The characteristics of the assembling behavior of three disubstituted alkane derivatives, 1,16-hexadecanediol (HO(CH2)(16)OH), 1,18-octadecanedionic acid (HOOC(CH2)(18)COOH), and 16-hydroxyhexadecanoic acid (HO(CH2)(15)COOH) have been investigated by use of scanning tunneling microscopy (STM). Three different kinds of supramolecular structures on graphite surfaces are observed, suggesting that intermolecular hydrogen bonding between two carboxyl groups of HO(CH2)15COOH molecules is the preferential one among carboxyl and hydroxyl groups. The preference is also confirmed by the self-assembled structures of 16-mercaptohexadecanoic acid and the coadsorptions of HO(CH2)(16)OH with p-phthalic acid (PA) and HO(CH2)(15)COOH with PA. Theoretical simulation results on these systems support the STM observations.