Simultaneous Spectrophotometric Estimation of Diacerein and Aceclofenac by Vierodt’s Method and First Derivative Method
*Corresponding Author E-mail: bsantosh83@yahoo.com
ABSTRACT:
Two novel Spectrophotometric methods, simultaneous equation method (Vierodt’s Method) and first derivative method were developed, validated and compared for the simultaneous estimation of diacerein and aceclofenac in their combined tablet dosage form. In Vierodt’s method, λ max of diacerein and aceclofenac, 257.6 nm and 274.0 nm respectively were selected for estimation. In first derivative spectrophotometric method the zero crossing technique was applied for the estimation of diacerein and aceclofenac in which 250.0 nm and 298.40 nm were selected respectively. The diacerein and aceclofenac follow Beer-Lambert’s law in the concentration ranges from 2.5 to 15µg/mL and 5.0 to 30µg/mL respectively. The correlation coefficient was found to be 0.9998 for diacerein and 0.9998 for aceclofenac by Vierodt’s method. By First Derivative method, correlation coefficient was found to be 0.9999 for diacerein and 0.9985 for aceclofenac. Mean recoveries were found satisfactory. These procedures do not involve any separation step. Proposed methods were successfully applied for the simultaneous estimation of diacerein and aceclofenac in the combined tablet dosage forms.
KEYWORDS: Diacerein, Aceclofenac, Vierodt’s method, Derivative method, Simultaneous
INTRODUCTION:
Combination of diacerein and aceclofenac is frequently prescribed as an analgesic and anti-inflammatory agent in rheumatoid arthritis and other types of joint disorders. Diacerein and aceclofenac combination is recently introduced in the market for its synergetic effect in the treatment of different joint disorders. Diacerein is chemically known as 4, 5-diacetoxy-9-10-dioxo-9-10-dihydroanthracene-2-carboxylic acid. It is a novel anti-inflammatory drug with pharmacological properties different from those of classical non steroidal anti-inflammatory agents. It is a disease modifying anti-rheumatoid drug used in the treatment of Osteoarthritis and chronic inflammatory arthritis. It is also known to inhibit interleukin-1. Aceclofenac ([2-(2´, 6´-dichlorophenylamino) phenyl] acetoxyacetic acid) is a phenyl acetic acid derivative that shows analgesic properties and good tolerability profile in a variety of painful conditions.
It is largely used in the symptomatic treatment of pain and of inflammatory or degenerative arthropathies like osteoarthritis, rheumatoid arthritis and ankylosing spondylities. Literature survey revealed that various methods including stability-indicating[1], chemiluminescence’s analysis [2], isolation and structural elucidation of impurity [3], physico-chemical and structural characterization [4], LC-MS/MS [5], HPLC [6], stability-indicating [7], narrowbore HPLC using column-switching [8], spectrophotometric and spectrofluorimetric [9,10], reverse phase HPLC [11], capillary electrophoresis [12], in human plasma by HPLC [13], are used for the determination of diacerein and aceclofenac alone or in its combination with other drugs. However, we present here two novel UV-Spectrophotometric methods, Vierodt’s Method (Fig.1.Overlay spectrum of diacerein and aceclofenac) and First Derivative method (Fig.2 a, 2 b-Overlay spectrum of diacrein and aceclofenac respectively) for the rapid simultaneous estimation of diacerein and aceclofenac in the combined tablet dosage form. The proposed methods are demonstrated here for routine simultaneous estimation of diacerein and aceclofenac in combined tablet dosage forms.
MATERIALS AND METHODS:
Materials:
Diacerein and aceclofenac were kindly supplied by Blue Cross Laboratories Pvt. Ltd. Nashik, India. Formulation of diacerein and aceclofenac in combined tablet dosage form was purchased from local market. Methanol (HPLC grade) was purchased from Fischer Scientific (India). Dimethylsulfoxide (GR grade) was purchased from Merck Co.; double distilled water.
Fig.1- Overlay Spectrum of diacrein 10 µg/ml and aceclofenac 10 µg/ml
Fig.2 (a)- First derivative overlay spectrum of diacrein 2.5-15 µg/ml
Instrumentation:
Shimadzu UV-2450 double beam spectrophotometer with 1 cm path length, supported by Shimadzu UV-Probe software, version 2.21 was used for all spectrophotometric estimations. Shimadzu balance (AUW-120D) was used for all weighing. Ultrasonicator was used for sonication of all analytical solutions.
Fig.2 (b)- First derivative spectrum of aceclofenac 5-30 µg/ml
Standard stock solution:
Diacerein was dissolved initially in sufficient dimethyl sulfoxide to dissolve the content and further diluted with methanol. This gave concentration of 1mg/mL. Aceclofenac solution of 1mg/mL was prepared by using methanol as solvent. Working standard solutions were further prepared from above standard stock solutions in methanol to give concentration of 100µg/mL and 200µg/mL of diacerein and aceclofenac respectively.
Sample preparation:
Twenty tablets were accurately weighed and powdered. A portion of tablet powder equivalent to 10 mg of diacerein and 20 mg of aceclofenac was accurately weighed and transferred into a 100mL volumetric flask and then added 5mL of dimethylsulfoxide and 10mL of methanol to dissolve contents of tablet formulation. Then, solution was sonicated for 10 minutes and same contents were filtered through Whatman filter paper 41. The final volume was made up to 100mL with HPLC grade methanol to obtain concentration of 100µg/mL and 200µg/mL of diacerein and aceclofenac respectively.
