In this work, carbon nanospheres (CNSs) were prepared with glucose through a hydrothermal synthesis method. Subsequently, a cetyltrimethylammonium bromide (CTAB) bilayer was used to modify the surface of the as-prepared CNSs (denoted as CTAB-CNSs). Interestingly, it was found that CTAB-CNSs could significantly amplify chemiluminescence (CL) emissions from some reactive oxygen species systems, including the Co(II)-H 2 O 2 -OH À system, ClO À system and ONOO À system. The CL intensity was proportional to the concentration of H 2 O 2 in the Co(II)-H 2 O 2 -OH À system in a linear range of 5-1000 mM with a limit of detection of 2.6 mM (S/N ¼ 3). The enhancement mechanism of CTAB-CNSamplified CL from the Co(II)-H 2 O 2 -OH À system was investigated using the fluorescence spectra, CL spectrum, scavengers of reactive oxygen species, and UV-vis absorption spectra. The results indicated that the CTAB bilayer on the surface of CNSs could form a particular micelle microenvironment, facilitating the reaction between reactive intermediates and CNSs. Finally, this method has been successfully used to detect H 2 O 2 in river water and lake water samples, and the recoveries were from 96 to 110%. The relative standard deviation (RSD) for nine repeated measurements was less than 5%. 010 64411957; Tel: +86 010 64411957 † Electronic supplementary information (ESI) available: Effect of the preparation time of CNSs on the CL intensity; TEM image of CNSs; FTIR spectrum of the as-prepared CTAB-CNSs and zeta potential measurements of CTAB-CNSs, CTAB and CNSs; effect of NaOH on the CL intensity; effect of CNSs modied with CTAB, TTAB, SDS and Triton X-100 on CL intensity; CL intensity of the ClO À -H 2 O 2 system and the ONOO À system in the presence of CTAB-CNSs; absorbance spectra of NBT in the absence or presence of the CTAB-CNS-Co(II)-H 2 O 2 -OH À system; tolerance limit of various coexisting substances on the determination of 50 mM H 2 O 2 ; effects of radical scavengers on the CTAB-CNS-Co(II)-H 2 O 2 -OH À system. See