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paracetamol, aceclofenac and tizanidine from tablet dosage form. Methanol (95%) was used .... metamizol, paracetamol and caffeine in tablets,. Ana. Chim. Acta., 1998 ... aspirin and paracetamol by double divisor ratio spectra derivative and ...
International Journal of ChemTech Research CODEN( USA): IJCRGG ISSN : 0974-4290 Vol. 3, No.2, pp 963-966, April-June 2011

Validated Simultaneous Multicomponent Spectrophotometric determination of Paracetamol, Aceclofenac and Tizanidine in tablets Sampada Sinha*, Mithun Singh Rajput College of Pharmacy, IPS Academy, Rajendra Nagar, A.B. Road, Indore- 452012, India *Corres. author: [email protected] Phone no. +91- 9893409914 Abstract: Simple multicomponent spectrophotometric method has been developed for simultaneous estimation of paracetamol, aceclofenac and tizanidine from tablet dosage form. Methanol (95%) was used as solvent. The multicomponent mode of analysis involves the measurement of absorbances at three wavelengths 248 nm (λmax of paracetamol), 276 nm (λmax of aceclofenac) and 319 nm (λmax of tizanidine). The linearity for the method lies between 214 µg/ml for paracetamol, 5-40 µg/ml for aceclofenac and 5-25 µg/ml for tizanidine. The accuracy and precision of the method was determined and validated stastically. The method showed good reproducibility and recovery with % RSD less than 2. The method was found to be rapid, specific, precise and accurate and can be successfully applied for the routine analysis of paracetamol, aceclofenac and tizanidine in bulk and combined dosage form. Keywords: Paracetamol, Aceclofenac, Tizanidine, Multicomponent method.

Introduction Paracetamol and aceclofenac are NSAIDs whereas tizanidine is the drug of skeletal muscle relaxant category. Paracetamol is N-(4-hydroxyphenyl) acetamide. It is antipyretic and analgesic [1]. Aceclofenac is chemically known as 2-[2-[2-(2,6Dichlorophenyl)aminophenyl]acetyl]oxyacetic acid and is used in the treatment of osteoarthritis [1]. Tizanidine, 5-chloro-4-(2-imidazolin-2-ylamino)-2, 1, 3-benzothiadiazole is an α-2 adrenergic agonist and centrally active myotonolytic skeletal muscle relaxant with a chemical structure unrelated to other muscle relaxants [2]. Several spectrophotometric methods have been reported for estimation of these three drugs in various dosage forms with other drugs [3-7]. Since no spectrophotometric method is reported for simultaneous estimation of paracetamol, aceclofenac and tizanidine in combination, therefore in the present work, a successful attempt has been made to estimate these drugs simultaneously by simple UVspectrophotometric method.

Experimental Instrument, reagent and chemicals A Shimadzu UV-Visible recording spectrophotometer (Model-UV 1601) with 1 cm matched silica cells was used for spectrophotometric analysis. Paracetamol, aceclofenac and tizanidine bulk drugs were obtained as a gift sample from Alpa Laboratories Ltd., Indore, India, IPCA Laboratories Ltd., Ratlam, India and Unichem Laboratories Ltd., Roha, India respectively. Combined dose tablet formulation (Zerodol-MR) manufactured by IPCA Laboratories Ltd., India, was procured from local pharmacy, Indore, India. All other chemicals used were of analytical grade. Preparation of stock solution Ten milligram of paracetamol standard drug was accurately weighed and transferred to a 10 ml volumetric flask. To this, sufficient amount of 95% methanol was added and the flask was shaken to solubilize the drug. The volume was made up to the mark with methanol to obtain 1 mg/ml stock solution.

Sampada Sinha et al /Int.J. ChemTech Res.2011,3(2)

Similarly (1 mg/ml) stock solutions of aceclofenac and tizanidine were prepared. Multicomponent mode of analysis Five mixed standard solutions of paracetamol, aceclofenac and tizanidine were prepared in the ratio of 1:1:5 respectively having concentration in µg/ml of 2:2:10, 3:3:15, 4:4:20, 5:5:25 and 6:6:30 and analyzed in multicomponent mode at their respective λmax that is

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248, 276 and 319 nm respectively. The concentration of the individual drug was feed to the multicomponent mode of the instrument. The instrument collects and compiles the spectral data from mixed standards and the concentration of the various components of the formulation are directly recorded when the sample solution is scanned. The analysis was repeated five times. Multicomponent overlay spectra of paracetamol, aceclofenac and tizanidine are shown in figure 1.

Figure 1. Multicomponent overlay spectra of paracetamol, aceclofenac and tizanidine.

