Development and Validation of A UV Spectroscopic Method to Estimate Etoricoxib in Bulk and Tablet Formulation
Garlapati Manideep1, Nazare Lobo John Shane1, Girish Pai2,
Muddukrishna Badamane Sathyanarayana1*
¹Department of Pharmaceutical Quality Assurance, Manipal College of Pharmaceutical Sciences (MCOPS), Manipal Academy of Higher Education, Manipal, Karnataka, India.
2Department of Pharmaceutics, Manipal College of Pharmaceutical Sciences (MCOPS), Manipal Academy of Higher Education, Manipal, Karnataka, India.
*Corresponding Author E-mail: krishna.mbs@manipal.edu.
ABSTRACT:
This present UV Spectrophotometric method developed using methanol as a solvent is simple, rapid, specific, precise and sensitive to estimation Etoricoxib in bulk in day to day analysis. The method was validated as per ICH Q2R1 guideline. Etoricoxib is the newest addition to the group of Non-Steroidal Anti-Inflammatory Drugs - highly selective COX-2 inhibitor. The solvent used in the entire method development and validation was methanol. The maximum wavelength of absorption was found to be 234nm. Beer’s law was obeyed in the concentration range of 1 to 11µg/mL with correlation coefficient of 0.9986. The method was precise with an RSD of0.42%the LOD and LOQ were found to be 0.09µg/ml and 0.2µg/ml respectively. The method was validated for precision, linearity, accuracy and robustness and all parameters were found to be satisfactory which proves that this method can be used for routine analysis of Etoricoxib.
KEYWORDS: Etoricoxib, spectroscopy, validation.
1. INTRODUCTION:
Etoricoxib is cox-2 inhibitor. It is chemically 5-chloro-6-methyl-3-[(4-methyl sulfonyl )phenyl]-2-3-bi pyridine[1]. Its chemical formula is C18H15C1N2O2S. It is indicated for the treatment of rheumatoid arthiritis, osteoarthritis, gout inflammation and chronic low back pain. It has 106 fold selectivity for cox-2 inhibitors over cox-1 inhibitors[2].Etoricoxib an off-white crystalline powder is relatively insoluble in water, and freely soluble in alkaline aqueous solutions.During formulation development Analysis is an important step, and it should include a reliable, simple and cost-effective method. With this objective, the present research work was undertaken to develop and validate simple UV spectroscopic method to estimate Etoricoxib in bulk and tablet formulation.
Earlier methods including thin layer chromatography (HPTLC)[5,6], high performance liquid chromatography (HPLC),[3,4] high performance liquid chromatography-mass spectrometry (LC–MS)[7], ultra performance liquid chromatography (UPLC)[9] capillary zone electrophoresis,[8] and a UV method for quantification of ETX in pharmaceutical dosage forms which is not very sensitive[1]. These reported methods being costly and time consuming cannot be used for day to day analysis. Therefore a simple and sensitive UV spectrophotometric method was developed and validated as per International Conference on Harmonization (ICH) guidelines.
Figure 1- Structure of Etoricoxib
2. OBJECTIVE:
Development and Validation of A UV Spectroscopic Method to Estimate Etoricoxib in Bulk and Tablet Formulation
3. MATERIALS AND METHODS:
3.1 Instrument and materials:
A UV-Visible Spectrophotometer (Shimadzu UV-1800) with 10 mm quartz cells were used for analysis. Methanol and Acetonitrile (analytical grade) were obtained from Finar chemicals Ltd Ahmedabad. Etoricoxib 60mg tablets IP were procured from Zydus healthcare.
3.2 Preparation of standard stock solutions:
10mg of standard drug of Etoricoxib was weighed and transferred in to a 10ml volumetric flask and the volume was made up with 10ml of methanol which is used as a solventto get a concentration of 1000µg/ml. From the above solution the stock solutions of 100µg/ml and 10µg/ml were prepared. Different aliquots were taken from the stock solution of 10µg/ml and the volume was made up with methanol to get concentrations from 1-11 µg/ml which is the linearity range of this method. The λmax was found out by taking a spectrum over a range of 200-400nm and was found to be 235nm.
Figure 2: spectrum of etoricoxib standard
3.3 Preparation of sample stock solutions:
Marketed tablet formulation of Etoricoxib was used for analysis. 20 tablets were weighed and their average weight was determined. The tablets were then crushed in a mortar and pestle to form a powder. Tablet powder Equivalent to 10mg of Etoricoxib was taken into a 10ml volumetric flask and the volume was made up with methanol. The solution was then filtered to remove undissolved excipients. From the above solution the stock solutions of 100µg/ml and 10µg/ml were prepared.
4. RESULTS AND DISCUSSION:
After method development the next step is to validate the method for parameters like accuracy, linearity, LOD, LOQ, precision and robustness. The percentage relative standard deviation was calculated for all parameters to check whether they are within the acceptance limit. The proposed method was validated as per ICH Q2R1 guideline.
