Friday, November 22, 2019

Why do we need buffers solution in HPLC?

When ionizable molecules are analyzed with inverted phase chromatography, the buffers usually need to separate them. The buffer resists the change in pH so that compounds and silica (column) will be ionized consistently, resulting in reproducible results of chromatography. A buffered mobile phase is selected to maintain consistent retention time (RT) and selectivity to separating acids and bases. Buffer salts effectively decrease peak tailing by masking silanols for basic analytes. They also decrease the interactions of potential ion exchange with unprotonated silanols. To separate the most basic compounds in reversed phase-HPLC, it is recommended that the buffer concentration should be in the range of 10 - 50 mM.



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