Gradient HPLC serves as a crucial tool for analysing samples containing compounds with varying polarity. Its effectiveness stems from the ability to tailor both gradient time and composition, enabling the manipulation of analyte retention times. However, challenges arise from deviations in the programmed gradient profiles produced by HPLC systems, impacting retention time and peak shape. Several factors contribute to this variability in gradient HPLC, such as solvent quality, pump performance, changes in system pressure, and the condition of the columns. Addressing these challenges requires precise control and monitoring of gradient profiles.
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