| MS Description
|
Extraction of Metabolites:
The freeze-dri … Extraction of Metabolites:
The freeze-dried samples were extracted with 50 uL of 80% MeOH containing 2.5 uM lidocaine per mg dry weight using a mixer mill (MM300, Retsch) with zirconia beads for 10 min at 20 Hz and 4C. After centrifugation for 10 min, the supernatant was filtered using and HLB uElution plate (Waters).
LC-FTICR-MS Analysis:
A liquid sample (1 uL) was analyzed using LC-FTICR-MS (LC, Agilent 1200 series; MS, Bruker Daltonics solarix 7.0 T). Analytical conditions were as follows. LC: column, Xselect CSH C18 (3.5 um, 2.1 mm x 150 mm, Waters); solvent system, solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid); gradient program, 99.5% A/0.5% B at 0 min, 0.5% A/99.5% B at 30.0 min, 0.5% A/99.5% B at 45.min, 99.5% A/0.5% B at 45.1 min, and 99.5% A/0.5% B at 60.0 min; flow rate, 0.3 mL/min; column temperature, 35 C; wavelength, 200-600 nm; MS detection (resolution 260000 at m/z 400); polarity, positive; calibrant, low concentration TuningMix (Agilent); lock mass target, diethylhexylphthalate (C24H38O4, [M+H]+ m/z 391.28429); MS/MS detection (resolution 35000 at m/z 400). S detection (resolution 35000 at m/z 400).
|
