Development and Validation of Diclofenac sodium in tablets using Simple UV Spectrophotometric method

 

Ragini V. Mane, Kritika Patel, Gude Sai Sushmitha, Dr. S G Vasantharaju*

Dept. of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences,

Manipal Academy of Higher Education, Manipal-576104, Karnataka, India

 

ABSTRACT:

An effective and simple UV-Spectrophotometric method was developed and validated for the quantification of Diclofenac sodium in tablets. The maximum absorption was found to be 282nm by using methanol as the solvent. The linearity range was 6-21 µg/ml and the correlation coefficient (r²) was found to be 0.9985. The method was accurate, precise, and robust with %RSD below 2%. The LOD and LOQ was found to be 0.0885μg/Ml and 0.250μg/mL respectively. Sandel’s sensitivity was found to be 0.000002 μg/mL. Assay and accuracy of the tablets were about of the acceptance criteria range as per the ICH guidelines.

 

 

 

INTRODUCTION:

Diclofenac sodium belongs to the class of dichlorobenzenes-organic compounds. It is a non-steroidal anti-inflammatory drug (NSAID) containing actions like antipyretic and analgesic. It chiefly exists in the form of sodium salt and as potassium salt. It is official in various pharmacopoeias such as United States Pharmacopoeia(USP), British Pharmacopoeia(BP) and Indian Pharmacopoeia (IP)¹

 

Diclofenac is mainly used in the treatment of pain, osteoarthritis, rheumatoid arthritis, dysmenorrhea, ocular inflammation, actinic keratosis and ankylosing spondylitis. The mechanism of action includes inhibition of leukocyte migration and the enzyme cyclooxygenase (COX-1 and COX-2), which leads to the peripheral inhibition of prostaglandin synthesis which is responsible for its analgesic effects. It acts on hypothalamus, resulting in dilationin peripheral region, leading to increase of cutaneous blood flow and subsequently leads to heat dissipation, which is responsible for its antipyretic effect².

 

Several methods are available in the literature for the diclofenac sodium and its combination with several other drugs. Most of all the diclofenac sodium analytical methods reported either of its API or formulation form for quantitative determination are based on Colorimetry, UV/Vis spectrometry, Reverse Phase High Performance Liquid Chromatography (RP-HPLC),HPTLC^3-7,9-16. These methods are time consuming and expensive.¹⁷ No UV Spectrometric is reported for the analysis of diclofenac sodium in API and tablets so far. Therefore, a simple and sensitive UV-Spectrophotometric method was developed and validated as per the International Conference on Harmonization (ICH) guidelines.

 

 

Figure 1 Chemical structure of Diclofenac sodium

 

 

 

MATERIAL AND METHODS:

Chemicals and reagents:

Diclofenac sodium was purchased from Yellow Pharma Mumbai. Commercial pharmaceutical formulation (tablet) containing 50 mg of diclofenac sodium was procuredfrom local market. Methanol and water were used of analytical grade of Qualigens Fine Chemicals make. A 0.45 µm pore size nylon filter of Pall life Sciences, Mumbai, India was used. Remaining chemicals used and agents used were analytical grade.

 

Instrumentation:

The entire proposed work was performed on UV-1800 series UV-visible spectrophotometer of Shimadzu make. It is with of 1 cm quartz cell and double beam and double detector configuration. The entire weighing was performed on Sartorius CP 225 D electronic balance. A Serve well clean ultrasonicator(India)was used for solubility purpose.

 

Selection of Solvent:

Based on solubility study performed, methanol was decided as the solvent (diluent) for dissolving diclofenac sodium.

 

Preparation of Standard Stock Solutions of Diclofenac sodium:

50mg quantity was accurately weighed and taken into a 50mL volumetric flask and into it 20mL of methanol was added and sonicated about 10min to dissolve the weighed drug. The volumetric flask was filled upto the mark with methanol and followed concentration would be 1mg/mL.

 

Diclofenac sodium-Working Standard Solution:

10 µg/ml of Diclofenac solution was prepared using methanol as a diluent.

 

 

Fig.2: Spectra of Diclofenac sodium

 

Determination of λ Max of Diclofenac sodium:

10 µg/ml of Diclofenac solution was scanned between 200 nm to 400 nm. The absorption maximum (λ max) was found to be at 282 nm (Figure2).282nm λ _maxwas considered for the whole study.

