DEVELOPMENT AND VALIDATION OF STABILITY-INDICATING REVERSE-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF POTENTIAL DEGRADATION PRODUCTS OF REGADENOSON FROM ITS PARENTERAL DOSAGE FORM
Objective: The objective was to develop and validate the stability indicating reverse-phase high-performance liquid chromatography method for the quantification of potential degradation products of regadenoson (REGA) from its injectable dosage form.
Methods: YMC-PAK ODS AQ, 150 mm Ã— 4.6 mm, 3 Î¼m composed with hydrophobic high carbon loading and a relatively hydrophilic surface chemically bonded to porous silica particles column was used with the temperature maintained at 40Â°C. Mobile phase A composed of 0.1% triethylamine buffer having pH 4.5 while mobile phase B is 100 % acetonitrile was used for gradient elution with 1.5 ml/min as a flow rate. The wavelength used for quantification was 245 nm and 20 Î¼l as an injection volume. The suitability of the method has been checked and validated according to the International Council for Harmonization (ICH) guidelines for different parameters, namely, specificity, linearity, accuracy, precision, limit of quantification (LOQ), Limit of detection (LOQ), and robustness studies.
Results: The resolution between REGA and its two-degradation product is >8.0 for all pairs of components. The high correlation coefficient (r2>0.990) values are for drug and all potential degradation products from LOQ to 150% of specification limits for impurities calculated based on the maximum daily dose of REGA. LOQ for the drug as well as each degradation product is <0.02% w/w. The % relative standard deviation (RSD) for precision and intermediate precision is in the range of 0.17â€“0.89, and % RSD for precision at LOQ is 0.86â€“2.35. The % RSD for robustness study is maximum 2.59.
Conclusion: The developed method can quantify the specified and unknown degradation products from 0.1% in the injectable dosage form which indicates that method is sensitive. Method fulfills the ICH criteria for its different validation parameters and demonstrates that the developed analytical method is highly specific, precise, and robust and would have a great value when applied in quality control and stability studies for REGA injection.
2. Rajesh BY, Appala Raju VV, Raju NA. A validated RP-HPLC method for the determination of 2-chloroadenosine as process related impurity in regadenosen parenteral dosage form. J Chem Pharm Res 2017;9:55-61.
3. Jackson S, Anders NM, Mangraviti A, Wanjiku TM, Sankey EW, Liu A, et al. The effect of regadenoson-induced transient disruption of the blood-brain barrier on temozolomide delivery to normal rat brain. J Neurooncol 2016;126:433-9.
4. ICH Q3B (R2). Impurities in New Drug Products. Geneva: International Conference on Harmonization; 2006.
5. Indian Express: Dr Reddyâ€™s, Aurobindo Recall Drugs From US Market; 2017. Available from: http://www.newindianexpress.com/ business/2017/feb/15/dr-reddys-aurobindo-recall-drugs-from-us-market-1570872.html. [Last accessed on 2017 Feb 15].
6. Vadlamudi MK, Dhanaraj S, Yarkala SA, Jayapal JJ, Kommavarapu PK. Development of stability indicating RP-HPLC method for the determination of related substances in atorvastatin solid dosage form and bulk drugs. Int J Pharm Pharm Sci 2015;7:184-94.
7. Mali AD, More UB. Development and validation of RP-HPLC method for simultaneous estimation of impurities from olmesartan medoximil and hydrochlorothiazide tablet. Int J Pharm Pharm Sci 2016;8:45-8.
8. United States Pharmacopeial Convention. The United States Pharmacopeia, USP 34-NF29. Asian edition. Rockville, MD: United States Pharmacopeial Convention; 2011. p. 2025-8.
9. The Stationery Office Publications. British Pharmacopeia. Vol. 1. London, UK: The Stationery Office Publications; 2011. p. 265.
10. Council of Europe. European Pharmacopeia. 5thed. Strasbourg: Council of Europe; 2011. p.1091
11. (ICH) guidelines Q2 (R1). Validation of Analytical Procedures: Text and Methodology. Geneva, Switzerland: (ICH) guidelines Q2 (R1); 2005.
12. (ICH) guidelines Q2A (R2). Stability Testing of New Drug Substances and Product. Geneva, Switzerland: (ICH) guidelines Q2 (R1); 2003.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.