J. G. Phillips, Carole Simmonds
Jul 22, 1994
Citations
0
Influential Citations
15
Citations
Journal
Journal of Chromatography A
Abstract
An isocratic HPLC method was developed for the determination of spectinomycin dihydrochloride pentahydrate. The method is based on cation-exchange chromatography with pulsed-amperometric detection. The method was optimised for the separation of spectinomycin, actinamine and actinospectinoic acid on a Dionex IonPac CS3 column at ambient temperature with 150 mM sodium acetate (pH 6) mobile phase. Detector response with respect to sodium hydroxide concentration was investigated using cyclic voltammetry. A three-step pulse sequence with a sampling potential of + 0.05 V was used for detection. The k′ values for spectinomycin and related impurities were insensitive to column temperature changes in the range 21–50°C and show only minor variations to pH changes in the range 5.0–6.0. The method is linear and unbiased for the determination of spectinomycin over the concentration range 25–200 μg/ml for 50-μl injections. Two-day precision data for the method show within-sample R.S.D.s in the range 0.3–1.0% and between-day R.S.D.s in the range 0.3–0.9%. The limits of detection and quantitation determined for spectinomycin are 0.02 μg and 0.06 μg on-column, respectively. The method is suitable for the analysis of drug-substance.