Porous graphitic carbon: A novel application as a first-dimension stationary phase in the 2D-LC-MS/MS analysis of complex peptide mixtures

摘要

From the experiments outlined above, we conclude that PGC performs well as a suitable off-line first dimension stationary phase in a 2D-LC-MS/MS proteomic study. Realistic biological quantities of starting material (approximately 400 (mu)g digested whole cell lysate) result in the identification of >13,000 peptides at an estimated FDR of 2percent using standard chromatographic equipment. Only 4 percent (approx 16 (mu)g) of this material was injected onto the second dimension. In addition, replicate analyses show good reproducibility and assessment of the column performance (100 consecutive gradient elutions) demonstrates the robust nature of the PGC material. We propose that PGC represents an excellent alternative stationary phase to both SCX and C18 phases operated under elevated pH conditions.