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The column was eluted for 2 min from 0% B to 2% B (linear), from 3 to 6 min with a linear gradient from 2% solvent B to 3% solvent B, then isocratically from 3 to 18 min using 3% solvent B, 18 to 35 min with a linear gradient from 3% solvent B to 40% solvent B, 35 to 60 min using a linear gradient from 40 to 55% solvent B, then isocratically from 60 to 65 min at 55% solvent B then from 65 to 80 min from 55 to 0% B (linear) followed by equilibration from 80 to 90 min at 0% B

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