6495 triple quad ls ms

Glycolate was measured on a triple quadrupole mass spectrometer (Agilent Technologies 6495 Triple Quad LS–MS) equipped with UHPLC (Agilent Technologies 1290 Infinity II) using a 150 × 2.1 mm C18 column (Kinetex 1.7 µm EVO C18 100 Å) at 25 °C. The injection volume was 1 µl. The diluted samples (1:10 in water) as well as the external standard curve were diluted 1:2 with 10 µM ¹³C-labeled glycolate as the internal standard. The flow was set to 0.100 ml min⁻¹, and the separation was performed using dH₂O with 0.1% formic acid (buffer A) and methanol with 0.1% formic acid (buffer B). The gradient for LC–MS analysis and the parameters for the MRMs are displayed in Supplementary Tables 6 and 7, respectively. Data analysis was done using Agilent MassHunter Workstation Software.

Malate was measured on a triple-quadrupole mass spectrometer (Agilent Technologies 6495 Triple Quad LS–MS) equipped with UHPLC (Agilent Technologies 1290 Infinity II) using a 150 × 2.1 mm C18 column (Kinetex 1.7 µm EVO C18 100 Å) at 25 °C. The injection volume was 5 µl. The diluted samples (1:10 in water) as well as the external standard curve were diluted 1:2 with 10 µM ¹³C-labeled malate as the internal standard. The flow was set to 0.150 ml min⁻¹, and the separation was performed using dH₂O with 0.1% formic acid (buffer A) and methanol with 0.1% formic acid (buffer B). The gradient for LC–MS analysis and the parameters for the MRMs are displayed in Supplementary Tables 8 and 9, respectively. Data analysis was done using Agilent MassHunter Workstation Software.

All CoA esters were measured on a triple quadrupole mass spectrometer (Agilent Technologies 6495 Triple Quad LS–MS) equipped with UHPLC (Agilent Technologies 1290 Infinity II) using a 50 × 2.1 mm C18 column (Kinetex 1.7 µm EVO C18 100 Å) at 25 °C. The injection volume was 2 µl of the diluted samples (1:10 in water). The flow was set to 0.250 ml min^-1, and the separation was performed using 50 mM ammonium formate pH 8.1 (buffer A) and acetonitrile (buffer B). We quantified the CoA esters using external standard curves prepared in the 1:10 diluted (water) sample matrix. The gradient for LC–MS analysis and the parameters for the multiple reaction monitoring (MRMs) are displayed in Supplementary Tables 4 and 5, respectively. Data analysis was done using Agilent MassHunter Workstation Software.