Back
 AJAC  Vol.6 No.6 , May 2015
Development and Validation of Stability Indicating RP-HPLC Method on Core Shell Column for Determination of Degradation and Process Related Impurities of Apixaban—An Anticoagulant Drug
Abstract: A rapid, specific, sensitive, and precise reverse-phase HPLC method for the quantitative determination of process related and degradation impurities of Apixaban, an anticoagulant drug is described. The developed RP-HPLC method was successfully applied to the analysis of both Apixaban drug substance and drug product. The chromatographic separation was achieved on a Sigma-Aldrich’s Ascentis Express® C18 (4.6 mm × 100 mm, 2.7 μ) HPLC column with a runtime of 40 min. Mobile phase-A and mobile phase-B were phosphate buffer and acetonitrile respectively. The column oven temperature was set at 35°C and photodiode array detector was set at 225 nm. Nine process related impurities (Imp-1 to Imp-9) have been detected in test sample of Apixaban by using newly developed RP-HPLC method. Forced degradation study was carried out under acidic, alkaline, oxidative, photolytic and thermal conditions to demonstrate the stability-indicating nature of the developed RP-HPLC method. The developed method was validated as per ICH guideline and found to be specific, precise, sensitive and robust.
Cite this paper: Landge, S. , Jadhav, S. , Dahale, S. , Solanki, P. , Bembalkar, S. and Mathad, V. (2015) Development and Validation of Stability Indicating RP-HPLC Method on Core Shell Column for Determination of Degradation and Process Related Impurities of Apixaban—An Anticoagulant Drug. American Journal of Analytical Chemistry, 6, 539-550. doi: 10.4236/ajac.2015.66052.
References

[1]   http://www.rxlist.com/eliquis-drug.htm

[2]   Pujadas, M.L., Escolar, G., Arellano-Rodrigo, E. and Galan, A. (2013) Apixaban in the Prevention of Stroke and Systemic Embolism in Nonvalvular Atrial Fibrillation. Drugs Today (Barc), 49, 425-436.

[3]   Frost, C., Wang, J. and Nepal, S. (2013) Apixaban, an Oral, Direct Factor Xa Inhibitor: Single Dose Safety, Pharmacokinetics, Pharmacodynamics and Food Effect in Healthy Subjects. British Journal of Clinical Pharmacology, 75, 476-487.

[4]   Turpie, A.G. (2007) Oral, Direct Factor Xa Inhibitors in Development for the Prevention and Treatment of Thromboembolic Diseases. Arteriosclerosis Thrombosis and Vascular Biology, 27, 1238-1247.

[5]   Lassen, M.R., Davidson, B.L., Gallus, A., Pineo, G., Ansell, J. and Deitchman, D. (2007) The Efficacy and Safety of Apixaban, an Oral, Direct Factor Xa Inhibitor, as Thromboprophylaxis in Patients Following Total Knee Replacement. Journal of Thrombosis and Haemostasis, 5, 2368-2375.

[6]   Lassen, M.R., Gallus, A., Raskob, G.E., Pineo, G., Chen, D. and Ramirez, L.M. (2010) Apixaban versus Enoxaparin for Thromboprophylaxis after Hip Replacement. The New England Journal of Medicine, 363, 2487-2498.

[7]   Solanki, P.V., Uppelli, S.B., Dhokrat, P.V. and Mathad, V.T. (2015) Investigation on Polymorphs of Apixaban, an Anticoagulant Drug: Study of Their Transformations and Designing Efficient Process for Their Preparation. World Journal of Pharmaceutical Science, 3, 663-677.

[8]   Wren, S.A.C. and Tchelitcheff, P. (2006) Use of Ultra-Performance Liquid Chromatography in Pharmaceutical Development. Journal of Chromatography A, 1119, 140-146.

[9]   Snyder, L.R., Kirkland, J.J. and Dolan, J.W. (1997) Introduction to Modern Liquid Chromatography.

[10]   Cunliffe, J.M. and Maloney, T.D. (2007) Fused-Core Particle Technology as an Alternative to Sub-2-Micron Particles to Achieve High Separation Efficiency with Low Backpressure. Journal of Separation Science, 30, 3104-3109.

[11]   Samanidou, V.F. (2013) Core-Shell Particle Technology in Pharmaceutical Analysis. Pharmaceutica Analytica Acta, 4, e148.
http://dx.doi.org/10.4172/2153-2435.1000e148

[12]   http://www.phenomenex.com/Kinetex/CoreShellTechnology

[13]   Delavenne, X., Mismetti, P. and Basset, T. (2013) Rapid Determination of Apixaban Concentration in Human Plasma by Liquid Chromatography/Tandem Mass Spectrometry: Application to Pharmacokinetic Study. Journal of Pharmaceutical and Biomedical Analysis, 150, 78-79.

[14]   Pursley, J., Shen, J.X., Schuster, A., Dang, O.T., Lehman, J., Buonarati, M.H., Song, Y., Aubry, A.F. and Arnold, M.E. (2014) LC-MS/MS Determination of Apixaban (BMS-562247) and Its Major Metabolite in Human Plasma: An Application of Polarity Switching and Monolithic HPLC Column. Bioanalysis, 6, 2071-2082.

[15]   Gous, T., Couchman, L., Patel, J.P., Paradzai, C., Arya, R. and Flanagan, R.J. (2014) Measurement of the Direct Oral Anticoagulants Apixaban, Dabigatran, Edoxaban, and Rivaroxaban in Human Plasma Using Turbulent Flow Liquid Chromatography with High-Resolution Mass Spectrometry. Therapeutic Drug Monitoring, 36, 597-605.

[16]   Prabhune, S.S., Jaguste, R.S., Kondalkar, P.L. and Pradhan, N.S. (2014) Stability-Indicating High Performance Liquid Chromatographic Determination of Apixaban in the Presence of Degradation Products. Scientia Pharmaceutica, 82, 777-785.

[17]   The United States Pharmacopeia (2015) Validation of Compendial Methods. USP38 NF-33, Chapter 1225.

[18]   ICH Q2 (R1) (2005) Validation of Analytical Procedure: Text and Methodology. International Conference on Harmonization (ICH).

[19]   Ruan, J., Tattersall, P., Lozano, R. and Shah, P. (2006) The Role of Forced Degradation Studies in Stability Indicating HPLC Method Development. American Pharmaceutical Review, 9, 46-53.

[20]   Bakshi, M. and Singh, S. (2002) Development of Validated Stability-Indicating Assay Methods-Critical Review. Journal of Pharmaceutical and Biomedical Analysis, 28, 1011-1040.

[21]   Carstensen, J.T. and Rhodes, C.T. (2000) Drug Stability Principles and Practices. 3rd Edition, Marcel Dekker, New York.

 
 
Top