Assessment of Naproxen and Paracetamol in Mixed Tablet

Assessment of Naproxen and Paracetamol in Mixed Tablet METHOD DEVELOPMENT AND VALIDATION FOR SIMULTANEOUS ASSESSMENT OF NAPROXEN AND PARACETAMOL IN MIXED TABLET DOSAGE FORM BY RP-UPLC K.KANAKAPARVATHI*, Vijay Nagarjan, Santha Arcot and CH Hemanth Kumar. ABSTRACT An advancement design and corroboration for simultaneous assessment of Naproxen (NAP) and Paracetamol (PAR) in merged tablet dosage form by UPLC. The column used in determination was C18 thermo fisher (50cm x 4.6 mm x 3Â µm), mobile phase used in this method was 0.4% ammonium acetate buffer: methanol: acetonitrile (40:40:20), the retention time was about 1.9 minutes and 3 minutes for PAR and NAP of a total run time of 5 minutes, with flow rate of 0.2ml per minute respectively at a wavelength of 271nm, linearity of the method was linear over the range of 38.496 to 57.664ÃŽ ¼g/ml for Paracetamol and 64.096 to 95.968ÃŽ ¼g/ml of Naproxen respectively with a correlation of 0.999 for simultaneous assessment for PAR and NAP thus the method was fast, simple, elegant and less time consuming method Keywords: Naproxen, Paracetamol RP-UPLC, Method validation INTRODUCTION Naproxen is chemically 2-Naphthaleneacetic acid, 6-methoxy-ÃŽ ±-methyl-(s)-(+)-(s)-6 methoxy-ÃŽ ±-methyl-2-naphthaleneacetic acid as shown in (Figure 1). It is a non-steroidal anti-inflammatory drug commonly used for minimizing of moderate to severe torment, delirium, inflammation and stiffness. [6-11]. Paracetamol (PAR) is chemically N-(4-hydroxyphenyl) acetamide (Figure 2), It has analgesic and antipyretic activity for the therapy of subsidiary, non-inflammatory conditions of patient who were prone to gastric symptoms [12-14]. The merger of these two drugs are used in the remedy [11] of Musculoskeletal Disorder (Sprain/Strains) Trauma Fractures/injuries), Occupational affliction, Joint torment, Low Back laceration the literature review supports legion UPLC methods for the evaluation of NAP and PAR independently and in combination with other drugs but There was no UPLC method had been reported for the determination of NAP and PARA in merged dosage form So an experiment was taken to expand and corroborate a rapid RP-UPLC method [1-5] for the determination of NAP and PARA in mixed tablet dosage forms. Figure 1 NAPROXEN Figure 2 Paracetamol MATERIALS NAP and PAR was earned from Ideal analytical and research institution puducherry, India. All chemicals worn were analytical standard. The pharmaceutical tablet dosage form used in this study was NAPROSYN P with a label claim of NAP 300mg and PAR 500mg were purchased from local pharmacy. INSTRUMENTATION AND APPARATUS The uplc system used for advancement design and corroboration was thermo accela equipped with 1050 quaternary pump auto sampler and photodiode array detector. The detector output were recorded and processed using chrome quest software version 5.0 sonicator (PCI bath sonicator ) was used for degassing of mobile phase and sonication of the solutions prepared SOFTWARE: The statically calculation for the analysis was performed by using Microsoft excel 2010 software (Microsoft, USA) METHOD CORROBORATION: SYSTEM SUITABILITY: System suitability was determined by injecting the standard solution and observed the parameters like retention time, peak area, relative standard deviation, tailing factor, USP theoretical plates. LINEARITY For testing of linearity five different concentration of sample solution (80%, 90%, 100%, 110%, and 120%) was injected and checked over by plotting the graph as peak area verses concentration thus the data treated by linear regression analysis. ACCURACY Accuracy can be done by injecting the sample solution with known standard concentration and the amount of percentage recovery gives the accuracy of sample. PRECISION Precision can be evaluated by Interday and intraday, were the same sample solution has to be assayed for the same day and on different days at different time intervals ROBUSTNESS The determination of robustness can be done by changing the experimental condition deliberately. The condition may include of changing in mobile phase flow rate, pH and temperature, the percentage of RSD, tailing factor, resolution, were cross check with the original data. RESULT