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 Anthracene 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 oxide anion production. It consequently reduces collagenase production in the intra-articular cartilage which spontaneously occurs in the body during destructive inflammation.^{2, 3}

Fig 1. Chemical structures of diacerin (a) and rhein (b)

Literature survey reveals that several spectroscopic and chromatographic methods for analysis of Diacerein in different dosage form like U.V^4-12, Visble^13-15, HPLC^16-22,RP-HPLC^{23-26 ,}LC-MS²⁷, HPTLC²⁸ 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.

· Phosphate Buffer (pH 6.8):- It was prepared according to I.P.1996

· Standard Stock Solution: Accurately weighed (100 mg) pure drug sample of Diacerein was transferred to 100 ml (1000 µg/ml ) calibrated volumetric flask, dissolved and made up to the mark with Phosphate Buffer pH 6.8.

· Developed Method

Different aliquots (0.5, 1.0, 1.5 2.0, 2.5 and 3.00 ml) of a standard Diacerein (10 μg/ml) solution were transferred into a series of 100 ml calibrated flasks and all were made up to the mark with Phosphate Buffer pH 6.8. and absorbance was measured at 258 nm against blank. A calibration curve was plotted from the absorbance values so obtained. A representative spectra and calibration curve of Diacerein is reported in fig.2 and fig. 4 respectively.

Fig. 2: Spectra of Diacerein (pure drug) in Phosphate buffer pH 6.8

Fig. 3: Spectra of Diacerein Capsule (20 µg/ml) formulation

Analysis of Capsule Formulation

Twenty Capsules were weighed accurately and ground into fine powder. An amount of the powder equivalent to 50 mg of Diacerein was weighed and dissolved in about 50 ml Phosphate Buffer pH 6.8. The solution was shaken thoroughly for about 15-20 min, and filtered using Whatman No. 41 filter paper; residue was washed with 20 ml Phosphate Buffer pH 6.8. Filtrate and washing were transferred to a 100 ml calibrated volumetric flask and Phosphate buffer was added up to the mark (500 µg/ml). The 4 ml of above filtrate was diluted to 100 ml with Phosphate buffer pH 6.8. Absorbance was measured at 258 nm wavelength maxima and the concentration of the drug in sample solution was determined from calibration curve. The spectra of formulation and results of analysis are presented in fig.3 and Table 2 respectively.

Fig. 4: Calibration curve of Diacerein (pure drug) in phosphate buffer pH 6.8

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.

Table 1: Quantitative parameters of spectrophotometric method

Parameters	Method
λ_max,nm	258.00
Beer’s law limits, µg/ml	5-30
Molar absorptivity, l mol^-1 cm^-1	23.53 x 10^-3
Regression equation	y = 0.069x + 0.011
Slope	0.069
Intercept	0.011
Correlation coefficient (r²)	0.998

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 Diacerein from Capsule formulation.

RESULT AND DISCUSSION:

One U.V. method has been developed for the quantitative estimation of Diacerein from Capsule formulation. The developed method are based on the solubility of Diacerein in Phosphate buffer having pH 6.8.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 method was found to be simple, accurate and economical hence can be used for routine analysis of Diacerein from pharmaceuticals.

REFERENCES:

1. Harry Gouvas: Use of Sodium Hyaluronate in the treatment of Osteoarthritis, Greece, 2011 Dilip Kumar Renapurkar, Shobana Mathur and K. Jagdishwer Rao, Evaluation of efficacy and safety of diacerein in osteoarthritis of knee joint, Jul-Sep.2010,Vol.1(3)

2. Han-Chun Yao, Xiao-Feng Yang, Sensitive Determination of Nanogram Levels of Diacerein in a Pharmaceutical Formulation by Flow Injection Chemiluminescence Analysis, Journal of the Chinese Chemical Society, 2007, 54, 949-956

3. http://en.wikipedia.org/wiki/Diacerein

4. G. Abirami, K. Anandakumar, R. Velmurugan, Development and Validation of UV-Spectroscopy Method for the Determination of Diacerein Hydrochloride in Pharmaceutical Formulation, Journal of Pharmacy Research, 2012; 5(4), 1949-1951.

