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The influence of the factors “vegetation period” and “age of wood” over the physico-chemical properties of the walnut oils has been investigated and evaluated. The content of the β-carotene and chlorophyll, color parameters, refractive index and fluorescence properties of the investigated butters have been studied. The effects of both factors and the influence over the main characteristics of the walnut oil have been estimated by using the two-factor analysis of variance (ANOVA). The statistical significance of the evaluation and the strength of the effect of the factors have been determined by Snedekor method applying the F-criterion of Fisher.