Phytochemical analysis and biological activity of Prunus mahaleb L., P. spinosa L., Rubus discolor Weihe & Nees and R. serpens Weihe ex Lej & Court extracts (Rosaceae)

Циљ ове докторске дисертације је био испитивање хемијског састава и биолошких активности екстраката листова и плодова P. mahaleb, P. spinosa, R. discolor и R. serpens. Екстракти су добијени ултразвучном екстракцијом екстрагенсима различите поларности (вода, етанол и ацетон). Естракти листова и плодова R. discolor и R. serpens су били богатији фенолним једињењима од екстраката врста рода Prunus. LC-MS/MS методом је квантификовано 11 фенолних киселина и семиквантификовано 5 антоцијана. У њиховим екстрактима преовлађивале су хлорогенска, кафена и гална киселина, док је најзаступљенији антоцијанин у екстрактима плодова био цијанидин-глукозид/галактозид. Доминантне фенолне киселине у екстрактима P. mahaleb су биле p- и о-кумаринска, протокатехинска и гална, а код P. spinosa p-хидрокси-бензоева, гална, кафена, p-кумаринска и хлорогенска. Водени екстракт листа R. serpens је показао најјачу антиоксидантну активност у коришћеним тестовима (DPPH, ABTS, FRAP и TRC), осим у случају ß-каротен/линолеинска киселина теста где је највећу активност испољио ацетонски екстракт плода исте врсте. Антимикробна активност је одређена микродилуционом методом, а екстракти R. discolor и R. serpens су испољили јаче антибактеријско и антифунгално дејтво у односу на екстракте врстa рода Prunus. Цитотоксична активност различитих екстраката према HeLa, K562 и MDA-MB-453 ћелијским линијама је испитана MTT тестом. Показано је да екстракти листова R. discolor и R. serpens имају значајнији цитотоксични ефекат него екстракти врста рода Prunus. Морфолошком анализом ћелијске смрти и Annexin V-FITC/PI методом бојења је утврђено да екстракти цитотоксични ефекат остварују апоптозом малигних ћелија, а анализом дистрибуције ћелија по фазама ћелијског циклуса да доводе до пораста G1 ћелија, као и пораста броја апоптотичних (subG1 пик) ћелија. Сви испитивани екстракти су успешније инхибирали α-глукозидазу, него α-амилазу, при чему су се посебно истакли етанолни екстракти листова R. discolor и R. serpens. У овом раду су први пут представљени резултати хемијског састава и биолошких активности за врсту R. serpens, као и антидијабетичне активности за екстракте листова преостале три врсте. Добијени резултати указују да екстракти листова и плодова испитиваних врста испољавају бројне биолошке активности пре свега захваљујући синергистичком дејству фенолних једињења, те да се потенцијално могу применити као нутрацеутици и у фитофармацији.This study was aimed to evaluate the chemical composition and bioactivity of P. mahaleb, P. spinosa, R. discolor and R. serpens leaves and fruits. The extracts of the above-mentioned species were obtained by ultrasound-assisted extraction using solvents of different polarity (water, ethanol and acetone). The extracts of R. discolor and R. serpens species contained higher amounts of phenols than extracts of Prunus species. LC-MS/MS analysis enabled the identification of 11 phenolic acids and semi-quantification of 5 anthocyanins. The dominant phenolic acids in R. discolor and R. serpens extracts were chlorogenic, caffeic and gallic, whilst cyanidin-glucoside/galactoside was the main anthocyanin in their fruit extracts. The main phenolic acids in P. mahaleb extracts were p- and о-coumaric, protocatechuic and gallic and in P. spinosa p-hydroxybenzoic, gallic, caffeic, p-coumaric and chlorogenic acids. The aqueous leaf extract of R. serpens exhibited the highest antioxidant activity in all used tests (DPPH, ABTS, FRAP and TRC tests), except in ß-caroten-bleaching assay where its acetone extract was the most active. Antimicrobial activity was determined by microdilution protocol and R. discolor and R. serpens showed stronger antibacterial and antifungal activity than examined Prunus species. The cytotoxic activity towards HeLa, K562 and MDA-MB-453 human malignant cell lines was evaluated by MTT test. The leaf extracts of R. discolor and R. serpens possessed more significant cytotoxic effects than extracts of Prunus species. Morphological analysis of cell death and Annexin V-FITC/PI method proved proapoptotic effects of extracts which were responsible for its cytotoxicity. The cell cycle distribution analyses showed that extracts increased number of G1 cells as well as number of apoptotic cells (subG1 peak). All examined extracts showed better α-glucosidase than α-amylase inhibitory effects whereby R. discolor and R. serpens ethanol leaf extracts were particularly active. In this study, results of chemical composition and biological activity of R. serpens, as well as results of antidiabetic effects of leaf extracts of other three species, were presented for the first time. The presented results indicate that the examined plant species exhibited promising bioactivities mostly due to synergism of present phenolic compounds and therefore could be potentially applied as nutraceuticals and in phytopharmacy

