Friday, November 15, 2019

Why use a buffer in the HPLC mobile phase


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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