How Is The Performance of The HPLC Column?
In addition to the performance of the stationary phase, the performance of the HPLC column is also related to the filling technique. Under normal conditions, when the particle size of the filler is >20μm, the dry-filled column is suitable; when the particle is <20μm, the wet filling is ideal. There are generally four filling methods:
1. High-pressure homogenization method, mostly used for analysis column and small-scale preparation column filling;
2. Radial pressure method, Waters patent;
3. Axial pressure method, mainly used for filling large-diameter columns;
4. Dry methods. Column packing is highly technical and most laboratories use filled commercial columns.
It must be pointed out that the acquisition of high performance liquid chromatography column, loading technology is an important link, but the fundamental problem is the performance of the filler itself, and the structure of the matching chromatograph system is reasonable.
Whether you are filling your own or purchasing an HPLC column, check its performance before use, or recheck it during use or after a period of time. Column performance indicators include column pressure, theoretical plate height and plate number, symmetry factor, capacity factor and selectivity factor repeatability, or resolution under certain experimental conditions (sample, mobile phase, flow rate, temperature). In general, the repeatability of the capacity factor and the selectivity factor is within ±5% or ±10%. When comparing the efficiency of the column, pay attention to whether the extra-column effect changes.