Dichloromethane extracts of dried flowers of Calendula officinalis contain eight known bioactive triterpendiol monoesters, namely, faradiol-3-O-palmitate, faradiol-3-O-myristate, faradiol-3-O-laurate, arnidiol-3-O-palmitate, arnidiol-3-O-myristate, arnidiol-3-O-laurate, calenduladiol-3-O-palmitate and calenduladiol-3-O-myristate. These pentacyclic terpenoids have been quantified simultaneously using reversed-phase HPLC with isocratic elution and internal standardisation. Of the 10 varieties of C. officinalis investigated, Calypso Orange Florensis produced the highest amounts of the bioactive monoesters, followed by Fiesta Gitana Gelb and May Orange Florensis. The lipophilic extract from the flowers of Calypso Orange Florensis variety also contained low levels of the newly characterised calenduladiol-3-O-laurate.