UV- Spectrophotometric Method Development and Validation for Estimation of Tizanidine and Aceclofenac in Tablet Formulation

Jumle R. S.*, Mundhey A.S., Wate S.P., Dangare S.S., Ramteke U.D.

Department of Pharmaceutical Chemistry, Sharad Pawar College of Pharmacy Nagpur, India

*Corresponding Author E-mail: rjumle2010@gmail.com

ABSTRACT:

The present study deals with UV spectrophotometric method development and validation for estimation of Tizanidine and Aceclofenac tablet dosage form by Viedort’s method and first order UV derivative spectrophotometry. The Vierodt's method involves measurement of absorbance at λmax of Tizaridine and Aceclofenac at 282 nm respectively. The linearity of Tizanidine and Aceclofenac was found to be in the range of 1-10 µg/ml respectively. The % recovery of Tizanidine and Aceclofenac was found out to be 99.2% and 99.69%s respectively. First order CV derivative spectrophotonmetry (D1 method). the zero crossing method was chosen as Tizanidine could he easily analyzed without any interference from Aceclofenac and vice-versa. Tizanidine was determined by measurement of its D1 amplitude at the zero crossing point of Aceclofenac at (270nm), While Aceclofenac was determined by measurement of its D1 amplitude at zero crossing point of Tizanidine at (318 nm) The proposed method was validated as per ICH guidelines.

INTRODUCTION:

Tizanidine HCL [TIZ] chemically is chloro-N-imidazolin-2-y1)2,1,3-benzothiadiazol 4yl-amine. Tizanidine is a short acting drug for the management of spasticity. It is an monist at a 2- adrenergic receptor sites and presumably reduces spasticity by increasing presynaptic inhibition of motor neurons. Aceclofenac [ACE] chemically is 2-[2-(2,6-Dichlorophenyl) amino pheny] [ACE] chemically acid which is used as an effective NSAID having pronounced analgesic, antipyretic antiinflammatory property. It is belonging to developed NSAIDS of arylacetic acid type and structurally related to diclofenac. Aceclofenac-tizaniding combination is more effective than acrclofenac alone and had a favourable safety profile in the treatment of acute low back pain and for rheumatic disorders. Aceclofenac is reported for spectrophotometric, RP-HPLC art simultaneous estimation with other combinations^[1,3,5,10]Similarly, Tizanidine also reporter in combination with other drugs^[2,4,6,7,8]. Since no spectrophotometric method is reported for simultaneous estimation of Aceclofenac and Tizanidine in combination using methanol and distilled water (90:10).

The present paper describes a simple, accurate and precise method for simultaneous estimation of Aceclofenac and Tizanidine in combined tablet dosage form. TIZ and ACE is official in Indian Pharmacopoeia 2007 respectively^[10]Therefore, the present work, successful attempt has been made to estimate boil these drugs simultaneously by two simple UV spectrophotometric methods (Viedort’s) method and First order derivative method).

EXPERIMENTAL:

Solubility Studies:

Tizanidine is soluble in water but Aceclofenac is insoluble in water, hence various solvent systems were screened for simultaneous determination and methanol and Distilled water (90:10) was selected as solvent system

Instrumentation:

The instrument used was Shimadzu double beam UN/Vis spectrophotometer model V- 1700. Weighing was done on electronic balance (Shimadzu).

Table 1: Validation as per ICH guidelines

Parameters	Method I		Method II
Parameters	TIZ (318nm)	ACE (282nm)	TIZ (318nm)	ACE (270nm)
Linearity range (µgm)	1-10 µg/ml	2-20 µg/ml	1-10 µg/ml	2-20 µg/ml
Correlation coefficient (r²)	0.999	0.998	0.996	0.997
Interday	0.30	0.07	0.43	0.09
intraday	0.61	0.05	0.74	0.10
Slope	0.0344	0.1052	0.0317	0.3563
Intercept	0.0017	0.0003	0.0016	0.0004

