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Binary gradient elution protocol at a flow rate of 150 L/min using water/formic acid, 99.9/0.1 ( v/v ) (A) and acetonitrile/formic acid 99.9/0.1 ( v/v ) (B) varying between isocratic step for 1 min of 5% mobile phase B, then a linear increase of B from 5% to 100% over 11 min

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These small changes reduce the typical increase in bruising without changing the benefit of the shot