title: Validation of a HILIC Method for the Analysis of Ergothioneine in Fermentation Broth.
abstract: A hydrophilic interaction liquid chromatography method has been established for the quantification of ergothioneine (EGT) in fermentation broth. Chromatographic separation was conducted on a Venusil hydrophilic interaction liquid chromatography (HILIC) column (250 × 4.6 mm, 5 µm) at an elution rate of 1.0 mL/min with an isocratic mobile phase consisting of acetonitrile/20 mmol/L ammonium acetate solution (85 : 15, v/v) adjusted to pH 6.0 with acetic acid. Analytes were detected at 254 nm using a UV-VIS detector. The injection volume was 10 µL, and the column temperature was 40°C. The limits of detection and limits of quantification were 63 and 21 µg/L, respectively. Excellent linearity [correlation coefficient (R(2)) = 0.9999] was achieved for EGT quantification in the range of 5-400 mg/L. The relative standard deviations of repeatability, intermediate precision and stability were 1.47, 1.03 and 1.66%, respectively, and EGT recoveries were within 99.2-100.8%. The chromatographic peak corresponding to EGT in the HILIC spectrum was confirmed using ESI-MS. In general, the method developed here is simple, reliable, accurate, and stable and may be useful for routine analyses in EGT biosynthesis research.
authors.affiliation: Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, PR China.
authors.forename: Qi
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, PR China College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300222, PR China.
authors.forename: Weiya
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Bonna-Agela Technologies Inc, Tianjin 300457, PR China.
authors.forename: Hongyu
authors.initials: H
authors.lastname: Wang
authors.affiliation: Bonna-Agela Technologies Inc, Tianjin 300457, PR China.
authors.forename: Yunhua
authors.initials: Y
authors.lastname: Li
authors.affiliation: Bonna-Agela Technologies Inc, Tianjin 300457, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.affiliation: Bonna-Agela Technologies Inc, Tianjin 300457, PR China.
authors.forename: Qunjie
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, PR China.
authors.forename: Dongze
authors.initials: D
authors.lastname: Liu
authors.affiliation: College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300222, PR China.
authors.forename: Ning
authors.initials: N
authors.lastname: Chen
authors.affiliation: Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Key Laboratory of Systems Microbial Biotechnology, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, PR China jiang_wx@tib.cas.cn.
authors.forename: Wenxia
authors.initials: W
authors.lastname: Jiang
mesh_terms: D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media / Q000737:chemistry*; D004880:Ergothioneine / Q000032:analysis*; D005285:Fermentation; D057230:Limit of Detection; D015203:Reproducibility of Results
chemical_list: D003470:Culture Media; D004880:Ergothioneine