HPLC-DAD analysis of statistical mixture design extracts of Erythrina speciosa Andrews leaves provided chromatographic and UV–visible profiles of their basic and organic fractions that were treated with the PARAFAC multivariate method. The design extracts provided greater varieties and amounts of metabolites than could be obtained by classical extraction methods. Fractionation provided more diverse fingerprint information than obtained previously from only the crude extract. The two largest chromatographic peaks, one with a 4.8 min elution time having an intense spectral band at 235 nm and the other a 5.8 min peak with an intense 238 nm band for the basic fraction were obtained with the ternary 1:1:1 ethanol–dichloromethane–hexane mixture. These can be assigned to diene-type and lactonic alkaloids. Peaks with the same retention times are also found in the organic fraction but are extracted with different mixtures and have distinct spectral behavior in the 235 nm region, probably being aromatic alkaloids. The above strategy permits a more unambiguous assignment of metabolic groups to specific chromatographic peaks. This can be expected to provide higher quality chromatographic fingerprints for natural products’ chemistry.