 

Linearity Study for Diclofenac sodium:

The necessary volume of the above working standard solution of Diclofenac sodium was taken into six individual volumetric flasks of 10mL. The final volume was filled up to the mark using methanol to obtain required concentrations (6-21 µg/mL). Absorbencies of these solutions were measured at 282 nm, (Table1) Calibration curve was constructed against its absorbance values in the Y-axis and concentration values in its X-axis as shown in (Fig. 3).

 

Table 1: Regression and Optical characteristics

Parameters	Value For Diclo
Beer’s law limit (µg/mL)	6-21
Correlation Coefficient (r)	0.9985
Regression equation
Slope	0.033
Intercept	0.061

 

 

 

Fig. 3: Calibration Curve of Diclofenac sodium at 282 nm Wavelength

Applications of the drug to estimate in tablets by using the proposed method:

Twenty ‘Vovaran-D’ Tablets holding 50 mg of Diclofenac sodium were weighed and ground to fine powder. The resulting tablet powder equivalent to 10mg of Diclofenac sodium was transferred into a 100ml volumetric flask, dissolved in methanol and sonicated for 10 minutes. The solution was filtered and 1.5ml was pipette out into a 10 ml volumetric flask and made up to the volume with methanol to obtain 15µg/ml solution. Absorbance of the resulting solution was measured at 282nm. From this, the percentage recovery was calculated which was found to be 103.53%. Therefore, the tablets passed the test as the acceptance criteria for diclofenac tablets is not less than 90% and not more than 110% as per IP, 2014.The results are tabulated in Table 2.

 

Table 2 Contentof Diclofenac sodium in Tablets.

Analyte	Label claim	% Label claim estimated (Mean ±S.D.)*	% R.S.D.
Diclofenac sodium	50 mg/tab	103.53 ± 0.676225	0.869922

*Average of 5readings; S.D. =Standard Deviation

 

Validation of Proposed Method:

According to ICH Q2R1 guidelines the method was validated⁸

 

Accuracy [Recovery Study]:

The proposed method was said to be accurate on the basis of recovery studies. The known amount of standard solutions were spiked into tablet powder(sample) solutions to obtained ultimate concentrations of 80%, 100% and 120% and analysed using proposed method. The absorbance values are recorded and the % recoveries were calculated.

 

% Recovery = [X - Y/ Z] X 100

 

Where,

X =The amount of standard and sample present in total.

Y = Amount of drug found as such

Z = Amount of core drug added.

 

The results are tabulated in (Table 3).

 

Table 3: Recovery Study.

S.D. =Standard Deviation

 

Precision:

Precision was determined as intra-day which in within the day and inter-day which is between days variations. Intra-day precision was determined by analysing Diclofenac sodium for three times on the same day at its 100% concentration level. Inter-day precision was determined by preparing the same concentration of solutions and analysed on three different days over a week. The obtained results are tabulated in (Table 4).

 

Table 4: The results of Precision

Precision	Diclofenac sodium	% R.S.D.
Interday, n = 3	99.05 ±0.2216	±0.298
Intraday, n = 3	99.01 ±0.1520	±0.1591

RSD = Relative standard deviation

 

Robustness:

It was performed by taking six solutions of the same concentration (10µg/ml) and the absorbance was measured when the wavelength was changed ± 2nm and by using a different solvent (water) to prepare the solutions. The %RSD are then calculated and was found to be within the limit. The obtained results are reported in (Table 5).

 

Table 5: Robustness Study (Wave length and solvent changed).

Parameter changed	observation
Wavelength±2nm	280nm	282nm	284nm
	0.3587	0.3590	0.3547
	0.3600	0.3608	0.3568
	0.3627	0.3635	0.3591
	0.3609	0.3615	0.3573
	0.3652	0.3662	0.3627
	0.3627	0.3634	0.3592
% RSD	0.75	0.825	0.90

 

Parameterchanged	Observation
Solvent (water)	0.2449 0.2466 0.2554 0.2485 0.2467 0.2502
RSD %	1.98

 

Limiting values:

Limit of detection of an individual analytical method is the lowest amount of detectable analyte in the sample. The quantification limit of an individual analytical method is the lowest amount of analyte in a sample which can be quantitatively determined with suitable precision and accuracy.

 

It is determined based on slope and standard deviation of the response

 

Table 5: LOD and LOQ values

Drug	LOD (µg/ml)	LOQ (µg/ml)
Diclofenac sodium	0.0885	0.250

 

ACKNOWLEDGEMENT:

The authors are grateful to the authorities of Manipal College of Pharmaceutical Sciences andManipal Academy of Higher Education, Manipal for the facilities

 

CONFLICT OF INTEREST:

The authors here by declare there is no conflict of interest.

 

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Accepted on 14.09.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(2):611-614.