Three-factor two-level fractional experimental design, FFE 23-1 was used in order to evaluate the influence of Ethyl Alcohol (X1), sodium carbonate / citric acid, H3Cit (X2) and polygalacturonase enzyme, PGU (X3) to the extraction of biologically active compounds (BACs) from the septum. Septum samples, 2 g each, were extracted with 50mL of solution, prepared according to Table 1. Table 1. Extraction conditions planning matrix: encoded and real coordinatestableExtracted BAC per 100g of septum, mg/100g, of Gallic Acid (Gal), Catechin (Cat), Epicatechin (Epi), Casuarictin (Cas) and Ellagic Acid (Ell), were determined by reversed-phase HPLC with Photo Diode Array detection on C18-column. Following regressions were calculated: Gal = 8.4X0 – 3.0X1 – 2.8X2 + 3.6X3 Cat = 107X0 + 17X1 – 8X2 + 11X3 Epi = 27.5X0 – 7.5X1 – 14.0X2 + 7.8X3 Cas = 22.4X0 + 2.8X1 + 0.7X2 – 4.2X3 Ell = 32.6X0 + 2.2X1 + 4.8X2 – 6.6X3 The regressions show that the factors under research, have ambiguous effects on the extraction of biologically active compounds (BACs) from the septum. Thus, high concentrations of ethyl alcohol reduce the yield of Gal, but increase the extractivity of hexahydroxydiphenic acid derivatives (Cas and Ell). Alkaline medium (Na2CO3) reduce the yields of Gal, Cat and Epi, but increase the yields of Cas and Ell. Finally, the effects of polygalacturonase are also grouped according to BACs polarity: enzyme facilitating the release of more polar BACs. The totality of the factors influence can be explained by the interaction of BACs with different polarity with the biopolymer (cellulose-lignin) matrix of the septum. The most acceptable conditions for the extraction of all biologically active substances at once are close to the center of the experiment: ethanol-water mixture 30:70 containing H3Cit (~ 0.025%) and PGU (~ 0.005%).