DISCUSSION: The method has validated according to the norms of international harmonization of conference (ICH) guidelines with regards of system suitability, linearity, accuracy, precision and robustness as follows SYSTEM SUITABILITY The system suitability tests were carried out to evaluate the resolution and reproducibility of the system for the analysis. The results of the system suitability test were summarized in Table No.1. Table 1: System suitability results S.No PARAMETERS PAR NAP 1 Retention Time 1.807 3.007 2 Peak area 410801 306340 3 Percentage area 57.28 42.72 4 Theoretical plates 2633 3306 5 Resolution 0.0000 0.85712 6 Tailing factor 1.754 1.696 Solution stability The solvents which had been used in the mobile phase were cost effective than the solvents used in the other UPLC methods which are reported in the literatures. Standard and samples solution stability was studied above 12 and 24 hours and found stable against the freshly prepared standard. Table2. Results of Solution stability Time (hrs) Percentage Assay Percentage difference in assay PAR NAP PAR NAP Initial 99.92 99.99 0.002 0.001 After 12 hrs 99.52 99.57 0.003 0.002 After 24 hrs 99.12 99.19 0.001 0.002 LINEARITY Linearity of the method was evaluated at 5 different concentration levels of 38.496 to 57.664ÃŽ ¼g/ml for Paracetamol and 64.096 to 95.968ÃŽ ¼g/ml of Naproxen respectively. Both the drugs were found to give linear detector response in the concentration under study with correlation coefficient of 0.997 and 0.999 for PAR and NAP respectively. Table3: Linearity study for NAP and PAR S.NO PARAMETERS PAR NAP 1 Linearity range 38.49 57.664ÃŽ ¼g/ml 64.09 -95.96ÃŽ ¼g/ml 2 Correlation coefficient (r2) 0.997 0.999 3 Slope 3769.8726 2867.1591 4 Intercept 1567.7362 0.1591 ACCURACY Accuracy of the method was determined by recovery test. The percentage recovery was found to be within the concentration of 100 to 115 as 100, 105, 110, and 115 (Table4). All results indicate that the method is highly accurate. Table: 4(a) accuracy data for PAR S.NO ACCURACY LEVEL STANDARD AREA SAMPLE AREA Mg/tab PERCENTAGE 1 100 404871 393726 499.83 99.97 2 105 404871 413927 525.48 105.1 3 110 404871 433143 549.87 109.97 4 115 404871 454077 576.46 115.29 Table 4(b) accuracy data for NAP S.NO ACCURACY LEVEL STANDARD AREA SAMPLE AREA Mg/tab PERCENTAGE 1 100 306460.4 303506 299.26 99.75 2 105 306460.4 319467 315.00 105.00 3 110 306460.4 334246 329.57 109.86 4 115 306460.4 350847 345.94 115.31 PRECISION This method was validated for its inter-day and intra-day precision. The results (table4) obtained were within the acceptable limit. Table 5: results for precision studies s.no Parameter(units) PAR NAP STANDARD AREA SAMPLE AREA PERCENTAGE STANDARD AREA SAMPLE AREA PERCENTAGE 1 Interday precision (1st day) (2nd day) (3rd day) 404871 404871 404871 401886 402568 403442 100.87 99.28 100.74 306460 306460 306460 307076 307209 309589 99.77 98.08 100.07 2 Intraday precision 1sthrs 2nd hrs 3rd hrs 404871 404871 404871 402645 401507 400271 100.17 100.65 100.49 306460 306460 306460 309957 307438 307946 99.82 99.76 100.07 3 Average 100.366 99.595 4 SD 0.584 0.75 5 RSD 0.582 0.758 ROBUSTNESS The robustness of the method was determined and the percentage RSD of the results was found to be less than 2.0%, which demonstrate that the developed method is robust. Table6. Results of Robustness parameter CHANGED PARAMETERS FLOW RATE WAVE LENGTH S.NO PARAMETERS 190 210 269 273 PAR NAP PAR NAP PAR NAP PAR NAP 1 Retention time 1.938 3.215 1.70 2.832 1.810 3.005 1.810 3.007 2 Area 462947 347334 406134 306784 432154 322852 426295 347442 3 % area 57.13 42.87 56.97 43.03 57.24 42.76 55.10 44.90 CONCLUSION: Thus, the above stated method for determination of PAR and NAP by UPLC method concludes as it can be quantified simultaneously by using of isocratic mobile phase of 0.4% ammonium acetate buffer: methanol: acetonitrile (40:40:20), by using of PDA detector at 271 nm. Thus the proposed method is simple, precise, accurate, rapid and sensitive, where it can be applied successfully for the assessment of PAR and NAP in combined pharmaceutical formulations. ACKNOWLEDGEMENT The authors are thankful to ideal analytical and research laboratory pondycherry, India for all the facilities provided to complete our work.