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In 200 min of analysis, the flow rate was set to 0.25 L/min, and the linear gradient of buffer B was set as followed: 2% at 0 min, 2% at 5 min, 10% at 6 min, 30% at 135 min, 54% at 155 min, 95% at 157 min, 95% at 180 min, 95% at 180 min and 2% at 200 min

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