Table 2: Result of recovery studies

% Mean recovery		Standard deviation		% RSD
TIZ	ACE	TIZ	ACE	TIZ	ACE
98.66	98.83	1.01	0.9	0.216	0.227
99.72	99.60	1.02	1.10	0.255	0.286
100.72	101.28	1.12	1.5	0.320	0.406

Materials:

TIZ HCL drug sample "as kindly supplied by Intra Lab, (Mumbai) and ACE drug sample \vas supplied by Zim Laboratories (Nagpur, India) and were used without any further purification. AR grad methanol was purchased from Merck Chemicals, India. Assay was carried out on Acenet-TZ tablet dosage form labeled to contain 2 mg of TIZ and 100 mg of ACE.

Preparation of Standard Stock Solutions:

Standard stock solutions of TZK and ACE were prepared separately by dissolving 100 mg of each drug in 10ml of solvent to get standard stock solution of 1000 µg/ml respectively by sonicating for 15 min and I ml was pipette out and further volume was made up to 10 ml with to obtain concentration of 100 µg/ml. Further dilutions were made in solvent buffer from stock solution to get concentrations of 1-10 µg/ml of TIZ and 2-20 µg/ml of ACE. The standard solutions of both TIZ, ACE were scanned in the range of 400-200 nm against solvent and spectra was recorded. λmax of TIZ and ACE was found at 318nm and 282 nm respectively.

Procedure:

From the stock solution of 100 µg/ml working standard solutions of drug were prepared by appropriate dilutions were prepared in solvent scanned in entire UV range to determine λmax, TIZ has λmax of 318 nm while ACE has λmax of 282nm respectively. Standard stock solution were prepared having concentration of 1-10 µg/ml of TIZ and 2-20 µg/ml, of ACE. The absorbance of these standard stock solution were measured at 318 nm and 282 and calibration curve were plotted.

The zero order spectra were recorded over 200-400 range for TIZ and ACE. As shown (Fig.1) the zero order spectra of pure drugs were found to be non overlapping. making simultaneous determination difficult. In contrast, the first derivative spectra of TIZ and ACE showed zero crossing points (Fig.2). The shape of the first derivative spectra is adequate for determining TIZ in the presence of ACE and vice versa. TIZ was determined by measurement of its D_Iamplitude at the zero crossing point of ACE at (270nm), While ACE was determined by measurement of its D₁at Zero crossing point of TIZ at (318 nm).

Procedure for the Analysis

Formulation:

Ten tablets containing label claim of 2 mg of TIZ and 100 mg of ACE were weighed and finely powdered. Weight of the powder equivalent to 0.0225 mg table was accurately weighed, transferred into a 100 ml flask, dissolved in solvent and this solution was sonicated for about 20 minutes filtered to separate any insoluble matter and volume was made up to 100 ml with solvent.

Fig. 1:- Overlain spectra of TIZ and ACE.

The clear solution obtained was diluted to get appropriate concentration in linearity ranges and absorbencies were measured.

Recovery Studies:

To study the accuracy of the proposed method, recovery studies were carried out at three different levels 80%, 100% and 120% by addition of known amount of TIZ and ACE to a knows concentration of the commercial tablet.

Fig. 2:- Overlain first derivative spectra of TIZ and ACE.

RESULTS AND DISCUSSION:

The present work provides an accurate rapid, sensitive method for the simultaneous analysis of TIZ and ACE in bulk -and tablet formulation. Linear relationships between D₁, amplitude and drug concentration were obtained over the range of at 1-10 and 2-20 µg/ml for TIZ and ACE respectively and also by simultaneous estimation respectively. The correlation coefficient, slope and intercept obtained for each drug is as shown in Table-1. The proposed method was also successfully applied to a pharmaceutical formulation. The % assay was

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Received on 12.07.2012 Accepted on 09.09.2012

Asian J. Pharm. Ana. 2(4): Oct. - Dec. 2012; Page 101-103