Parallel reaction monitoring MS

Parallel reaction monitoring mass spectrometry. Metabolic features having correlative trends to heat stress were first identified from the full-scan MS runs. These metabolic features were then subjected to parallel reaction monitoring (PRM) to generate high quality MS² spectra. We also collected the high quality MS² spectra from synthetic standards in order to confirm the metabolic features identified from the coral samples. For each dipeptide verified in the manuscript, multiple isomer matches existed based on the exact mass and key feature aspects as determined by the MS² spectra (Table below). The specific instrument settings of each PRM method event are include in the table below.

The MS² spectra generation was performed on a Thermo Q Exactive PLUS with a HESI source which was set to a spray voltage of -2.7kV under negative mode and 3.5kV under positive mode. The sheath, auxiliary, and sweep gas flow rates of 40, 10, and 2 (arbitrary unit) respectively. The capillary temperature was set to 300°C and aux gas heater was 360°C. The S-lens RF level was 45. The AGC target was set to 2e5 and the maximum IT was 100ms. The resolution was set to 70,000. The isolation window was set to 2.0 m/z. Collision energy levels were set to 30, 55 and 80 NCE.

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