Research Articles

March-April 2017  |  Vol: 3  |  Issue: 2
UV Spectrophotometric Method Development and Validation for Quantitative Estimation of Diclofenac Sodium

Sagar Savale*, Hitendra Mahajan

 

*Department of Pharmaceutics, R. C. Patel institute of pharmaceutical education and research, Shirpur 425-405, dist. Dhule, Maharashtra, India

* Corresponding author,

Mr. Sagar Savale

Department of Pharmaceutics,

R. C. Patel Institute of Pharmaceutical

Education & Research, Shirpur, 425405,

dist. Dhule, Maharashtra, India.

Mobile No: +91 9960885333,

Email ID: avengersagar16@gmail.com

 

ABSTRACT

Aim: UV Spectrophotometric Method Development and Validation for quantitative estimation of Diclofenac Sodium. Objective:  U.V Spectrophotometric method have been widely employed for determination of analyte in a mixture. Our aim is to develop spectroscopic method for estimation of the diclofenac sodium in ternary mixture by using U.V spectrophotometry. Methodology: The method was validated as per ICH guidelines. The recovery studies confirmed the accuracy and precision of the method. Conclusion: It was successfully applied for the analysis of the drug in bulk and could be effectively used for the routine analysis. Key words: Diclofenac sodium, UV spectrophotometric method, Validation.

 

Introduction

       Diclofenac sodium belong to the family of non-steroidal anti-inflammatory drugs (NSAID) or cyclo-oxygenase (COX) inhibitors. It is an effective anti-inflammatory, analgesic and antipyretic agent. It is commonly used in the treatment of acute and chronic pain, rheumatoid and osteoarthritis. Chemically it is 2-(2, 6- dichlorophenyl) amino benzeneacetic acid 4-(3H 1, 2, dithiol-3-thione-5-yl) phenyl ester and is a low-molecular-weight drug (MWt: 318.13). The

present work describes the development of a simple, precise, accurate and reproducible

spectrophotometric method for the estimation of Diclofenac sodium in pharmaceutical dosage forms. The developed method was validated in accordance with ICH Guidelines and successfully employed for the assay Pharmaceutical preparation and dosage form (Kashiwame et al., 2011; Dangre et al., 2015; Sawale et al., 2015).

 

Material and Method

Material

        Diclofenac sodium supplied as a gift sample by Loba chem. Pvt. Ltd (Mumbai, India) used as working standard.

Instrumentation

       A double beam UV-VIS spectrophotometer (UV-1700, Shimadzu, Japan) connected to a computer loaded with spectra manager software UV Probe was used. The spectra were obtained with the instrumental parameters as follows: Wavelength range: 200–800 nm. All weights were taken on an electronic balance (Model Shimadzu AUX 120).

 

Preparation of standard stock solution

      According to European pharmacopoeia, 10 mg of Diclofenac sodium was dissolve in 100 ml of methanol (100 µg/mL). Out of this stock 0.2-1.2 ml was pipetted and diluted up to 10 ml by methanol (2-12 µg/mL) and examined between 200-400 nm. The maximum absorbance was determined using UV-Vis Specrophotometer (UV-1700, Shimadzu, Japan) to confirm the λmax of the drugs.

 

Validation of analytical method

       The analytical performance characteristics which may be tested during methods validation: % Recovery, Precision, Ruggedness and sensitivity (Han et al., 2003; Sharma et al., 2005; Aggarwal et al., 2006).

 

Results and Discussion

Method Development

       The solution of diclofenac sodium in methanol was found to exhibit maximum absorption at 276 nm after scanning on the UV-Vis spectrophotometer which was reported as λmax in the literature and the procured drug sample of diclofenac sodium complies with the reference spectra (Figure 1).

Figure 1. UV spectra of Diclofenac sodium

 

 

 Linearity study

      Accurately weighted diclofenac sodium (10 mg) was dissolved in 100 ml of methanol to obtain working standard of 100 μg/ml. Aliquots were pipetted from the stock solution of drug and were transferred to 10 ml volumetric flask, the final volume was adjusted with methanol so that concentration of 2-12 μg/ml could be made. Absorbance of the above solution were taken at 276 nm by using UV-Vis spectrophotometer (UV-1700, Shimadzu, Japan) against the blank solution prepared in the same manner without adding the drug. A graph of absorbance vs concentration was plotted (Figure 2) and R2 was found to be 0.9986.

