COMPARATIVE STUDY OF FORCED DEGRADATION BEHAVIOR OF PRASUGREL BY UPLC AND HPLC AND THE DEVELOPMENT OF VALIDATED STABILITY INDICATING ASSAY METHOD

Abstract

A novel stability-indicating ultra-performance liquid chromatographic (UPLC) assay method was developed and validated for prasugrel and its degradation products. The UPLC separation was performed on Acquity® UPLC BEH C18 column (1.7 µm, 2.1 mm × 150 mm) using isocratic mode (acetonitrile:water, 80:20 v/v) at flow rate of 0.1 mL/min and the high performance liquid chromatography (HPLC) separation was achieved on Phenomenex® C8 column using isocratic mode (acetonitrile:10 mM ammonium acetate, 85:15 v/v) at flow rate of 0.9 mL/min. Prasugrel was found to degrade significantly in hydrolytic (acid, alkali, and neutral), and oxidative stress conditions and was stable in thermal and photolytic stress conditions. The RSD (%) values calculated for the AUC of UPLC and HPLC are 0.0039 and 0.0015, respectively. The UPLC and HPLC linearity of the proposed method were investigated in the range of 10–60 µg/mL. The r² value of UPLC and HPLC were found to be 0.9980 and 0.9983, respectively. Method detection limit (MDL) and method quantification limit (MQL) were found to be 0.20 µg/mL and 1.00 µg/mL for UPLC and 0.50 µg/mL and 1.80 µg/mL for HPLC, respectively. The RSD (%) values for intra-day and inter-day precision were <1.0%, confirming that the method is sufficiently precise. The validation studies were carried out fulfilling ICH requirements.

Keywords:

• HPLC
• prasugrel
• stability indicating assay
• stress testing
• UPLC
• validation

ACKNOWLEDGMENTS

The authors gratefully acknowledge the Vice Chancellor, RGPV, for providing the experimental facilities for this research work. The author Kapendra Sahu, wishes to thank AICTE, New Delhi, for postgraduate fellowships. One of the authors, C. Karthikeyan, wishes to thank CSIR, New Delhi, for providing a Senior Research Fellowship.

Notes

R_T = Retention time (minutes).

RR_T = Relative retention time.

SD = Standard deviation.

RSD = Relative standard deviation.

AUC = Area under curve.

SD = Standard deviation.

RSD = Relative standard deviation.

SD = Standard deviation.

RSD = Relative standard deviation.

UPLC = Ultra performance liquid chromatography.

HPLC = High performance liquid chromatography.

n = number of replicates.

R_T = Retention time (minutes).

RR_T = Relative retention time.

n = number of replicates.