A VALIDATED STABILITY INDICATING RP-HPLC METHOD FOR ESTIMATION OF TOLFENAMIC ACID IN PRESENCE OF ITS PHARMACOPOEIAL IMPURITIES
DOI:
https://doi.org/10.22159/ijap.2019v11i5.34606Keywords:
Tolfenamic acid, Related impurities, RP-HPLC, ICH Q2 (R1) guidelines, Stability indicatingAbstract
Objective: The proposed research work was conducted to develop a single reverse-phase high-performance chromatography (RP-HPLC) method capable of separating two Pharmacopoeial related impurities as well as degradation product of Tolfenamic acid (TA). The drug was subjected to various stress conditions recommended under ICH Q1A (R2) guidelines.
Methods: The desired separation of two Pharmacopoeial impurities and one degradant generated under oxidative stress was carried out using Sunfire ODS C-18 (250 x 4.6 mm, 5 µm) column maintained at 40 °C. Isocratic elution was carried out using acetonitrile and ammonium dihydrogen orthophosphate buffer (10 mmol, pH 2.5) in the ratio of 80:20 v/v. The detection was carried out at 205 nm using flow rate of 1 ml/min. The developed method was validated as per ICH Q2 (R1) guidelines for specificity, linearity, accuracy, precision, Limit of detection (LOD), Limit of Quantification (LOQ) and robustness.
Results: Linearity response of TA was found at a concentration range of 10-100µg/ml, with a correlation coefficient of 0.9987. The Pharmacopoeial impurity A and impurity B showed linearity results at concentration of 0.1-1µg/ml, with correlation coefficient of 0.9984 for Impurity A and 0.9989 for Impurity B. The % recovery during accuracy studies for TA and the two impurities were within the acceptance range of 95-105%. LOD and LOQ for TA were found to be 4.561µg/ml and 133.771µg/ml respectively. For impurity A, LOD and LOQ were found to be 0.035 µg/ml and 0.106 µg/ml and for Impurity B, LOD and LOQ were 0.042 µg/ml and 0.128 µg/ml. With slight variation of organic phase in mobile phase and flow rate the method exhibited good robustness. Under forced degradation studies the drug was found stable under hydrolytic, photolytic and thermal stress conditions, but was found susceptible for degradation under oxidative stress with appearance of a degradant peak. From on the RRT values of Pharmacopoeial impurities and the formed degradant it was inferred that the developed method is selective for the drug in the presence of impurities or degradants.
Conclusion: The developed stability-indicating method is found to be simple, rapid, accurate, precise and robust as compared to other proposed methods while determining TA in presence of its Pharmacopoeial impurities and degradation products. Hence the developed method can be used for analysis of stability samples of TA in presence of its related impurities.
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References
The Merck Index. In: Budawari S. editor. 13th ed. Whitehouse Station, NJ: Merck and Co Inc; 2001.
British Pharmacopoeia. The department of Health. London. Stationary Office; 2009. p. 2244-5.
Martindale. The complete drug reference. In: Sweetman SC. editors. 34th ed. London: The Pharmaceutical Press; 2007.
Niopas I, Georgarakis M. Determination of tolfenamic acid in human plasma by HPLC. J Liq Chromatogr 1995;18:2675-82.
Colon J, Basha MR, Madero Visbal R, Kundari S, Baker CH, Herrera LJ, et al. Tolfenamic acid decreases c-Met expression through Sp proteins degradation and inhibits lung cancer cells growth and tumor formation in orthotopic mice. Invest New Drugs 2011;29:41-51.
Eslin D, Sankpal UT, Lee C, Sutphin RM, Maliakal P, Currier E, et al. Tolfenamic acid inhibits neuroblastoma cell proliferation and induces apoptosis: a novel therapeutic agent for neuroblastoma. Mol Carcinog 2013;52:377–86.
Kim JH, Jung JY, Shim JH, Kim J, Choi KH, Shin JA, et al. Apoptotic effect of tolfenamic acid in KB human oral cancer cells: possible involvement of the p38 MAPK pathway. J Clin Biochem Nutr 2010;47:74–80.
