Separation and determination of closely related lantibiotics by micellar electrokinetic chromatography.
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%.
Stanley
Bob
B
Department of Chemistry, Thompson Rivers University, Kamloops, BC., Canada.
Mehr
Kevin A
KA
Kellock
Trent
T
Van Hamme
Jonathan D
JD
Donkor
Kingsley K
KK
D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification; D001430:Bacteriocins / Q000737:chemistry / Q000302:isolation & purification; D001515:Beer; D002417:Cattle; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000295:instrumentation / Q000379:methods; D003126:Colostrum / Q000737:chemistry; D008969:Molecular Sequence Data; D009561:Nisin / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D010456:Peptides, Cyclic / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D010455:Peptides; D010456:Peptides, Cyclic; C057656:gallidermin; C040522:duramycin; C000485:cinnamycin; D009561:Nisin