UHP-HILIC-MS analysis was performed using an
1290 Infinity UHPLC system (Agilent Technologies, Waldbronn, BW, Germany) consisting of a binary pump, an autosampler and a temperature controlled column department at 65°C with a Waters (Milford, MA, USA) acquity UPLC BEH amide 1.7 µM 3.0x100 mm HILIC column coupled to a
Q-TOF II mass spectrometer with an electrospray ionization source and liquid chromatography sprayer from Bruker Daltonics (Bremen, HB, Germany), operated in positive mode. The compounds were eluted isocratically with 40% water containing 300 µM formic acid and 550 µM NH 4 OH (pH 9.2) and 60% acetonitrile (% v/v) at a flow rate of 0.6 mL/min. The injection volume was 20 µL. MS settings were optimized for MNA signal-to-noise ratio. The optimal settings were a capillary voltage of 1500 V, a nebulizer pressure of 4.1 bar, a drying gas flow rate of 9.8 L/min, a drying temperature of 220 °C, a scan range of m/z 90-400 and a spectra sample rate of 3 spectra/s.
van Haren M.J., Sastre Toraño J., Sartini D., Emanuelli M., Parsons R.B, & Martin N.I. (2016). A Rapid and Efficient Assay for the Characterization of Substrates and Inhibitors of Nicotinamide N-Methyltransferase. Biochemistry, 55(37).