Table 1 :Recovery study data for tablet formulation Level of % Mean recovery % S.D % ACE PCM TIZA ACE PCM TIZA recovery 101.38 101.35 101.38 1.4 1.52 1.4 80 100.75 100.76 100.75 0.456 0.45 0.456 100 102.26 102.07 102.2 0.82 1.0 0.82 120 PCM: Paracetamol, ACE: Aceclofenac, TIZA: Tizanidine

% R.S.D ACE

PCM

TIZA

1.38 0.452 0.8

1.5 0.44 0.97

1.38 0.452 0.80

Table 2. Result of analysis of tablet formulation Parameters Linearity Accuracy Precision LOD LOQ

80 % 100 % 120 % Intraday Interday

Observation of multicomponent analysis PCM 2-14 µg/ml 101.35±1.52 100.76±0.45 102.07±1 99.6±0.35 99.7±0.31 0.05 µg/ml 0.153 µg/ml

ACE 5-40 µg/ml 101.38±1.4 100.75±0.45 102.26±0.82 99.4±0.76 100.23±0.471 0.114 µg/ml 0.34 µg/ml

100.02±1.4 100.62±1.97 Analysis of tablet PCM: Paracetamol, ACE: Aceclofenac, TIZA: Tizanidine

TIZA 5-25 µg/ml 101.38±1.4 100.75±0.456 102.2±0.82 99.6±0.35 99.3±0.31 0.104 µg/ml 0.31 µg/ml 100.64±1.9

Sampada Sinha et al /Int.J. ChemTech Res.2011,3(2)

Results and Discussion The developed method is a multicomponent analysis. It was observed during studies that there is significant interference between the analytes. Therefore area under curve method could not be developed. The derivative method was not developed because after derivatization of overlay spectra of paracetamol, aceclofenac and tizanidine, zero crossing point was not observed. Recovery study To check the accuracy of the developed method and to study the interference of formulation additive, analytical recovery experiment was carried out by standard addition method. Recovery study was performed by adding 80, 100, and 120% of the test concentration as per ICH guidelines [8-10]. The results and statistical validation of paracetamol, aceclofenac and tizanidine are reported in table 1. Intermediate precision (intra day and inter day precision) The intra and inter day precision was calculated by assay of the sample solution on the same day and on different days at different time interval respectively (Table 2). Limit of detection and limit of quantitation (LOD) and (LOQ) Limit of detection and limit of quantitation was calculated by the following formula

References 1. Mahaparale P.R., Sangshetti J.N. and Kuchekar B.S., Simultaneous estimation of aceclofenac and paracetamol in tablet dosage form, Ind. J. Pharm. Sci., 2007, 69,289-292. 2. Subramaniam G., Musmade P. and Agarwal S., Simultaneous estimation of tizanidine, diclofenac potassium and paracetamol in tablets, Ind. J. Pharm. Sci., 2004, 5,694-696.

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LOD = 3.3 x mean standard deviation/slope of curve LOQ = 10 x mean standard deviation/slope of curve Analysis of commercial tablet preparation Twenty tablets were weighed accurately, crushed and weight equivalent to 2 mg of tizanidine and 98 mg of pure drug was taken and then dissolved in 75 ml of methanol with frequent shaking for 30 min in a 100 ml volumetric flask. The final volume was made up to the mark with methanol. The sample solution was then filtered through Whatmann filter paper no. 4. This will produce solution containing 1000 µg/ml of tizanidine. Then 1 ml from the above solution was taken in 10 ml of volumetric flask and the final volume was made upto the mark with methanol. This will produce solution containing 100 µg/ml of tizanidine and corresponding concentration of paracetamol and aceclofenac. The results and statistical validation of paracetamol, aceclofenacqnd tizanidine are reported in table 2.

Conclusion In the research work done, a successful attempt for selective estimation of paracetamol, aceclofenac and tizanidine in three component tablet formulation by spectrophotometric method was made by experimentation based on literature survey. However, simultaneous estimation could not be achieved using simultaneous equation, area under curve and derivative method.

5. Dhake A.S., Sonaje D.B., Nikam P.T. and Talekar R.S., Simultaneous estimation of mefenamic acid and paracetamol by UV spectroscopy, Ind. J. Pharm. Sci., 2001, 1,55-54. 6. Erk N., Ozkan Y., Banoglu E., and Ozkan S.A., Simultaneous estimation of paracetamol and methocarbamol by ratio spectra derivative spectrophotometry and LC, J. Pharm. Biomed. Analysis, 2001, 24,469-475.

3. Dinc E. and Onur F., Application of a new spectrophotometric method for analysis of metamizol, paracetamol and caffeine in tablets, Ana. Chim. Acta., 1998, 359,93-106.

7. Nagulwar V., Dhurvey Y.R., Deshpande S. and Upadhye K., UV spectrophotometric simultaneous estimation of valdecoxib and paracetamol in tablet dosage form, Ind. J. Pharm. Sci., 2006, 68;639-641.

4. Dinc E., The spectrophotometric multicomponent analysis of ascorbic acid, aspirin and paracetamol by double divisor ratio spectra derivative and ratio spectra zero crossing methods, Talanta, 1999, 48,1145-1157.

8. International Conference on Harmonization (ICH), Q2A Text on Validation of Analytical Procedures: Definitions and Terminology, Vol. 60, US FDA Federal Register, 1995.

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9. International Conference on Harmonization (ICH), Q2B, Validation of Analytical Procedures: Definitions and Terminology, Vol.60, US FDA Federal Register, 1995.

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10. Sharma B.K., Instrumental methods of chemical analysis, Goel Publishing house, Meerut, 2005,C286-C311.