Characterization of the Oxidative Degradation Product of Darunavir by LC-MS/MS.

Karthik Yamjala,Nivedeetha Halekote Shivaraju ,Jeevitha Atukuri ,Krishnaveni Nagappan ,Meyyanathan Subramania Nainar

doi:10.3797/scipharm.1505-10

Abstract

A rapid, selective, and reliable LC-MSn method has been developed and validated for the isolation and structural characterization of the degradation product of darunavir (DRV). DRV, an HIV-1 protease inhibitor, was subjected to intrinsic oxidative stress conditions using 30% hydrogen peroxide and the degradation profile was studied. The oxidative degradation of DRV resulted in one degradation product. The unknown degradation product was separated on a Hibar Purospher C18 (250 mm × 4.6 mm; 5 µm) column by using 0.01 M ammonium formate (pH 3.0) and acetonitrile as mobile phase in the ratio of 50:50, v/v. The eluents were monitored at 263 nm using a UV detector. The isolated degradation product was characterized by UPLC-Q-TOF and its fragmentation pathway was proposed. The proposed structure of the degradation product was confirmed by HRMS analysis. The developed stability-indicating LC method was validated with respect to accuracy, precision, specificity/selectivity, and linearity. No prior reports were found in the literature about the oxidative degradation behavior of DRV.

Highlights

The term “quality of pharmaceuticals” from the previous focus on purity is being changed to have a greater emphasis on impurities and degradation products
Ammonium formate buffer (0.01 M, pH 3.0 adjusted with formic acid) /acetonitrile (50:50, v/v) in an isocratic mode and the Hibar Purospher C18 column (250 × 4.6 mm id, 5 μm) with stationary phase was used for the successful separation of DRV from its degradation product
The flow rate was 1.0 mL/min and the detection was carried out at 263 nm with 15 min of run time. These optimized conditions were used for the separation of DRV and its Characterization of the Oxidative Degradation Product of Darunavir by LC-MS/MS

Summary

Introduction

The term “quality of pharmaceuticals” from the previous focus on purity is being changed to have a greater emphasis on impurities and degradation products. K. Yamjala et al.: pharmaceuticals is always a balance between risks and benefits, but the same is not true for impurities in pharmaceuticals; impurities carry only risks [1]. Stress testing has long been recognized as an important part in the drug development process. The focus on these degradation products and impurities is basically due to various health and safety considerations. A subset of these impurities may cause potential genotoxicity, which poses an additional safety concern leading to carcinogenicity and genetic mutations

Objectives

Methods

Results