5. D M Dhaduk, B G Rathod, P B Patel, Estimation of Sildenafil Citrate and Diacerein Hydrochloride in their Combined Dosage Form by Validated UV Spectroscopic method, Inventi Rapid: Pharm Analysis & Quality Assurance , Vol. 2012 , Article ID- " Inventi:ppaqa/337/12 "

6. P. Selvam, U. Mahalingam, D. Sridharan, and A. Thenmozhi. A Simple UV Spectrophotometric Determination of Diacerein In Pure and In Pharmaceutical Dosage Form, Asian Journal of Biochemical and Pharmaceutical Research, Vol. 1,( 3) 2011

7. Sandeepa Bhoir, Shashikant Dhole, Nilesh Kulkarni, Prajakta Sangole, Sonal Thorat, Dhanashree Bhoite, Novel and validated spectrophotometric estimation of diacerein in bulk and capsule formulation using mixed hydrotropic solubilisation approach, International Journal of Pharmacy and Pharmaceutical Sciences, Vol 4, Suppl 4, 2012

8. Sohan S. Chitlange, Ganesh R. Pawbake, Amir I. Mulla, Sagar B. Wankhede, Simultaneous Spectrophotometric Estimation of Diacerein and Aceclofenac in tablet dosage form, Der Pharma Chemica, 2010, 2(1): 335-341.

9. Krishna R Gupta, Varun E Samrit, Vivek S Thakur and A. T. Hemke, UV-Spectrophotometric estimation of Diacerein in pharmaceutical formulation, Journal of Chemical and Pharmaceutical Research, 2010, 2(3):467-472

10. Narendra Kumar Nyola and Naresh Kalra, Spectrophotometric determination of Diacerein in bulk and pharmaceutical formulation, International Journal of Pharma and Bio Sciences, Oct-Dec.2010,Vol.1(4)

11. Ravindra Pandey, P.O. Patil, M.U. Patil, P.K. Deshmukh, S.B. Bari, Quantitative estimation of Diacerein in bulk and capsule formulation using mixed hydrotropic solubilisation by UV spectroscopy and first order derivative method using area under curve method, Pharmaceutical Methods, Jan-June 2012,Vol 3 (1),4-8

12. K.R .Sreejith,. K. Premalatha, Novel Spectrophotometric Methods for Estimation of Diacerein from Formulations, International Journal of Research in Pharmaceutical and Biomedical Sciences, Vol. 2 (3) Jul – Sep 2011,992-999

13. R Sivakumar, PK Nallasivan, KC Saranya, WD Sam Solomon, T Akelesh, R Venkatnarayanan Visible spectrophotometric estimation of diacerein in bulk and pharmaceutical dosage forms, Pharm Analysis, Year : 2010 ,Volume : 2 ,(4) Page : 414-41

14. R Sivakumar, PK Nallasivan, KC Saranya, Solomon WD Sam, T Akelesh, and R Venkatnarayanan, Visible Spectrophotometric Estimation of Diacerein in Bulk and Pharmaceutical Dosage Forms, J Young Pharm. 2010 Oct-Dec; 2(4): 414–416

15. Sanjay Bais, Indrajeet Singhvi. Novel Colorimetric Methods for quantitative estimation of Diacerin from capsule Formulation. Asian J. Pharm. Ana. 2013; Vol. 3: Issue 2, 44-47.

16. Abha D. Giri, Vidhya K. Bhusari, Sunil R Dhaneshwar, Validated HPLC Method for Simultaneous Quantitation of Tadalafil and Diacerein Hydrochloride in Bulk Drug and Formulation,, International Journal of Pharmacy and Pharmaceutical Sciences,2012;4(2):654-658.

