| MS Description
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Extraction of Metabolites<br />
The … Extraction of Metabolites<br />
The freeze-dried samples were extracted in a mixer mill (MM300, Retsch) with 50 μL of 80% MeOH per mg dry weight and zirconia beads. After 7 min of milling at 18 Hz and 4 °C, the extractions were centrifuged for 10 min and the supernatant was filtered through an HLB μElution plate (Waters).<br /><br />
LC–FTICR–MS Analysis<br />
Ultrahigh-resolution metabolome data were acquired by an FTICR–MS solariX 7.0 T (Bruker Daltonics) with the ESI source. LC–FTICR–MS analysis was performed as previously described. The FTICR–MS was controlled by the software ftmsControl 2.1.0 (Bruker Daltonics). For internal calibration, lidcaine (250 μM in MeOH; Tokyo Chemical Industry Co. Ltd., Tokyo, Japan) was added to both solvent A (water with 0.1% formic acid, Wako Pure Chemical Industries Ltd.) and solvent B (acetonitrile with 0.1% formic acid, Wako Pure Chemical Industries Ltd.). The column was changed to an Xselect CSH Phenyl-Hexyl (3.5 μm, 2.1 mm × 150 mm, Waters, Milford, MA, U.S.A.).<br />
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LC–FTICR–MS/MS Analysis<br />
The MS/MS boost analysis was carried out at a collision energy of 30 V as previously described. on energy of 30 V as previously described.
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