Sampling methods for NMR-based metabolomics of Staphylococcus aureus

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 diffe...

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Bibliographic Details
Published inBiotechnology journal Vol. 5; no. 1; p. 75
Main Authors Wu, Xiao-He, Yu, Hai-Long, Ba, Zhao-Yong, Chen, Jing-Yu, Sun, Hong-Gang, Han, Bei-Zhong
Format Journal Article
LanguageEnglish
Published Germany 01.01.2010
Subjects
Algorithms
Bacterial Proteins - metabolism
Data Interpretation, Statistical
Gene Expression Profiling - methods
Magnetic Resonance Spectroscopy - methods
Metabolome - physiology
Proteome - metabolism
Specimen Handling - methods
Staphylococcus aureus - metabolism
Ultrafiltration - methods
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Summary: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.
ISSN:1860-7314
DOI:10.1002/biot.200900038