Figure 4.
Performance of high-pH peptide fractionation: (A) mass chromatograms of ENL T43 peptide recorded for 3 consecutive fractions during a 10-step 2D fractionation experiment: fraction 4/10 corresponds to a step elution from 14.0 to 15.4% ACN; fraction 5/10 was eluted from 15.4 to 16.7% ACN; fraction 6/10 (16.7 to 18.6% ACN). (B) mass chromatograms of ADH T26 peptide (EALDFFAR, [MH]1+ = 968.48) recorded for 3 consecutive fractions during a 10-step 2D fractionation experiment: fraction 5/10 corresponds to a step elution from 15.4 to 16.7% ACN; fraction 6/10 was eluted from 16.7 to 18.6% ACN; fraction 7/10 (18.6 to 20.4% ACN). The data was acquired in continuum mode and all mass chromatograms used an extraction window of 0.1 Da around the corresponding monoisotopic peaks. All separations were performed using a 30 min gradient (7–35% ACN, 0.1% FA). The amount loaded on-column was 20 fmoles ENL and 2 picomoles ADH digest.