Development and Validation of a Stability Indicating Rp-HPLC Method for the Determination of Potential Degradation Products of Difluprednate in Ophthalmic Emulsion

Abstract

Objective: The objective of the current study was to develop and validate a simple, robust, precise and accurate RP-HPLC (reverse phase-high performance liquid chromatography) method for the quantitative determination of potential degradation products of Difluprednate (DIFL) in the ophthalmic emulsion.Methods: Chromatographic separation was achieved on the YMC pack ODS-AQ (150× 4.6) mm, 3μm column with a mobile phase containing a gradient mixture of mobile phase A (0.02M Ammonium formate buffer pH 4.5 adjusted with formic acid) and Acetonitrile as mobile phase B, at flow rate of 1.5 ml/min and with UV detection at 240 nm.Results: The peak retention time of DIFL was found at about 17.2 min, the RRT of degradation product-1 (DP-1), degradation product-2 (DP-2), and degradation product-3 (DP-3), were found to be about 0.49, 0.65 and 0.79 respectively (calculated with respect to Difluprednate). Stress testing was performed in accordance with an ICH (international council for harmonisation) guideline Q1A (R2) [1]. The method was validated as per ICH guideline Q2 (R1)[2]. The calibration curve was found to be linear in the concentration range of 0.1 to 0.75 µg/ml for Difluprednate, DP-1, DP-2 and DP-3. The LOD (Limit of detection) was found to be 0.1µg/ml and LOQ (Limit of quantification) of 0.15µg/ml for Difluprednate, DP-1, DP-2 and DP-3 respectively. The recovery from LOQ to 150% was within 90-110%. The forced degradation data confirms the stability indicating the nature of the method.Conclusion: A simple, robust, precise and accurate RP-HPLC method for the quantitative determination of potential degradation products of Difluprednate in the ophthalmic emulsion was developed and validated.

Hydroxyapatite and Silicon-Modified Hydroxyapatite as Drug Carriers for 4-Aminopyridine

“…Its mechanism of action lies in its potential to block the voltage-dependent potassium channels (Kv1), contributing to the improvement of the action potential propagation in demyelinated axons [28,29]. The extensive literature survey concerning the analytical methods that have been developed so far for the analysis of 4AP implies the use of HPLC methods [30][31][32][33][34], LC-MS techniques [35][36][37], as well as simple UV-spectroscopic determinations [38,39]. A reference method for the in vitro release kinetics of the active substance has been disclosed by the FDA, which uses the paddle method, at 50 rpm, in 900 mL phosphate buffer solution at pH = 6.8, with specific sampling time points of 0.5, 1, 2, 4, 6, 8, 10, and 12 h [40].…”

Quality by Design Approach to Stability-Indicating Reverse-Phase High-Performance Liquid Chromatography Method Development, Optimization, and Validation for the Estimation of Simeprevir in Bulk Drug

“…Hence, the quality by design (QbD) paradigm's principles have been utilized to develop a successful stability-indicating method. The International Conference on Harmonization (ICH) Q8 (R1) guideline defines QbD as "a systematic approach to development that begins with predefined objectives and emphasizes product and process understanding and process control, based on sound science and quality risk management" [2,12,13].…”