Disaccharide analysis was performed according to a previously reported method [37 (link)]. LC was performed on an Agilent 1200 LC system at 45 °C using an Agilent Poroshell 120 ECC18 (2.7 μm, 3.0 × 50 mm) column. Mobile phase A (MPA) was 50 mM ammonium acetate aqueous solution, and the mobile phase B (MPB) was methanol. The mobile phase passed through the column at a flow rate of 300 μL/min. The gradient used was the following: 0-10 min, 5-45% B; 10-10.2 min, 45-100% B; 10.2-14 min, 100% B; 14-22 min, 100-5% B. The injection volume used for all the samples was 5 μL.
A triple quadrupole mass spectrometry system equipped with an ESI source (Thermo Fisher Scientific, San Jose CA, USA) was used a detector. The online MS analysis was performed at the Multiple Reaction Monitoring (MRM) mode with the MS parameters: negative ionization mode with a spray voltage of 3000 V, a vaporizer temperature of 300 °C, and a capillary temperature of 270 °C. Data analysis was performed using Thermo Xcalibur™ software (Thermo Fisher Scientific, San Jose CA, USA). The disaccharides in different cell and cell-derived ECM samples were quantified by comparison of the sample peak area to that of an external standard.
A triple quadrupole mass spectrometry system equipped with an ESI source (Thermo Fisher Scientific, San Jose CA, USA) was used a detector. The online MS analysis was performed at the Multiple Reaction Monitoring (MRM) mode with the MS parameters: negative ionization mode with a spray voltage of 3000 V, a vaporizer temperature of 300 °C, and a capillary temperature of 270 °C. Data analysis was performed using Thermo Xcalibur™ software (Thermo Fisher Scientific, San Jose CA, USA). The disaccharides in different cell and cell-derived ECM samples were quantified by comparison of the sample peak area to that of an external standard.
