This paper demonstrated the chemical analysis of single cell on a cross PDMS microfluidic chip in a simple, fast, and high-throughput mode. The pre-stained cells were sequentially loaded into the cross section by hydrodynamic force, lysed by 0.2% SDS and subsequently the lysates were detected by LIF. Each cell can be lysed within 500 ms due to its high concentration of SDS at cross section resulted from the absence of electro-osmosis after surface coating in microchannel. The reliability and quality of the analysis was confirmed by analysis of glutathione and rhodamine 123 in single K562 cells. In each run, approximately 100 cells could be analyzed in about 10 min, which demonstrated the comparatively high throughput. The proposed microfluidic method is simple, fast, and high throughput, which might be of significance in identifying the biological molecules involved in fast biochemical processes and studying heterogenous cells.