Force degradation behavior of glucocorticoid deflazacort by UPLC: isolation, identification and characterization of degradant by FTIR, NMR and mass analysis.

Kesharwani Prashant,Trivedi Piyush,Tekade Muktika,Sharma Lata,K. Rakesh,Deshmukh Rajesh

doi:10.7555/jbr.30.20150074

Abstract

In this investigation, sensitive and reproducible methods are described for quantitative determination of deflazacort in the presence of its degradation product. The method was based on high performance liquid chromatography of the drug from its degradation product on reverse phase using Acquity UPLC BEH C18 columns (1.7 µm, 2.1 mm × 150 mm) using acetonitrile and water (40:60 V/V) at a flow rate of 0.2 mL/minute in UPLC. UV detection was performed at 240.1 nm. Deflazacort was subjected to oxidative, acid, base, hydrolytic, thermal and photolytic degradation. The drug was found to be stable in water and thermal stress, as well as under neutral stress conditions. However, forced-degradation study performed on deflazacort showed that the drug degraded under alkaline, acid and photolytic stress. The degradation products were well resolved from the main peak, which proved the stability-indicating power of the method. The developed method was validated as per ICH guidelines with respect to accuracy, linearity, limit of detection, limit of quantification, accuracy, precision and robustness, selectivity and specificity. Apart from the aforementioned, the results of the present study also emphasize the importance of isolation characterization and identification of degradant. Hence, an attempt was made to identify the degradants in deflazacort. One of the degradation products of deflazacort was isolated and identified by the FTIR, NMR and LC-MS study.

Highlights

Deflazacort is an oxazoline prednisolone derivative with the chemical structure of (11β,16β)-21-(acetyloxy)-11-hydroxy-2′-methyl-5′H-pregna-1,4-dieno [17,16-d] oxazole-3, 20-dione[1]
We have successfully developed a gradient UPLC method for the determination of deflazacort in presence of its degradants
The chromatographic separation method was optimized in Acquity UPLC BEH C18 column (1.7 μm, 2.1 mm × 150 mm), wherein, different mobile phases were investigated serially so as to find the best separation of pure deflazacort from its degradant products and obtain a clean chromatogram

Summary

Introduction

Deflazacort is an oxazoline prednisolone derivative with the chemical structure of (11β,16β)-21-(acetyloxy)-11-hydroxy-2′-methyl-5′H-pregna-1,4-dieno [17,16-d] oxazole-3, 20-dione[1]. It is a glucocorticoid with potent anti-inflammatory and immunosuppressant activity[2,3,4]. Administered deflazacort is well absorbed and immediately converted by plasma esterases to the pharmacologically active metabolite (D 21-OH), which achieves peak plasma concentrations in 1.5 to 2 hours[5,6]. Metabolism of D 21-OH is extensive and only 18% of urinary excretion represents D 21-OH[7]. The metabolite of D 21-OH, deflazacort 6-beta-OH represents one-third of urinary elimination[3,6]

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Conclusion