When to use a gradient in HPLC? 

How do you know when you should use an gradient elution instead of isocratic elution?
In this exploration of gradients in chromatography, we unpack the pivotal question: when is a gradient necessary?
The heart of the matter lies in the hydrophobic similarities of the molecules being analyzed. If they share similar hydrophobicities, a gradient might not be needed, as indicated by a scouting run where peaks emerge closely together.
However, for a diverse range of hydrophobicities or unknown samples, gradients shine by offering versatility and the invaluable ability to clean the column after each run, effectively providing a 'new' column every time.
This piece demystifies the choice between gradient and isocratic methods, highlighting that while isocratic runs can be advantageous for their simplicity and efficiency in certain conditions, gradients are the go-to for their adaptability and comprehensive cleansing capability.
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