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Sampling methods for NMR‐based metabolomics of Staphylococcus aureus
Author(s) -
Wu XiaoHe,
Yu HaiLong,
Ba ZhaoYong,
Chen JingYu,
Sun HongGang,
Han BeiZhong
Publication year - 2010
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200900038
Subject(s) - reproducibility , metabolite , metabolomics , staphylococcus aureus , quenching (fluorescence) , chromatography , yield (engineering) , filtration (mathematics) , chemistry , biology , biochemistry , bacteria , materials science , mathematics , physics , statistics , quantum mechanics , metallurgy , fluorescence , genetics
To select an appropriate sampling method for comparison of metabolite profiles between planktonic and biofilm Staphylococcus aureus using NMR techniques, we evaluated three methods: quenching‐centrifugation (QC), filtration‐quenching (FQ) and filtration‐quenching‐lyophilization (FQL). We found differences in metabolite loss, yield, reproducibility and metabolite profile. QC caused severe metabolite leakage and possible decomposition of nucleotides. FQ achieved high yields and reproducibility, although it had the disadvantages of long filtration and rinse times before quenching. FQL resulted in a loss of a few metabolites and a lower yield due to lyophilization. Although the biomarkers discovered by each method were nearly the same and seemed insensitive to technical variances, we conclude that FQ is the most appropriate sampling method because of its high yield and reproducibility.