INTRODUCTION 1-Naphthylacetic and 2-naphthoxyacetic acids belong to the synthetic branch of auxins. Auxins have attracted considerable interest as a subject of study by virtue of their biological and physiological significance. Their broad use as plant growth regulators has raised the need for simple, rapid, sensitive and selective analytical methods for their determination in real samples. OBJECTIVE The primary aim of this work was to develop an analytical method for the simultaneous determination of 1-naphthylacetic acid and 2-naphthoxyacetic acid in commercial technical formulations, tomato and various fruit types (apple, strawberry, orange and plum) by room temperature phosphorescence. METHODOLOGY Filtrated solutions of aqueous slurries from ecological fruit and tomato samples are acidified and then extracted with dichloromethane. Once the solvent is evaporated, the dried residue is dissolved in sodium dodecyl sulphate (a micellar agent), and supplied with thallium (I) nitrate as an external heavy atom source and sodium sulphite as deoxygenation agent to enhance the ensuing phosphorescence. RESULTS The broad-band overlapping spectra for the two analytes were resolved by first- and second-derivative phosphorescence spectrometry. Zero-crossing measurements at 488.5 nm in the first-derivative spectrum and 469.5 nm in the second derivative spectrum exhibited a linear dependence on the 2-naphthoxyacetic acid and 1-naphthylacetic acid concentration, respectively. The detection limits as determined in accordance with the error propagation theory were 11.5 ng/mL for 1-naphthylacetic acid and 15.6 ng/mL for 2-naphthoxyacetic acid. CONCLUSION The proposed method affords the determination of 1-naphthylacetic acid and 2-naphthoxyacetic acid in real samples with near-quantitative recoveries from agricultural products.