibrated volumetric flask and chloroform was added up to the mark. Filtrate 1 ml (method A) and 1.0 ml (method B) were diluted to 10 ml with chloroform. These final dilutions were treated as per the procedure for respective calibration curve. Absorbance was measured at respective wavelength maxima and the concentration of the drug in sample solution was determined from calibration curve. Results of analysis are presented in Table 2.

The optical characteristics such as Beer’s law limit, molar absorptivity were calculated and summarized in Table 1. Regression equation, correlation coefficient, slope and intercept are also shown in Table 1.

Recovery Studies

Recovery studies were carried out by addition of pure drug to previously analysed Capsule sample at three different concentration levels. The results of recovery studies are reported in Table 2. The results of recovery studies reflect that there is no interference of excipients in the analysis of Diacerin Hydrochloride from Capsule formulation.

RESULT AND DISCUSSION:

Two colorimetric methods have been developed for the quantitative estimation of Diacerin from Capsule formulation. The developed methods are based on the formation of ion pair complex of drug with Orange II Dye and Tropoelin Dye. The results of analysis from Capsule formulation were within the permissible limits and the results of recovery studies reflect nil interference from excipients. The developed methods were found to be simple, accurate and economical hence can be used for routine analysis of Diacerin Hydrochloride from pharmaceuticals.

Fig. 1: Spectra of ion pair complex of Diacerin HCl and Orange II dye

Fig. 2: Calibration curve of ion pair complex of Diacerin and Orange II dye

Table 1: Quantitative parameters of spectrophotometric method

Parameters	Method A	Method B
λ_max,nm	465.00	465.00
Beer’s law limits, µg/ml	50-350	50-250
Molar absorptivity, l mol^-1 cm^-1	1.7756 X 10⁴	1.334 X 10⁴
Regression equation	y = 0.003x + 0.206	y = 0.002x + 0.254
Slope	0.003	0.002
Intercept	0.206	0.254
Correlation coefficient (r²)	0.991	0.998
SD	0.3527	0.0522

Table 2: Results of Analysis of Capsule Formulation and Recovery Studies

Method	Brand	Label Claim (mg/Capsule)	%Label Claim Estimated*	S.D.	%Recovery**
Using Orange II Dye	A	30	98.71	0.164	98.99
Using Orange II Dye	B	30	99.33	0.327	99.25
Using Tropoelin Dye	A	30	99.43	0.216	99.87
Using Tropoelin Dye	B	30	100.3	0.324	100.20

*Average of six determinations

** Average of Recovery Studies at three different concentration levels

Fig. 3: Spectra of ion pair complex of Diacerin and Tropoelin dye

Fig. 4: Calibration curve of ion pair complex of Diacerin HCl and Tropoelin dye

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Received on 06.04.2013 Accepted on 21.05.2013

Asian J. Pharm. Ana. 3(2): April- June 2013; Page 44-47