Journal Article

Development and Validation of a Stability-Indicating HPTLC-Based Assay for the Quantification of Nitrofurazone Ointment

Sikdar, K. M. Yasif Kayes, Lim, Lee Yong, Locher, Cornelia, Pacecca, Kate, Samie, Sarah, Islam, Md Khairul, Sostaric, Tomislav, Andrews, Hayley, Petker, Aliyah

2025

Overview

This paper reports on a validated, stability-indicating high-performance thin-layer chromatography (HPTLC)-based assay for the quantification of nitrofurazone in an ointment formulation. The simple and rapid HPTLC analysis was performed on silica gel 60 F254 HPTLC plates using toluene–acetonitrile–ethyl acetate–glacial acetic acid (6:2:2:0.1, v/v) as the mobile phase and chloroform–acetone (9:1, v/v) as the solvent. The method was validated in accordance with the guidelines set by both the International Council for Harmonisation (ICH) and the United States Food and Drug Administration (FDA). Nitrofurazone appeared as a sharp band with a RF value of 0.18. The method showed excellent linear regression between the concentration ranges of 30–180 ng/band (R = 99.99%). The limit of detection was found to be 10.39 ng/band, and the limit of quantification was 31.49 ng/band. The forced degradation of nitrofurazone via photolysis, oxidation, acid and alkaline hydrolyses confirmed the assay’s suitability for stability studies involving nitrofurazone. Therefore, the method is considered suitable for the routine quality control of nitrofurazone ointment.

• Publisher: MDPI AG,MDPI
• Subject: Acetic acid / Acetone / Acetonitrile / Analysis / Chromatography / Chromatography, High Pressure Liquid - methods / Chromatography, Thin Layer - methods / Drug Stability / Gram-positive bacteria / HPTLC / Limit of Detection / nitrofurazone / Nitrofurazone - analysis / Nitrofurazone - chemistry / ointment / Ointments - analysis / Ointments - chemistry / Organic compounds / Pharmaceuticals / Quality control / Quantitative analysis / Reproducibility of Results / Solvents / stability-indicating assay / Toluene / validation