Micromass quattro micro tandem mass spectrometer

The quantitative determination of tested compounds in cell lysates and culture media was performed on a Waters MICROMASS QUATTRO microTM tandem mass spectrometer combined with Acquity UPLC system. Acquity UPLC BEH C8 (2.1 × 50 mm × 1.7 μm) column was used at a flow rate of 0.25 mL/min. The column oven was set at 30 °C, and the sample injection volume was 5 μL. The mobile phase consisted of a combination of A (0.1% formic acid in water) and B (acetonitrile). The gradient elution program from 5% B to 98% B in 5 min was applied for analysis. Tandem mass spectrometer in positive electrospray mode was used for quantification. The TargetLynx software was applied to process LC/MS/MS data. Samples for LC/MS/MS analysis were prepared as follows: 200 μL of acetonitrile/methanol mixture (3:1 v/v) was add to 200 μL of each cell lysate and culture media sample. Then, the samples were centrifuged (10,000 rpm, 10 min), and the supernatants (300 μL) were removed from each sample and placed in vials for LC/MS/MS analysis. Calibration curves for all analytes were produced by analysis of standard solutions over a concentration range of 0.2 to 20 μM.

The quantitative determination of tested compounds in cell lysates and culture media was performed on a Waters MICROMASS QUATTRO microTM tandem mass spectrometer combined with Acquity UPLC system as described before [22 (link)].