 

Figure 2. Calibration curve of Diclofenac sodium

Validation of analytical method

Recovery

       Recovery study is performed by standard addition method by adding the known amount of diclofenac sodium (Working standard) at two different concentration levels i.e 80%, 100% of assay concentration and % recovery for all these drug were calculated. Result was reported in Table 1.

 

Table 1. Recovery study

Drug

Initial amount

(µg/ml)

Added Amount (µg/ml)

% Recovery

% RSD

(n = 3)

Diclofenac sodium

2

1.9

100.15

0.07

 

2

1.8

100.98

0.05

 

 

 

 

 

 

Precision

     Intra-day precision was determined by analysing, the two different concentrations 2 mg/ml, 3 mg/ml containing diclofenac sodium, for three times in the same day (n = 3) Table 2. Inter-day variability was assessed using above mentioned three concentrations analysed on three different days, over a period of one week (n = 3) Table 2.

 

 

 

Table 2. Presion study

 

 

Intra - Day

Inter - Day

Drug

Con. (µg/ml)

Mean ± SD

% RSD

Mean ± SD

% RSD

Diclofenac sodium

2

2.0 ± 0.0016

0.01

1.9 ± 0.0014

0.07

 

3

2.9 ± 0.0014

0.05

2.9 ± 0.0048

0.06

 

 

 

 

 

 

 

Ruggedness

      From stock solution, sample solution containing diclofenac sodium (2 µg/ml) was prepared and analyzed by two different analysts using similar operational and environmental conditions (Table 3) (n = 3).

 

 

Table 3. Ruggedness study

 

% Amount Found

% RSD

Drug

Analyst I

Analyst II

Analyst I

Analyst II

Diclofenac sodium

100.44

99.44

0.01

0.05

 

 

 

 

 

Sensitivity

       Sensitivity of the proposed method was estimated in terms of Limit of Detection (LOD) and Limit of Quantitation (LOQ) (Table 4).

 

Table 4. Sensitivity study

Drug

LOD

LOQ

Diclofenac sodium

0.19 ± 0.004

0.44 ± 0.011

 

 

 

 

Conclusion

     The proposed UV spectrophotometric method was found very simple, rapid and economical. The method is validated in compliance with ICH guidelines is suitable for estimation of diclofenac sodium with excellent recovery, precision and linearity.

 

Acknowledgements

       The authors are grateful to Hon. Principal, SES’s, R. C. Patel Institute of Pharmaceutical Education and Research, Dr. S. J. Surana and wish to acknowledge all those who are involved directly or indirectly for compilation of this article. It has been a great honour to work with such a professional.

 

Reference

Aggarwal BB, Shishodia S. 2006. Molecular targets of dietary agents for prevention and therapy of cancer. Biochemical Pharmacology, 71: 1397- 421.

Dangre P, Sawale V, Meshram S, Gunde M. 2015. Development and validation of RP-HPLC method for the Simultaneous Estimation of Eprosartan mesylate and chlorthalidone in Tablet Dosage Form. International Journal of PharmTech Research, 8(2): 163-168.

Han W, Liu T, Himo F, Toutchkine A, Bashford D, Hahn KM, Noodleman LA. 2003. A theoretical study of the UV/visible absorption and emission solvatochromic properties of solvent-sensitive dyes. Chemphyschem, 4: 1084.

Kashiwame Y, Watanabe M, Araki K, Kuwata S, Ikariya T. 2011. Synthesis, Structure, and Proton-Transfer Reactions of Brønsted Acidic Pyridylpyrazole Complexes of Ruthenium. Bulletin of the Chemical Society of Japan, 84(3): 251.

Sharma RA, Gescher AJ, Steward WP. 2005. Curcumin: The story so far. European Journal of Cancer, 41: 1955-68.

Sawale V, Dangre P, Dhabarde D. 2015. Development and validation of RP-HPLC method for the simulteneous estimation of olmesartan medoxomil and chlorthalidone in tablet dosage form. International Journal of Pharmacy and Pharmaceutical Sciences, 7(5): 266-269.

 

 

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