Epichlorohydrin (ECH) and tetraethylenepentamine (TEPA) were copolymerized and the copolymers were modified hydrophobically using a pendant C18 hydrophobic group (HPTE). Hydrophobic interaction of modified polymer was investigated by fluorescence using pyrene as a probe. It reveals that hydrophobic microdomains are formed by aggregation of hydrophobic side chains. The water in hydrophobic microdomains is in a state of diffusion equilibrium under the hydrophobic interaction. Critical aggregate concentration (c(cac)*) of HPTE decreases with increasing content of the bound hydrophobic groups.