High-performance liquid chromatography is a technique of separation using the stationary phase and mobile phase, in which buffer solutions are also sometimes used to separate ionizable compounds. An accurately buffered mobile phase is very significant in a reversal-phase chromatography (RP-HPLC) method for symmetrical peak shape, detection limits, and reliable retention times. If you know analytical functional groups and pKa values of the molecule then it is simple to choose the appropriate pH of buffer solution.
In RP-HPLC, the pH of the mobile phase value is usually between 02 and 08. Reversed-phase chromatography requires buffers when the component is ionizable, or if the pH of the sample solution is outside the pH range.
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