|SE57 > MS01|
|Title||Acquisition of ESI-MS and ESI-MS/MS data, UPLC-TQMS analysis|
|Instrument||Waters Xevo TQD, UPLC (Waters)|
|Description||<Acquisition of ESI-MS and ESI-MS/MS data>
All the solutions of authentic compounds (250 μ M) were transferred from 10 ml vials to 1.2 ml glass inserts in 96-well plates (Webseal system, GL Sciences, Tokyo, Japan) and diluted to 50 pmol μ l –1 with H 2 O using an ALHS (Microlab STARplus, Hamilton, Reno, NV, USA). The solutions (20 μ l aliquots containing 1 nmol of compound) were analyzed by the flow injection method using a CTC-PAL injection system (AMR, Tokyo, Japan) and a Waters GI Pump solvent system (Waters, Milford, MA, USA) consisting of: 0.05 ml min –1 flow using a micro splitter (GL Sciences), 80% acetonitrile (0.1% formic acid) and 20% water (0.1% formic acid), with a 2.5 min cycle in the isocratic mode. The ionized authentic compounds were detected by TQMS (TQD, Waters) according to the following conditions: capillary voltage +3.0 keV or –2.8 keV, CV 10–60 eV (six levels), source temperature 120°C, desolvation temperature 350°C, cone gas fl ow 50 l h –1 , desolvation gas fl ow 600 l h –1 and CE 10–60 eV (six levels). A total of 61,920 spectra of 860 compounds were obtained.The optimal MRM conditions, including positive/negative polarity (e.g. [M] + , [M + H] + , [M] – , [M – H] – ), m / z of precursor ion and product ion, and optimal CV and CE were determined automatically for 497 of these compounds using the software QuanOptimize (Waters).
The UPLC (Waters) conditions were manually optimized based on the separation patterns of 12 methionine-derived glucosinolates and were as follows: flow rate 0.24 ml min –1 ; solvents A, 0.1% formic acid in water and B, 0.1% formic acid in acetonitrile; gradient program of B (0 min, 0%; 0.25 min, 0%; 0.4 min, 9%; 0.8 min, 17%; 1.9 min, 100%; 2.1 min, 100%;
2.11 min, 0%); 3 min cycles with a temperature of 38°C.The TQMS detection conditions were the same as those for FIA-TQMS, except that the source temperature was 130°C.