To deal with complicated separation situations, this study successfully prepared two mixed-mode chromatography (MMC) stationary phases, CCL-SIL and PCL-SIL, by functionalizing dialdehyde cellulose (DAC) derivatives. In liquid chromatography applications, CCL-SIL exhibited superior separation performance for nucleosides and bases in HILIC mode, while PCL-SIL performed better in RPLC and IEC modes. Their distinct separation mechanisms were also elucidated by quantum chemical calculations. Both CCL-SIL and PCL-SIL showed good stability and reproducibility, with relative standard deviations of retention time, peak area, and peak height below 7.79 % and 4.37 % for multiple injections. Particularly, the PCL-SIL column and the CCL-SIL column were successfully used for the quantitative analysis of trace targets in real samples with complex matrix, demonstrating high accuracy and precision.
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