Synapt g2 s quadrupole time of flight uplc q tof mass spectrometer

LC-MS analysis was performed using an ACQUITY ultra performance liquid chromatography (Waters Corp, Milford, MA) coupled with a Synapt G2-S quadrupole time-of-flight (UPLC-Q-TOF) mass spectrometer (Waters Corp, Wilmslow, UK). Samples were analysed on a BEH C₁₈ column (1.7 µm, 2.1 × 100 mm) which was maintained at 40 °C, whilst samples were maintained at 7 °C. Analytes were separated over a 15 min gradient using a flow rate of 0.5 mL/min. Mobile phase A was water and 0.1% formic acid, and B was methanol and 0.1% formic acid. The gradient consisted of initial conditions at 1% of B for 1 min, before increasing to 15% of B over 3 min, increasing to 50% of B over 3 min, and further increasing to 95% of B over 3 min and maintaining this for a further 1 min, before decreasing to 1% of B over 5 min, returning to initial conditions.
Column conditioning consisted of 8 repeat injections of the pooled QC. Samples were analysed as technical triplicates in a randomised order with the pooled QC injected every tenth injection throughout the analysis.
Cork plasma samples were analysed in December 2017 and Cork urine samples in May 2017. Auckland urine samples were analysed in July 2017. All samples were analysed on the same instrument, using a new BEH C₁₈ column for each experiment, following the protocols described above.

LC-MS analysis was performed using an ACQUITY ultra performance liquid chromatography (Waters Corp, Milford, MA) coupled with a Synapt G2-S quadrupole time-of-flight (UPLC-Q-TOF) mass spectrometer (Waters Corp, Wilmslow, UK). Samples were analysed on a BEH C₁₈ column (1.7 µm, 2.1 × 100 mm) which was maintained at 65 °C, whilst samples were maintained at 7 °C. Analytes were separated over a 23 min gradient using a flow rate of 0.4 mL/min. Mobile phase A was a mix of 10 mM of ammonium formate in acetonitrile:water (ACN:H₂O, 60:40 (v:v)), and B was a mix of 10 mM of ammonium formate in isopropanol:acetonitrile (IPA:ACN, 90:10 (v:v)). The gradient consisted of initial conditions at 30% of B, before increasing to 99% of B at 18 min and maintaining this for a further 2 min before decreasing to 30% of B over 2 min, returning to initial conditions. Column conditioning consisted of 8 repeat injections of the pooled QC. Samples were analysed as technical triplicates in a randomised order with the pooled QC injected every tenth injection throughout the analysis.