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Think of all you've already done

A Cupa Cupa Cupa Cupa Chemicals Caffeine has received a great deal of attention ever since it was identified as the principle stimulant in coffee (1820)

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The gradient started with 0.5% solvent B at a flow rate of 0.4 mL/min for 1 min and then ramped up to 95% solvent B at a flow rate of 0.35 mL/min over a period of 3 min