title: Separation analysis of macrolide antibiotics with good performance on a positively charged C18HCE column.
abstract: The separation of basic macrolide antibiotics suffers from peak tailing and poor efficiency on traditional silica-based reversed-phase liquid chromatography columns. In this work, a C18HCE column with positively charged surface was applied to the separation of macrolides. Compared with an Acquity BEH C18 column, the C18HCE column exhibited superior performance in the aspect of peak shape and separation efficiency. The screening of mobile phase additives including formic acid, acetic acid and ammonium formate indicated that formic acid was preferable for providing symmetrical peak shapes. Moreover, the influence of formic acid content was investigated. Analysis speed and mass spectrometry compatibility were also taken into account when optimizing the separation conditions for liquid chromatography coupled with tandem mass spectrometry. The developed method was successfully utilized for the determination of macrolide residues in a honey sample. Azithromycin was chosen as the internal standard for the quantitation of spiramycin and tilmicosin, while roxithromycin was used for erythromycin, tylosin, clarithromycin, josamycin and acetylisovaleryltylosin. Good correlation coefficients (r(2) > 0.9938) for all macrolides were obtained. The intra-day and inter-day recoveries were 73.7-134.7% and 80.7-119.7% with relative standard deviations of 2.5-8.0% and 3.9-16.1%, respectively. Outstanding sensitivity with limits of quantitation (S/N ≥ 10) of 0.02-1 μg/kg and limits of detection (S/N ≥ 3) of 0.01-0.5 μg/kg were achieved.
authors.affiliation: School of Pharmacy, East China University of Science and Technology, Shanghai, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wei
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Aijin
authors.initials: A
authors.lastname: Shen
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Jingyu
authors.initials: J
authors.lastname: Yan
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Gaowa
authors.initials: G
authors.lastname: Jin
authors.affiliation: School of Pharmacy, East China University of Science and Technology, Shanghai, China.
authors.forename: Bingcheng
authors.initials: B
authors.lastname: Yang
authors.affiliation: Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Zhimou
authors.initials: Z
authors.lastname: Guo
authors.affiliation: School of Pharmacy, East China University of Science and Technology, Shanghai, China.
authors.forename: Feifang
authors.initials: F
authors.lastname: Zhang
authors.affiliation: School of Pharmacy, East China University of Science and Technology, Shanghai, China.|Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
authors.forename: Xinmiao
authors.initials: X
authors.lastname: Liang
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid; D006722:Honey / Q000032:analysis*; D018942:Macrolides / Q000032:analysis* / Q000302:isolation & purification*; D008968:Molecular Conformation; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry
chemical_list: D000900:Anti-Bacterial Agents; D018942:Macrolides
keywords: Macrolides; Peak shape; Positively charged surface; Reversed-phase liquid chromatography