Several species of Doryphorina leaf beetles from Central- and South America produce oleanane triterpene glycosides in their defensive glands. The presence of pentacyclic triterpenes in insects is intriguing since they lack the key enzymes necessary to synthesize these compounds. Since beta-amyrin is a common constituent of leaf waxes, we hypothesized that these leaf beetles use this compound as a precursor to their oleanane glycosides. To test this hypothesis we first confirmed the presence of beta-amyrin in Ipomoea batatas, the food plant of Platyphora kollari. Next, adults of P. kollari were fed for 10 days with I. batatas leaf disks painted with a solution of [2,2,3-(2)H(3)]beta-amyrin ([2,2,3-(2)H(3)]-1). The secretion from their defensive glands was collected and analyzed by HPLC-ESIMS. The results demonstrated that the secretion of beetles fed with an amount of [2,2,3-(2)H(3)]beta-amyrin corresponding to the quantity of unlabeled (natural) beta-amyrin present in the leaf disks contained on average 5.1% of [2,2,3-(2)H(3)]-3- O-beta- d-glucopyranosyl-(1-->4)-beta- d-glucuronopyranosyl-hederagenin ([2,2,3-(2)H(3)]-2), whereas the secretions of beetles fed with 10 times this amount of [2,2,3-(2)H(3)]beta-amyrin contained on average 23.9% of [2,2,3-(2)H(3)]-2. In both series of experiments, the percentage of labeled versus unlabeled triterpene glycoside in the secretion was positively correlated with the amount of deuterated beta-amyrin ingested. These results demonstrate for the first time that some leaf beetles are able to metabolize a widespread triterpenic constituent of leaf wax into more complex glycosides that are stored in their defensive glands.