17. S. P. Gandhi, M. G. Dewani, T. C. Borole and M. C. Damle, Development and validation of stability Indicating HPLC method for determination of Diacerein and aceclofenac as bulk drug and In tablet dosage form, International Journal of Research in Pharmacy and Chemistry, 2011, 1(4).

18. Keddal G. Lalithaa, Thangavel Venkatachalamb, Ramaraj Srinivasanb, Ponnusamy Kalaiselvib, Nagappan Kannappan, A Simple HPLC Method for Quantitation of Diacerein in Tablet Dosage Form, Eurasian J. Anal. Chem. 5(1): 81-88, 2010

19. Krishna R Gupta, Varun E. Samrit, Anuradha H Mahapatra, Sudhir G. Wadodkar, Stability indicating HPLC assay method for Diacerein and Aceclofenac in tablets, 2011Volume 1, (3).

20. Rao J, Chauhan K, Mahadik KR, Kadam SS. A stability-indicating high performance liquid chromatographic method for the determination of diacerein in capsules. Indian J Pharm Sci 2009; 71:24-9.

21. S. P. Gandhi, M. G. Dewani, T. C. Borole and M. C. Damle, Development and validation of stability indicating HPLC method for determination of Diacerein and Aceclofenac as bulk drug and in tablet dosage form, International Journal of Research in Pharmacy and Chemistry, 2011, 1(4),779-806.

22. Keddal G. Lalithaa, Thangavel Venkatachalamb, Ramaraj Srinivasanb, Ponnusamy Kalaiselvib, Nagappan Kannappanc, A Simple HPLC Method for Quantitation of Diacerein in Tablet Dosage Form, Eurasian J. Anal. Chem. 5(1): 81-88, 2010.

23. KS Rao, NK Keshar, J Datta, Stability indicating RP-HPLC method for the simultaneous estimation of Diacerein and Aceclofenac in bulk and pharmaceutical dosage forms Inventi Impact: Pharm Analysis & Quality Assurance, Vol. 2010.

24. D. Kiran Kumar, K.S. Nataraj, K. Kesinath Reddy, S. Suresh Kumar, Method Development and Validation for analysis of Diacerein in Bulk and Pharmaceutical Dosage Form by RP-HPLC.

25. N. Kannappan, Madhukar A., R. Srinivasan, R.L.A. Srinivas , C H. Naveen Kumar, Mannavalan. R, Analytical method development and validation of Diacerein Tablets by RP-HPLC, Vol.2, No.1, Jan-Mar 2010, 143-148.

26. Narendra Nyola, Govinda Samy Jeyabalan, N Kalra, Gazala Parveen, and Suresh Choudhary, Development and Validation of a RP-HPLC Method for Simultaneous Estimation of Diacerein and Aceclofenac in Pharmaceutical Dosage Form, Research and Reviews: Journal of Pharmaceutical Analysis, Oct–Dec, 2012, Vol 1 (1).

27. Arun Shirwaikar, A. Sarala Devi, K Premalatha and KR. Sreejith, Determination of Diacerein in Rabbit Plasma by Liquid Chromatography - Mass Spectroscopy and Its Application to Bioavailability Study, International Journal of Research in Pharmaceutical and Biomedical Sciences, Oct – Dec 2012,Vol. 3 (4),1738-1744.

28. S. P. Gandhi, M. G. Dewani, T. C. Borole, and M. C. Damle,Development and Validation of Stability Indicating HPTLC Method for Determination of Diacerein and Aceclofenac as Bulk Drug and in Tablet Dosage Form, E-Journal of Chemistry, 2012, 9(4), 2023-2028.

Received on 29.10.2013 Accepted on 01.12.2013

Asian J. Pharm. Ana. 3(4): Oct. - Dec. 2013; Page 124-126

Method	Brand	Label Claim (mg/Capsule)	%Label Claim Estimated*	S.D.	% Recovery**
U.V	A	50	99.71	0.164	98.99
U.V	B	50	99.17	0.327	99.25