High performance liquid chromatography (HPLC) method coupled with resonance Rayleigh scattering detection for the determination of isepamicin

Abstract

A simple and novel reversed-phase high performance liquid chromatography (HPLC) method coupled with resonance Rayleigh scattering (RRS) was developed to analyze isepamicin (ISP) in rat plasma. The chromatographic separation was achieved at 30 °C on a reverse phase column of Synergi Hydro-RP (150 mm × 4.6 mm; 4 μm), with (15 : 85, v/v) methanol–water (containing 0.15% TFA (v/v)) as the mobile phase. The flow rate was 0.4 mL min⁻¹. The RRS signal was detected at λ_ex = λ_em = 372 nm. Netilmicin (NTL) was used as the internal standard. The new analytical technique was validated for the intra- and inter-day variation, the linear range of the standard curve of ISP and the limit of detection (S/N = 3). The results indicated that HPLC coupled with RRS was a reliable and valuable technique for quantitative analysis of ISP in rat plasma. The method shows a better linear relationship and lower analysis time.