4.1 Linearity:
The ability of assay values to be directly proportional to the concentration of analyte in the sample is called linearity. Linearity was performed by taking aliquots of five or six different concentrations from the stock solution. The final volume was made up to the mark with methanol which is used as a solvent. Etoricoxib followed beer lamberts law in the concentration range of 1-11µg/ml.The linearity curve was prepared by plotting the concentration versus absorbance. A series of etoricoxib standard solutions were prepared over a range of 1-11µg/ml. Regression (r2) of 0.998 was obtained (figure 2).
Figure 3: linearity curve of Etoricoxib
4.2 LOD and LOQ:
Detection Limit is the smallest drug quantity which can be detected under normal test conditions. Quantitation Limit is the lowest drug concentration that can be accurately and precisely determined. LOQ and LOD were calculated according to ICH Q2R1 guideline. LOQ and LOD were determined Based on the Standard Deviation of the Response and the Slope. For proposed method LOQ and LOD was found to be 0.26µg/mL and 0.09µg/mL.
Table 1: LOD AND LOQ results of Etoricoxib
|
Parameters |
Obtained Values |
|
LOD |
0.09µg/mL |
|
LOQ |
0.26µg/mL |
4.3 Precision:
It is a measure of repeatability and reproducibility under unchanged conditions. The repeatability and reproducibility was determined by taking six aliquots of same concentration (5µg/mL) and absorbance was measured individually. The repeatability precision of the present method was obtained with a %R.S.D of 0.42. The reproducibility precision (inter-day) of the present method was obtained with a %R.S.D of 0.93. The results of the present method show that the method is precise and reproducible.
Table 2: precision results of Etoricoxib
|
Parameter |
%RSD |
Acceptance Criteria |
|
Repeatability |
0.42 |
NMT 1% |
|
Reproducibility |
0.93 |
NMT 2% |
4.4 Accuracy:
The accuracy was performed by recovery studies. Standard samples were used to spike a predetermined amount of sample drug from which the recovery was calculated. Accuracy was determined at 3 levels of 120, 100 and 80% of standard concentration. Four solutions were prepared containing respectively 100% sample solution (tablet). Into three of the above solutions 80%, 100% and 120% of the standard was spiked. The % recovery at the three levels were calculated and were found to be within the range according to ICH guidelines.
Table 3: accuracy results of Etoricoxib
|
Sample |
Absorbance |
%Recovery |
|
|
Test (5µg/ml) |
0.3242 |
||
|
Standard 80% |
0.2377 |
80% |
100.58% |
|
Standard100% |
0.3126 |
||
|
Standard120% |
0.4166 |
100% |
100.98% |
|
Mixture 80% |
0.5638 |
||
|
Mixture 100% |
0.6399 |
120% |
100.95% |
|
Mixture120% |
0.7440 |
||
4.5 Robustness:
It is defined as the capacity of the method to remain unaffected by small variation in analytical parameters. It is performed by taking six aliquots of same concentration (5µg/mL) andmeasuring the absorbance when wavelength was changed by ±1nm and by using a different solvent to prepare the solutions. The solvent used here is Acetonitrile. The %RSD of the results was calculated and it was found to be within the limit.
Table 4a: robustness results of Etoricoxib (by changing wavelength)
|
Parameter changed |
Observation |
Acceptance criteria |
|
|
234 nm |
236 nm |
||
|
Wavelength ±1nm |
0.4064 |
0.4536 |
NMT 2% |
|
0.4180 |
0.4560 |
||
|
0.4182 |
0.4429 |
||
|
0.4187 |
0.4523 |
||
|
0.4183 |
0.4540 |
||
|
0.4187 |
0.4576 |
||
|
0.4064 |
0.4536 |
||
|
% RSD |
1.17% |
1.14% |
|
Table 4b: robustness results of Etoricoxib (by changing solvent)
|
Parameter changed |
observation |
Acceptance criteria |
|
Solvent |
0.4077 |
NMT 2% |
|
0.4112 |
||
|
0.4119 |
||
|
0.4088 |
||
|
0.4123 |
||
|
0.4178 |
||
|
% RSD |
0.85% |
4.6 Assay:
Etoricoxib containing tablets were subjected to the analysis by the proposed method. The assay of the Etoricoxib tablets was 98.9 ± 0.49%(n=3). This results clearly indicate that the present method can be used for the routine quality control(QC) of Etoricoxib tablets without interference from the excipients and degradation products.
Table 5: assay results of Etoricoxib marketed formulation
|
Formulation |
Label Claim |
% Assay ±SD (n=3) |
|
Marketed Tablet |
Each tablet contains 60mg of Etoricoxib |
98.9 ± 0.49 |
5. CONCLUSION:
From the above experiment and results, it can be concluded that the newly developed and validated method for estimation of Etoricoxib was precise, accurate, sensitive and economical. Low inter-day and intra-day % RSD coupled with excellent recoveries were achieved which shows that this method can be applicable for routine lab analysis.
6. ACKNOWLEDGEMENT:
The author Thankful to Manipal Academy of Higher Educationand Department of Science and Technology, Government of India (for instrumentation support) under FIST scheme.
7. REFERENCES:
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Received on 22.07.2017 Modified on 18.09.2017
Accepted on 10.10.2017 © RJPT All right reserved
Research J. Pharm. and Tech 2018; 11(2):758-760.
DOI: 10.5958/0974-360X.2018.00142.7