The
separating the components is an important factor in the high-performance
liquid chromatography, for which the stationary and the mobile phase are used.
The choice of proper buffer for chromatographic analysis of analytes depends on
the peak value, pH range, and the characteristics of the buffer for UV cutoff.
The pH value of the buffer solution or mobile phase used within +/- 1 molecules
pKa value.
The
buffer is usually required when analyzing ionizable molecules with RP-HPLC. For
compounds such as these, the mobile phase pH determines whether they are
present in the form of ionized or nonionized. This creates a disparity since
ionized species always elute first from the HPLC column in reversed-phase
chromatography. If the pH of the buffer or mobile phase changes, the result
will be inconsistent. Buffers are also sometimes required for applications
because impurities or interfering compounds are ionizing. If the impurities or
interfering compounds are ionizable, then the buffers are also used.
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