We flush our HPLC column with methanol followed by filtered deionised water and finally with mobile phase(ammonium acetate + glacial acetic acid + DI water) before we start running samples. Why is it necessary to flush with filtered DI water before flushing with mobile phase? Why can't we flush with methanol and go straight to flushing column with mobile phase?
(we run our samples using gradient elution- methanol for pump A and mobile phase for pump B)
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