| In this paper, an ion-exchange/reversed phase liquid chromatography system (2D-LC) was developed based on two trap columns and a 10-port valve interface. The weak retained components of the first dimension were collected and analyzed separately, while the remained components were further separated by the second dimension of the 2D-LC system. The overloading of the second-dimensional column can be avoided effectively and the separation power of the 2D-LC system was improved greatly. The typical digest of tumor tissue, derivatived by 4-fluoro-7-nitro-2,1,3-benzoxadiazole(NBD-F), was separated and detected by 2D-LC-LIFD system,200 peaks more were detected compared to that measured by 2D-LC-UV system. The above result reveals that more efficient separation and higher sensitive detection of proteomes can be realized by the 2D-LC system coupled with LIFD.The molecular structures and the abundance of proteins studied in proteome are quite different, so it is necessary to use high-performance fluorescence reagents in LIF detection.In this paper,perylene,4-nitro-1,8-naphthalic anhydride and N-n-butyl-4-aminoethyl amido-1,8-dicarboximide were used as the fluorescence reagents to label compounds containing amino group. The results shows that 4-nitro-1,8-naphthalic anhydride can be employed to label amino acids and alkyl amine; while aldehydes can efficiently be derivated by N-n-butyl-4-aminoethyl amidonaphthalene-1,8-dicarboximide with a detection limit of 1.86×10-10 mol/L. This method enable the selective detection of glycoproteins and proteins containing aldehyde group be possible.
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