Authentication of holy basil using markers relating to a toxicology-relevant compound

Holy Basil—Ocimum tenuiflorum—is one of the popular new “superfoods” thought to act as an antioxidant and to reduce stress and anxiety. However, it is often surrogated with other Ocimum species differing in their chemical profiles that may even pose health risks to the consumers. Moreover, even specific chemotypes of Holy Basil itself can be toxicologically relevant, because they sometimes contain the carcinogen compound methyleugenol. Using DNA barcoding based on plastidic markers, O. tenuiflorum can be differentiated from other species of Ocimum. However, this approach is still suboptimal in handling larger sample numbers and in tracing chemotypes that accumulate methyleugenol. We have, therefore, designed a trait-related DNA barcode based on the enzyme eugenol O-methyltransferase (EOMT), responsible for the synthesis of methyleugenol. We show that a multiplex PCR combining trait-related and trait-independent markers can differentiate O. tenuiflorum from other Ocimum species and identify methyleugenol chemotypes of O. tenuiflorum, even in dried material sold as mixtures.