S-nitrosylation is an important post-translational modification which occurs on cysteine in organisms. S-nitrosylation regu-lates signal transduction in many cell processes and aberrant S-nitrosylated (SNO-) proteins are closely related to cardiovas-cular and neurological diseases. However, quantitative analysis of the S-nitrosylation at the proteome level remains challeng-ing due to the low abundance of the modification. Herein, we developed a novel approach called FluoroTRAQ, which com-bined the fluorous solid-phase extraction of SNO-peptides and the iTRAQ labeling for the quantitative analysis of SNO-proteome with high sensitivity and specificity. The new analytical strategy is successfully applied to monitor of the dynamic changes of SNO-proteome in cells. A quantitative dataset of SNO-proteome in human umbilical vein endothelial cell (HUVEC) after in vivo S-nitrosoglutathione (GSNO) stimulation was revealed. Among them, several new SNO-proteins in-cluding DHX9, GOT2 et al. are reported and validated by biotin switch assay.