To analyze the relationship between plasma apoB100 concentrations, apoB100/apoA-I ratio and the alteration of HDL subclasses distribution.The apoA-I contents of plasma HDL subclasses were quantitated by 2-dimensional gel electrophoresis coupled with immunodetection in 506 subjects.The subjects in the highest tertile of apoB100 groups had significantly higher small-sized preβ1–HDL, however lower large-sized HDL2a, HDL2b than the subjects in the lowest tertile of apoB100 group. Furthermore, a significant down in the HDL2b/preβ1–HDL values (from 6.8 to 1.6) with a rise in apoB100/apoA-I ratio (from 0.4 to 1.4) were observed. Compared to subjects with apoB100/apoA-I ratio < 0.9, the subjects with apoB100/apoA-I ratio ≥ 0.9 had significantly higher small-sized preβ1–HDL whereas lower HDL3a, HDL3b and large-sized HDL2a, HDL2b. Pearson correlation revealed that concentrations of apoB100 were positively correlated with preβ1–HDL but negatively correlated with HDL2a and HDL2b, and in multivariate analysis, all HDL subclasses were independently associated with the apoB100/apoA-I ratio.The apoB100 concentrations, especially apoB100/apoA-I ratio could reflect sensitively the alteration of HDL subclasses distribution. And HDL subclasses distribution characteristics of hyperlipidemic subjects appeared in the subjects with apoB100/apoA-I ratio ≥ 0.9, which indicated the efficiency of RCT was weakened and the maturation of HDL was blocked.