title: HPLC-ELSD determination of kanamycin B in the presence of kanamycin A in fermentation broth.
abstract: A novel method for the direct determination of kanamycin B in the presence of kanamycin A in fermentation broth using high performance liquid chromatography with evaporative light scattering detector (HPLC-ELSD) was developed. An Agilent Technologies C18 column was utilized, evaporation temperature of 40°C and nitrogen pressure of 3.5 bar, the optimized mobile phase was water-acetonitrile (65:35, v/v), containing 11.6 mm heptafluorobutyric acid (isocratic elution with flow rate of 0.5 mL/min) with the gain 11. Kanamycin B was eluted at 5.6 min with an asymmetry factor of 1.827. The method showed good linearity over the concentration range of 0.05 to 0.80 mg/mL for the kanamycin B (r(2) = 0.9987). The intra-day and inter-day coefficients of variation obtained from kanamycin B were less than 4.3%. Mean recovery of kanamycin B from spiked fermentation broth was 95%. The developed method was applied to the determination of kanamycin B without any interference from other constituents in the fermentation broth. This method offers simple, rapid and quantitative detection of kanamycin B.
authors.affiliation: State Key Laboratory of Chemical Resource Engineering, Beijing University of Chemical Technology, Beijing, 100029, People's Republic of China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hui-Min
authors.initials: HM
authors.lastname: He
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
authors.forename: Feng-Jiao
authors.initials: FJ
authors.lastname: Liu
authors.forename: Chao
authors.initials: C
authors.lastname: Li
authors.forename: Bing-Wu
authors.initials: BW
authors.lastname: Wang
authors.forename: Ren-Zhong
authors.initials: RZ
authors.lastname: Qiao
mesh_terms: D002138:Calibration; D005591:Chemical Fractionation / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D003470:Culture Media / Q000032:analysis / Q000737:chemistry; D004355:Drug Stability; D005285:Fermentation; D007612:Kanamycin / Q000031:analogs & derivatives* / Q000032:analysis / Q000302:isolation & purification; D008027:Light; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D013696:Temperature
chemical_list: D003470:Culture Media; C001495:bekanamycin; D007612:Kanamycin
keywords: HPLC-ELSD; fermentation broth; kanamycin; sample preparation