| Abstract | Metodom odzivnih površina (RSM) provedena je optimizacija ekstrakcije potpomognute mikrovalovima (MAE), ubrzane ekstrakcije otapalima pri povišenom tlaku (ASE) i ekstrakcije potpomognute visokim hidrostatskim tlakom (HPAE) za dobivanje ekstrakta cvijeta trnine s najvišim udjelom ukupnih fenola, ukupnih flavonoida i ukupnih hidroksicimetnih kiselina. Kod sve tri tehnike ispitivan je utjecaj otapala (50 i 70 % vodene otopine etanola i metanola), zatim utjecaj temperature (40, 50 i 60 °C) i vremena ekstrakcije (5, 15 i 25 min) kod MAE, utjecaj temperature (60, 80 i 100 °C) i broja ciklusa ekstrakcije (1,2 i 3) kod ASE, utjecaj temperature (40 i 60 °C), vremena ekstrakcije (5 i 15 min) i tlaka (200 i 500 MPa) kod HPAE. Najviši maseni udjeli ukupnih fenola (69,80 mg GAE/g) određeni su u ekstraktima dobivenim pri optimalnim uvjetima MAE uz vodenu otopinu etanola. Najviši maseni udjeli ukupnih flavonoida (18,22 mg QE/g) i ukupnih hidroksicimetnih kiselina (24,81 mg CAE/g) određeni su u ekstraktima dobivenim pri optimalnim uvjetima ASE uz vodenu otopinu etanola. HPLC analizom je kao najdominantniji spoj određen kamferol-pentozid (19,87 mg/g u vodenoj otopini metanola, 18,55 mg/g u vodenoj otopini etanola) u ekstraktima dobivenim pri optimalnim uvjetima MAE, odnosno procijandin B1 (26,98 mg/g u vodenoj otopini metanola, 22,9 mg/g u vodenoj otopini etanola) u ekstraktima dobivenim pri optimalnim uvjetima ASE. U svim ekstraktima primjenom DPPH i FRAP metode određene su visoke vrijednosti antioksidacijskog kapaciteta, a najviše su određene u ekstraktima dobivenim pomoću HPAE i ASE. Primjenom in vitro metoda Kenacid Blue, Neutral Red i Trypan Blue utvrđen je antiproliferativan učinak cvijeta trnine na Hep G2 stanice, ovisan o koncentraciji ekstrakta i duljini inkubacije. Pri tretmanu stanica s najnižom koncentracijom ekstrakta cvijeta trnine (10 μg/mL) određen je najveći udio apoptotičkih stanica u kulturi (36,75 %). |
| Abstract (english) | Response surface methodology (RSM) was used for optimization of microwave assisted extraction (MAE), accelerated solvent extraction (ASE), high pressure-assisted extraction (HPAE) in order to obtain blackthorn flower extract with highest content of total phenols, total flavonoids and total hydroxycinnamic acids. In all three techniques, the effect of solvents was examined (50 and 70 % aqueous solutions of ethanol and methanol), then the effect of temperature (40, 50 and 60 °C) and time of extraction (5, 15 and 25 min) in MAE, effect of temperature (60, 80 and 100 °C) and number of cycles (1,2 and 3) in ASE, effect of temperature (40 and 60 °C), extraction time (5 and 15 min) and pressure (200 and 500 MPa) in HPAE. The highest content of total phenolic (69,80 mg GAE/g) was determined in extracts obtained under optimal MAE conditions with aqueous ethanol solution. The highest content of total flavonoids (18,22 mg QE/g) and total hydroxycinnamic acids (24,81 mg CAE/g) were determined in extracts obtained under optimal ASE conditions with aqueous ethanol solution. Using HPLC kaempferol pentoside was determined as the most dominant compound (19,87 mg/g in aqueous methanol solution, 18,55 mg/g in aqueous ethanol solution) in extracts obtained under optimal MAE conditions, ie procyanidin B1 (26,98 mg/g in aqueous methanol, 22,9 mg/g in aqueous ethanol) in extracts obtained under optimal ASE conditions. By using DPPH and FRAP methods high values of antioxidant capacity were determined in all extracts, and the highest were determined in extracts obtained by using HPAE and ASE. Using in vitro methods Kenacid Blue, Neutral Red and Trypan Blue, the antiproliferative effect of blackthorn flower on Hep G2 cells, depending on the extract concentration and incubation length, was determined. When treating the cells with the lowest concentration of blackthorn flower extract (10 μg/mL), the highest proportion of apoptotic cells in the culture was determined (36,75 %). |