Liu X, Abdelrahim M, Abudayyeh A, Lei P, Safe S. The nonsteroidal anti-inflammatory drug tolfenamic acid inhibits BT474 and SKBR3 breast cancer cell and tumor growth by repressing erbB2 expression. Mol Cancer Ther 2009;8:1207–17.
Babu C, Devanna N, Suresh R. Validated gradient stability indicating RP-HPLC method for the simultaneous quantification of 11 related substances in the combined dosage forms of lamivudine and tenofovir disopeoxil fumarate. Int J Appl Pharm 2017;9:61-8.
Shim JH, Shin JA, Jung JY, Choi KH, Choi ES, Cho NP. Chemopreventive effect of tolfenamic acid on KB human cervical cancer cells and tumor xenograft by downregulating specificity protein 1. Eur J Cancer Prev 2011;20:102–11.
Tsiliou S, Kefala LA, Perdih F, Turel I, Kessissoglou DP, Psomas G. Cobalt (II) complexes with non-steroidal anti-inflammatory drug tolfenamic acid: Structure and biological evaluation. Eur J Med Chem 2012;48:132–42.
Ioannou PC, Rusakova NV, Andrikopoulou DA, Glynou KM, Tzompanaki GM. Spectrofluorimetric determination of anthranilic acid derivatives based on terbium sensitized fluorescence. Analyst 1998;123:2839–43.
Papadoyannis I, Georgarakis M, Samanidou V, Zotou A. Rapid assay for the determination of tolfenamic acid in pharmaceutical preparations and biological fluids by high-performance liquid chromatography. J Liq Chromatogr Relat Technol 1991;14:2951–67.
Gallo P, Fabbrocino S, Dowling G, Salini M, Fiori M, Perretta G, et al. Confirmatory analysis of non-steroidal anti-inflammatory drugs in bovine milk by high-performance liquid chromatography with fluorescence detection. J Chromatogr A 2010;1217:2832–9.
Ibrahim H, Boyer A, Bouajila J, Couderc F, Nepveu F. Determination of non-steroidal anti-inflammatory drugs in pharmaceuticals and human serum by dual-mode gradient HPLC and fluorescence detection. J Chromatogr B 2007;857:59–6.
Gonzalez Barreiro C, Lores M, Casais MC, Cela R. Simultaneous determination of neutral and acidic pharmaceuticals in wastewater by high-performance liquid chromatography-post-column photochemically induced fluorimetry. J Chromatogr A 2003;993:29–37.
Mikami E, Goto T, Ohno T, Matsumoto H, Inagaki K, Ishihara H, et al. Simultaneous analysis of anthranilic acid derivatives in pharmaceuticals and human urine by high-performance liquid chromatography with isocratic elution. J. Chromatogr B: Biomed Sci Appl 2000;744:81–9.
Rozou S, Antoniadou Vyza E. An improved HPLC method overcoming Beer’s law deviations arising from supramolecular interactions in tolfenamic acid and cyclodextrins complexes. J Pharm Biomed Anal 1998;18:899–905.
Niopas I, Georgarakis M. Determination of tolfenamic acid in human plasma by HPLC. J Liq Chromatogr Relat Technol 1995;18:2675–82.
Papadoyannis IN, Zotou AC, Samanidou VF. Simultaneous reversed-phase gradient-HPLC analysis of anthranilic acid derivatives in anti-inflammatory drugs and samples of biological interest. Liq Chromatogr Relat Technol 1992;15:1923–45.
Shinozuka T, Takei S, Kuroda N, Kurihara K, Yanagida J. Micro determination of anthranilic acid derivatives of anti-inflammatory drugs by high-performance liquid chromatography and its application to forensic chemistry. Eisei Kagaku 1991;37:461-6.
Abdelwahab NS, Ali NW, Zaki MM, Abdelkawy M. Validated chromatographic methods for simultaneous determination of tolfenamic acid and its major impurities. J Chromatogr Sci 2015;53:481-91.
International Conference on Harmonization (ICH), Stability testing of new drug substances and products, Q1A (R2); 2003.
International Conference on Harmonization (ICH) Harmonized Tripartite Guidelines, Validation of Analytical Procedures, Text, and Methodology, Q2 (R1), Parent Guidelines on Methodology Dated; 1996. p. 6.
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