The DNA interaction potency of a chemical has been defined in the present study as the degree of a chemical's ability to interact with DNA. An estimation method of such a potency has been established based on the peak reduction of an oligonucleotide probe resulting from its interaction with chemicals based on high-performance liquid chromatography. A DNA interaction potency equivalency (PEQ) also has been proposed to evaluate the relative interaction potency of test chemicals against benzo[a]pyrene-7, 8-dihydrodiol-9, 10-epoxide (BPDE). Five known direct DNA interaction chemicals were employed to demonstrate the method. Two known inactive chemicals were used as negative controls. Both the potency and PEQ(50) values (PEQ of testing chemical at 50% of the probe peak reduction) of these five chemicals were determined as BPDE > phenyl glycidyl ether (PGE) > tetrachlorohydroquinone (Cl4HQ) > methyl methanesulfonate (MMS) > styrene-7,8-oxide (SO). Among the reactive chemicals, MMS was found to break the oligonucleotide into smaller fragments, whereas BPDE, PGE, and SO form covalent adducts with the oligonucleotide. In the latter case, the formation of multi-chemical-oligonucleotide adducts also was observed by mass spectrometry. The method was employed to estimate the DNA interaction potency equivalency of diesel vehicle exhaust gas to demonstrate the applicability of this approach in evaluating the interaction potency of environmental pollutants in both gas and liquid phases.