Method validation:
Linearity:
From working solution of 100 mg/mL of diacerein and 200 mg/mL of aceclofenac, 0.25, 0.50, 0.75, 1.0, 1.25, 1.5 mL were transferred into series of 10mL volumetric flasks to obtain concentration range of 2.5 to 15 mg/mL for diacerein and 5 to 30mg/mL for aceclofenac.
Table 1: Results of validation parameters, linearity range, LOD, LOQ, precision (Intra and Inter day)
|
Parameters |
Vierodt’s Method |
First Derivative method |
||
|
Diacerein |
Aceclofenac |
Diacerein |
Aceclofenac |
|
|
Linearity range |
2.5-15µg mL-1 |
5-30µg mL-1 |
2.5-15µg mL-1 |
5-30µg mL-1 |
|
Coefficient of correlation |
0.9998 |
0.9998 |
0.9999 |
0.9985 |
|
LOD |
0.12 |
0.58 |
0.18 |
0.52 |
|
LOQ |
0.36 |
1.17 |
0.54 |
1.57 |
|
Precision-Intraday*% RSD |
0.67 |
1.14 |
0.63 |
0.65 |
|
Precision-Interday*% RSD |
0.21 |
0.15 |
0.35 |
0.49 |
*-Average of six determinations, %RSD-Percentage Relative Standard Deviation, LOD-Limit of detection, LOQ- Limit of quantitation
Table 2: Results of recovery study of tablet formulation containing diacerein and aceclofenac by Vierodt’s Method and First Derivative method
|
Level of % Recovery(n=3) |
Vierodt’s Method |
First Derivative method |
||
|
Diacerein |
Aceclofenac |
Diacerein |
Aceclofenac |
|
|
50 |
101.28 |
102.37 |
99.25 |
98.24 |
|
100 |
99.98 |
107.48 |
96.49 |
98.85 |
|
150 |
100.34 |
100.68 |
96.82 |
100.32 |
|
Mean |
100.53 |
103.51 |
97.52 |
99.13 |
|
±SE |
0.28 |
0.057 |
0.85 |
0.50 |
±SE: Standard error
Precision:
Precision of the method was evaluated by using tablet powder equivalent to 100% of the label claim of diacerein and aceclofenac. Method repeatability was obtained by repeating assay of six replicates of single concentration of homogeneous sample, three times in a same day. Intermediate precision was assessed by assay of six replicates of single concentration of homogeneous sample of diacerein and aceclofenac on three consecutive days. Method precision was expressed as % R.S.D.
Recovery:
A recovery study was carried out by addition of known amount of standard drug in the preanalysed tablet formulation, in 50%, 100% and 150% of label claim. At each level of concentration three determinations were performed.
Limit of detection and limit of quantitation:
The detection limit and quantitation limit was computed for lower limit of detection and minimum quantity of analyte measured by proposed spectrophotometric methods.
RESULTS AND DISCUSSION:
The proposed Spectrophotometric methods were found to be more economic and simpler than earlier reported [9] method. The earlier reported methods [1-13], are developed using different instruments like LC-MS/MS, HPLC, narrowbore HPLC using column-switching, Spectrophotometric and spectrofluorimetric, reverse phase HPLC, capillary electrophoresis, in human plasma by HPLC, for estimation of diacerein and aceclofenac alone or in combination with other drugs, but we propose here two novel spectrophotometric methods for estimation of diacerein and aceclofenac in combined tablet dosage form. The proposed methods are rapid, no special sample preparation and extraction step involved for the estimations.
In Vierodt’s Method standard error was found to be less as compared to first derivative method. Correlation coefficient was found to be 0.9998 and 0.9999 for diacerein and 0.9998 and 0.9985 for aceclofenac by Vierodt’s method and by First Derivative method respectively (Table1). The limit of detection and quantitation were found 0.12 µg/ml and 0.36 µg/ml for diacerein and 0.58 µg/ml and 1.77 µg/ml for aceclofenac by Vierodt’s method and 0.18 µg/ml, 0.54 µg/ml for diacerein and 0.52 µg/ml, 1.57 µg/ml for aceclofenac respectively by First Derivative method (Table1). Method precision (% RSD) was found satisfactory in Vierodt’s method as compared to First Derivative method (Table1). Average percentage recovery was found to be 100.53 ± 0.28 for diacerein and 103.51± 0.057 for aceclofenac by Vierodt’s method and 97.52 ± 0.85 for diacerein and 99.13 ± 0.50 for aceclofenac by first derivative method (Table2).
CONCLUSION:
The proposed Spectrophotometric methods have proved to be convenient in procedure and economic as compared to Chromatographic methods. The developed Spectrophotometric methods can be used in routine estimation of diacerein and aceclofenac in pharmaceutical solid dosage forms.
ACKNOWLEDGEMENTS:
The authors gratefully acknowledge the Blue Cross Laboratories Pvt. Ltd., Nashik, India, for providing gift sample of diacerein and aceclofenac. The authors are also thankful to Principal V. M. Aurangabadkar and Management of M. G. V’s Pharmacy College, Panchavati, Nashik for providing necessary facilities and constant support for research work.
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Received on 15.09.2009 Modified on 08.12.2009
Accepted on 24.12.2009 © AJRC All right reserved
Asian J. Research Chem. 3(1): Jan.-Mar. 2010; Page 94-97