Amphipyrinae have long been a catchall taxon for Noctuidae, with most members lacking discernible morphological synapomorphies that would allow their assignment to one of the many readily diagnosable noctuid subfamilies. Here data from seven gene regions (>5,500 base pairs) for more than 120 noctuid genera are used to infer a phylogeny for Amphipyrinae and related subfamilies. Sequence data for 57 amphipyrine genera—most represented by the type species of the genus—are examined. Presented here are: the first large-scale molecular phylogenetic study of Amphipyrinae and largest molecular phylogeny of Noctuidae to date; several proposed nomenclatural changes for well supported results; and the identification of areas of noctuid phylogeny where greater taxon sampling and/or genomic-scale data are needed. Adult and larval morphology, along with life history traits, for taxonomic groupings most relevant to the results are discussed. Amphipyrinae are significantly redefined; many former amphipyrines, excluded as a result of these analyses, are reassigned to other noctuid subfamily-level taxa. Four genera, Chamaeclea Grote, Heminocloa Barnes & Benjamin, Hemioslaria Barnes & Benjamin, and Thurberiphaga Dyar are transferred to the tribe Chamaecleini Keegan & Wagner New Tribe in Acontiinae. Stiriina is elevated to Stiriinae Revised Status, Grotellina is elevated to Grotellinae Revised Status, and Annaphilina is elevated to Annaphilini Revised Status. Acopa Harvey is transferred to Bryophilinae, Aleptina Dyar is transferred to Condicinae, Leucocnemis Hampson and Oxycnemis gracillinea (Grote) are transferred to Oncocnemidinae, Nacopa Barnes & Benjamin is transferred to Noctuinae, and Narthecophora Smith is transferred to Stiriinae. Azenia Grote (and its subtribe Azeniina), Cropia Walker, Metaponpneumata Möschler, Sexserrata Barnes & Benjamin, and Tristyla Smith are transferred to Noctuidae incertae sedis. Hemigrotella Barnes & McDunnough (formerly in subtribe Grotellina) is retained in Amphipyrinae.