Oxtotitlán Cave paintings have been considered among the earliest in Mesoamerica on stylistic grounds, but confirmation of this hypothesis through absolute dating has not been attempted until now. We describe the application of advanced radiocarbon strategies developed for situations such as caves with high carbon backgrounds. Using a low-temperature plasma oxidation system, we dated both the ancient paint and the biogenic rock coatings that cover the paint layers at Oxtotitlán. Our research has significantly expanded the time frame for the production of polychrome rock paintings encompassing the Early Formative and Late Formative/Early Classic periods, statistically spanning a long era from before ca. 1500 cal B.C. to cal A.D. 600.