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Separation and determination of closely related lantibiotics by micellar electrokinetic chromatography
Authors:Bob Stanley  Kevin A. Mehr  Trent Kellock  Jonathan D. Van Hamme  Kingsley K. Donkor
Affiliation:1. Department of Chemistry, Thompson Rivers University, Kamloops, BC., Canada. Fax: +1‐250‐8285450;2. Department of Biological Sciences, Thompson Rivers University, Kamloops, BC., Canada
Abstract:A sensitive micellar electrokinetic chromatography (MEKC) method was developed for the separation and determination of four closely related lantibiotics: gallidermin, cinnamycin, duramycin and nisin. Factors affecting the separation of the lantibiotics such as pH, phosphate buffer concentration, SDS concentration and wavelength for UV detection were investigated. By optimizing these experimental conditions, successful separation was achieved between class 1A lantibiotics (nisin and gallidermin) and class 1B lantibiotics (duramycin and cinnamycin). The four lantibiotics were separated within 12 min in 50 mM phosphate buffer at pH 3.95 ± 0.1 containing 80 mM SDS with UV detection of 214 nm. The LOD (S/N = 3) were 61 ng/mL for gallidermin, 57 ng/mL for cinnamycin, 55 ng/mL for duramycin and 58 ng/mL for nisin. The method was successfully applied to real samples such as fermentation broth, bovine colostrum and predrop beer. This method yielded satisfactory results, with quantitative recoveries of spiked lantibiotics in the three samples ranging from 86.1 to 99.6%.
Keywords:Capillary electrophoresis  Cinnamycin  Duramycin  Gallidermin  MEKC  Nisin
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