Ultimate 3000rs with tsq quantis ms ms detector

This analysis was performed on a UHPLC-DAD-MS/MS apparatus (Ultimate 3000RS with TSQ Quantis MS/MS detector; Thermo Fischer Scientific, Waltham, MA, USA) using a Hypersil GOLD column (3.0 µm, 3.0 × 100 mm; Thermo Fischer Scientific, Waltham, MA, USA). A gradient consisting of solvent A (50 μM NaCl in H₂O) and solvent B (acetonitrile:H₂O, 30:70, v/v) was applied at a flow rate of 0.5 mL/min as follows: 0.14 min, 96% A and 4% B; 7.84 min, 14% A and 86% B; 8.96 min, 14% A and 86% B; 9.52 min, 5% A and 95% B; 13.16 min, 5% A and 95% B; 13.44 min, 96% A and 4% B; and 15.68 min, 96% A and 4% B. The column temperature was held at 25 °C, and the injection volume was 5 µL. The system was operated in the positive ion electrospray mode, with the electrospray interface H-ESI operating at a capillary voltage of 3.5 kV and a temperature of 350 °C. The signals were recorded at 227 nm by a DAD. dGSLs were quantified using an external calibration curve of pure desulfosinigrin (range from 13.56 to 542.50 µM). For each individual dGSL, response factors (RPFs) were taken into account in accordance with the literature as follows: RPF 1.0 for 1, 1.1 for 3 [45 (link)], 0.95 for 4, 0.29 for 5, and 0.25 for 6 [46 ]; arbitrary 1.0 for 2.

Analysis was performed on UHPLC-DAD-MS/MS (Ultimate 3000RS with TSQ Quantis MS/MS detector, Thermo Fischer Scientific, Waltham, MA, USA) using Hypersil GOLD column (3.0 µm, 3.0 × 100 mm, Thermo Fischer Scientific). A gradient consisting of solvent A (50 μM NaCl in H₂O) and solvent B (acetonitrile:H₂O 30:70 v/v) was applied at a flow rate of 0.5 mL/min as follows: 0.14 min 96% A and 4% B; 7.84 min 14% A and 86% B; 8.96 min 14% A and 86% B; 9.52 min 5% A and 95% B; 13.16 min 5% A and 95% B; 13.44 min 96% A and 4% B; 15.68 min 96% A and 4% B. The column temperature was held at 25 °C and the injection volume was 5 µL. The electrospray interface was H-ESI source operating with a capillary voltage of 3.5 kV at 350 °C. The system was operated in the positive ion electrospray mode.
The amount of GSLs was quantified using a calibration curve (y = 0.0206x + 0.2371, R² = 0.9992, LOD = 1.67 μM, LOQ = 5.03 μM) of pure desulfosinigrin solution (range from 13.63 to 545.00 µM) and RPFs for each individual desulfo-GSL [33 (link)]. RPF values for quantification of desulfo-GSLs were as follows: RPF 1.09 for 1 and 3, 1.07 for 2, 1.11 for 4, 1.04 for 5, 0.50 for 6, 0.95 for 9 and 13 [34 ]; RPF 0.55 for 12 [7 (link)]; arbitrary RPF 1.0 for 7, 8, 10 and 11.

Analysis was performed on UHPLC-MS/MS (Ultimate 3000RS with TSQ Quantis MS/MS detector, Thermo Fischer Scientific, USA) using Hypersil GOLD column (3.0 µm, 3.0 × 100 mm, Thermo Fischer Scientific, USA). A gradient consisting of solvent A (50 μM NaCl in H₂O) and solvent B (acetonitrile/H₂O 30:70 v/v) was applied at a flow rate of 0.5 mL/min as follows: 0.14 min 96% A and 4% B, 7.84 min 14% A and 86% B, 8.96 min 14% A and 86% B;,9.52 min 5% A and 95% B, 13.16 min 5% A and 95% B, 13.44 min 96% A and 4% B, and 15.68 min 96% A and 4% B. The column temperature was held at 40 °C and the injection volume was 5 µL. The electrospray interface was H-ESI source operating with a capillary voltage of 3.5 kV at 350 °C. The system was operated in the positive ion electrospray mode.