DEVELOPMENT AND VALIDATION OF STABILITY INDICATING RP-HPLC METHOD FOR DETERMINATION OF Î²-ACETYLDIGOXIN
Objective: The objective of the present study was to develop and validate a novel stability indicating reverse phase-high performance liquid chromatography (RP-HPLC) method for determination of Î²-acetyldigoxin, an active pharmaceutical ingredient (API).
Methods: The chromatographic separation was carried out on Agilent Technologies 1200 series HPLC system equipped with photo diode array detector and C-18 (4.6x250 mm, 5 Âµ) column. The mobile phase consisted of water: acetonitrile (65:35 v/v), delivered at a flow rate of 1.5 ml/min and eluents were monitored at 225 nm.
Results: The retention time of Î²-acetyldigoxin was 9.2 min. The method was found to be linear (R2= 0.9995) in the range of 31.25-500 Âµg/ml. The accuracy studies showed the mean percent recovery of 101.02%. LOD and LOQ were observed to be 0.289 Âµg/ml and 0.965 Âµg/ml, respectively. The method was found to be robust and system suitability testing was also performed. Forced degradation analysis was carried out under acidic, alkaline, oxidative and photolytic stress conditions. Significant degradation was observed under tested conditions, except for oxidative condition. The method was able to separate all the degradation products within runtime of 20 min and was able to determine Î²-acetyldigoxin unequivocally in presence of degradation products.
Conclusion: The novel, economic, rapid and simple method for analysis of Î²-acetyldigoxin is reported. The developed method is suitable for routine quality control and its determination as API, and in pharmaceutical formulations and stability study samples.
2. Julie HW. Digoxin toxicity: a review. US Pharm 2006;2:28-36.
3. Estevao LF, Fabricio PM. Digoxinâ€™s roles in heart failure patients: an overview. Insuficiencia Cardiaca 2010;5:1022-32.
4. Cheng JWM, Rybak I. Use of digoxin for heart failure and atrial fibrillation in elderly patients. Am J Geriatr Pharmacother 2010;5:419-27.
5. Flasch H, Schumpelic V, Koch G. Fate of orally administered beta-acetyldigoxin in man. Arzneimittel Forschung 1976; 27:656-9.
6. Pauli-Magnus C, MÃ¼rdter T, Godel A, Mettang T, Eichelbaum M, Klotz U, et al. P-glycoprotein-mediated transport of digitoxin, Î±-methyldigoxin and Î²-acetyldigoxin. Naunyn-Schmiedeberg's Arch Pharmacol 2001;363:337-43.
7. Flasch H. Bioavailability of Î²-acetyldigoxin und Digoxin. Klin Wochenschr 1975;53:873-7.
8. Gruppillo P, Padovan GC, Tomaini MD, Masoni C. Therapeutic comparison between digoxin, beta-methyl-digoxin and beta-acetyl-digoxin. La Clinica Terapeutica 1981;99:281.
9. Marcus FI. Pharmacokinetic interactions between digoxin and other drugs. J Am Coll Cardiol 1985;5:82-90.
10. Degner FL, Heinzel G, Narjes H, Turck D. The effect of meloxicam on the pharmacokinetics of betaâ€acetylâ€digoxin. Br J Clin Pharmacol 1995;40:486-8.
11. World Health Organization. IARC monographs on the evaluation of carcinogenic risks to humans. WHO 2016;108:381-419.
12. Thies PW. The modification of natural substances in the modern drug synthesis. In: New natural products and plant drugs with pharmacological, biological or therapeutical activity. Springer Berlin Heidelberg; 1977. p. 266-83.
13. Bakshi M, Singh S. Development of validated stability-indicating assay methods-critical review. J Pharm Biomed Anal 2002;28:1011-40.
14. Rizwan SH, Sastry G. Development and validation of stability indicating RP-HPLC method for the simultaneous estimation of aliskiren hemifumarate and valsartan in bulk and pharmaceutical dosage form. Asian J Pharm Clin Res 2015;8:223-7.
15. Plum J, Daldrup T. Detection of digoxin, digitoxin, their cardioactive metabolites and derivatives by high-performance liquid chromatography and high-performance liquid chromatography-radioimmunoassay. J Chromatogr B 1986;377:221-31.
16. Desta B, Kwong E, McErlane KM. Separation of digoxin, digitoxin and their potential metabolites, impurities or degradation products by high-performance liquid chromatography. J Chromatogr A 1982;240:137-43.
17. Kazakevich YV, LoBrutto R. editors. HPLC for Pharmaceutical Scientists. Wiley Publishers; 2007. p. 788.
18. International Conference on Harmonization (ICH), Q2(R1): Validation of analytical procedures: text and methodology; 2005.
19. Chapter G. Validation of compendial methods, United States Pharmacopeia. The United States Pharmacopoeia Convention. Rockville, MD; 2003.
20. FDA, Guidance R. Validation of chromatographic methods. Center for Drug Evaluation and Research (CDER), Food and Drug Administration; 1994. p. 2.
21. Jain HK, Deore DD. Bioanalytical method development and validation for estimation of clopidogrel bisulfate in human plasma by RP-HPLC. Int J Appl Pharm 2016;8:18-21.
22. Ahuja S, Dong M. editors. Handbook of Pharmaceutical Analysis by HPLC. 1st ed. USA: Elsevier-Academic Press; 2005.
23. FDA, ORA Validation and Verification Guidance for Human Drug Analytical Methods. Food and Drug Administration; 2003. p. 1.
24. FDA, Department of Health and Human Services, 510 (k) Summary Elecsys Progesterone III Calset, Food and Drug Administration; 2015. p. 45.