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Performance evaluation of HPLC column
Sep 14, 2017

In addition to the performance of the stationary phase, the Chromatographic column is also related to the filling technique. Under normal conditions, the dry filling preparation column is suitable when the filler grain size is >20um, and the wet filling is ideal when the granules are <20um. There are generally 4 kinds of filling methods: ① high-pressure homogenate method, which is used for the analysis of column and small-scale preparation column filling; ② radial pressure method, Waters patent, ③ axial compression method, mainly used for filling large diameter columns; ④ dry method. Column filling is highly technical, and most laboratories use the filled product column.

It must be pointed out that the acquisition of high performance liquid chromatographic column and the filling technology is an important link, but the fundamental problem lies in the merits and demerits of the filler itself, and whether the structure of the matching chromatograph system is reasonable.

Whether it is the filling of their own or the purchase of the column, before use to the performance of its investigation, use period or after a period of time to be checked again. Column performance index includes the column pressure under certain experimental conditions (sample, flow phase, velocity, temperature), theoretical tray height and tray number, symmetry factor, capacity factor and selectivity factor repeatability, or separation degree. Generally speaking, the repeatability of the capacity factor and the selectivity factor is within ±5% or ± 10%. When comparing the column effect, it is also necessary to pay attention to whether there is any change.

A Qualified chromatographic column evaluation report should give the basic parameters of column, such as column length, inner diameter, type of filler, particle size, column effect of chromatographic column, asymmetry and column pressure drop, etc.

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