Stability-indicating high performance thin layer chromatographic determination of sulfanilamide in human urine

Abstract

A simple, sensitive, selective, precise and stability-indicating high-performance thin-layer chromatographic (HPTLC) method was developed and applied to human urine for the densitometric determination of sulfanilamide. A mixture of chloroform-ethyl acetate-xylene (2.5: 4.0: 1.0, v/v/v) was used as a mobile phase. The system was found to give compact spots for sulfanilamide (retardation factor, R f = 0.21±0.02). The linear regression analysis data for the calibration plots showed good linear relationship with r 2 = 0.9970 ± 0.0003 and r 2 = 0.9947 ± 0.020 within the concentration range of 50–250 ng per spot and 100–1000 ng per spot with respect to peak area, respectively. The limit of detection (LOD) and quantification (LOQ) were 8 and 25 ng per spot, respectively. Sulfanilamide was subjected to acid and alkali hydrolysis, oxidation, dry heat and wet heat treatment. According to the International Conference on Harmonization (ICH) guidelines the method was validated for precision, recovery and robustness. The ultraviolet (UV) spectra of the degradation products which had different spectra from sulfanilamide were also recorded.