The optical rotatory dispersion spectra of echinulin and hydroechinulin have been compared with the spectra of the 4 stereoisomers of cyclo-alanyltryptophan. The results indicate appreciable differences in the spectra of the cis-and trans-piperazinediones and that echinulin has the cyclo-L-alanyl-L-tryptophan configuration.Canadian Journal of Chemistry, 47, 2069 (1969) Echinulin (la) is a white crystalline compound which was first isolated from a culture of Aspergillus echinulatus (Delacr) Thorn & Church (1). The structure (la) was established by a series of degradations and partial synthesis (2) and is supported by nuclear magnetic resonance (n.m.r.) (3) and biochemical (4) studies.Acid hydrolysis (5) of hexahydroechinulin (hydroechinulin) (lb) gave L-alanine and the amino acid hexahydroechinin (hydroechinin) However, echinin (2a) could not be obtained (5) by acid hydrolysis of echinulin (la) although hydroechinulin (lb) gave hydroechinin (2b) under the same conditions. The acid hydrolysis of hydroechinulin was previously used in this laboratory in connection with biosynthetic studies (7) but did not give a pure product. In view of this, and the observation (8) that the configuration of a piperazinedione can be determined by o.r.d., it was decided to compare the 0.r.d. spectra of echinulin and the model cyclo-alanyltryptophans (1, R, = R2 = H).The piperazinediones were prepared by the cyclization (9) of alanyltryptophan or tryptophylalanine as shown in Fig.