The protective HPLC columns influence the separation result. First of all, the protective column has a good positive effect on the experiments. Even if the stationary phase is chosen incorrectly, the protective column can also be effective for separation. The protective column is used to protect the analytical HPLC columns from contamination by highly residual components. The protective columns are much cheaper than analytical ones, so contaminated ones can be replaced more frequently.
It’s best that the stationary phase of the protective HPLC columns is exactly the same as that of the analytical column. If the stationary phase has a high reservation, the separation will be affected due to easily retained in the columns. On the contrary, the stationary phase with a low reservation will not affect the separation obviously.
And in order to minimize the separation impact of the protective HPLC columns, the protective columns need to be properly assembled. They are inserted between the sample inlet and the analysis columns. If the pipeline is too long or has a big inside diameter, it will cause additional peak broadening of the result.
In theory, if the protective column lengthens the HPLC columns length, there will be an additional number of trays for chromatographic separation. The advantage of the protective column is to prolong the life of the column instead of improving the overall analysis efficiency.
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