One of the most common peak problems in HPLC is peak splitting or broadening. This can be caused by a variety of factors, (1).column overload,
(2).poor column packing.
In some cases, peak splitting can also be caused by sample matrix effects or poor sample preparation techniques. To overcome this issue, it is important to ensure that the HPLC system is properly maintained, and that the column is properly packed and equilibrated before each analysis.
Column Overload
Column overload happens when too much sample or solute is injected onto the column. Column overload reduces the performance of the column by causing retention time and peak shape problems.
Types of Over Load
There are two main types of column overloads.
(I). Mass Overload.
(II). Volume Overload.
Poor Column Packing
A poorly packed column gives rise to poor and uneven flow, band broadening, and loss of resolution.