In this paper,an application of microchip electrophoresis with chemiluminescence detection（MCE-CL） in single human red blood cell analysis was described.Human red blood cells were assayed to determine the intracellular content of hemoglobin（Hb）.The injection of individual cells,in-line lysed,and MCE separation were integrated onto a simple cross microfluidic chip.CL detection was based on the catalytic effects of hemoglobin on the luminol-hydrogen peroxide reaction.The MCE-CL detection had a linear calibration curve over the range of 2.0-90 pg injected with a correlation coefficient of 0.998 9 and a detection limit（S/N=3） of 0.8 pg.The content of Hb in individual human red blood cell was found in the range of 14-68 pg（n=19）.Compared to other methods,the MCE-CL method is simple and sensitive,and could be an attractive alternative for single cell analysis.