Examination of the polyphenol content and bioactivities of Prunus spinosa L. fruit extracts

We investigated the total phenolic and flavonoid contents and the anthocyanin profiles in aqueous, ethanol and acetone extracts of Prunus spinosa (Rosaceae) fruit, and their antioxidant, antibacterial, antifungal, antidiabetic and antitumor properties. The contribution of polyphenol contents to the bioactivity of the extracts was calculated and observed through Pearson’s coefficient of correlation. The acetone extract was the richest in phenols and anthocyanins and the ethanol extract in flavonoids. Cyanidin was the most abundant anthocyanin compound in all examined extracts. The ethanol extract showed the most promising antioxidant activity in DPPH, ABTS and FRAP assays. Tested bacteria were more affected by the ethanol than by the aqueous extract. Both the ethanol and aqueous extracts exhibited potential antidiabetic effects, observed as inhibition of α-amylase and α-glucosidase, enzymes linked with diabetes mellitus type II. The ethanol extract was a potent α-glucosidase-inhibitor with a significantly lower IC50 value than the positive control, glucobay, used to treat diabetes mellitus type II. Neither the ethanol nor the aqueous extracts had any effects on tested human malignant cell lines. Our results indicate that the ethanol extract showed the most pronounced in vitro antioxidant and antimicrobial effects, and a potential antidiabetic activity, which can be ascribed to its high flavonoid content. Our results indicate that research of compounds, particularly of flavonoids present in the ethanol extract and their anti-diabetic properties should be examined further

Bioactivity and phenolics profile of aqueous and ethyl acetate extracts of Satureja kitaibelii Wierzb. ex Heuff. obtained by ultrasound-assisted extraction

The aim of the study was to investigate the biological activity and chemical composition of Satureja kitaibelii Wierzb. ex Heuff. LC–PDA/MS analyses for the aqueous extracts (A1-stem, leaves and flowers, A2-leaves and flowers) and ethyl-acetate extracts (E1-stem, leaves and flowers, E2-leaves and flowers) obtained by ultrasound-assisted extraction enabled the identification of thirty-four compounds. Quantitative analysis revealed that the aqueous extract obtained from leaves and flowers was the richest in total phenolic acids (65.36 mg/g) and flavonoids (21.17 mg/g). The total polyphenol content was the highest in the aqueous extract obtained from leaves and flowers (274 ± 2.4 mg Gallic Acid equivalents/g). The best antioxidant activity was observed for the same extract using the DPPH (SC50 20 ± 10 µg/mL), ABTS (2.834 ± 0.02 mg Ascorbic Acid/g), FRAP (1.922 ± 0.03 mmol Fe2+/mg), and total reducing power tests (16.4 ± 1.0 mg Ascorbic Acid/g). Both ethyl acetate extracts were the most active against strains of Bacillus cereus and Micrococcus flavus (MIC 1.70–1.99 mg/mL and 1.99–3.41 mg/mL, respectively). They were more efficient against Aspergillus ochraceus (MFC 0.86 mg/mL) and towards HeLa cell lines. All the obtained results implied the good potential of the investigated extracts to be used as effective preservatives and functional ingredients in food products and dietary supplements

The Effects of Sunflower and Maize Crop Residue Extracts as a New Ingredient on the Quality Properties of Pork Liver Pâtés

The present study aimed to evaluate the antioxidant capacity of ethanolic extracts from post-harvest sunflower and maize stalk residues, and their impact on the chemical composition, physicochemical parameters, lipid oxidative stability, microbiological properties, and sensory characteristics of pork liver pâtés over a 90-day storage period. Four formulations were prepared: a control group (CON), a batch with butylated hydroxytoluene as a synthetic antioxidant (BHT), 1% ethanolic extract from sunflower residues (SSRE), and 1% ethanolic extract from maize residues (MSRE). The MSRE had a higher total phenol content and showed better antioxidant activity relative to the SSRE (p < 0.01). The addition of SSRE decreased the lightness and increased the redness in the pork liver pâtés, with these pâtés showing the highest total color difference compared to the control (p < 0.01). The crop extracts increased the n-6 and total PUFA contents in pâtés and improved the PUFA/SFA ratio (p < 0.01). Formulations containing crop residue extracts showed higher TBARs and POV values than the control and BHT group (p < 0.01), indicating a pro-oxidant effect and accelerated lipid oxidation in pâtés during storage. As far as microbiological quality, the presence of crop residue extracts decreased the total viable count, lactic acid bacteria, and psychotropic aerobic bacteria (p < 0.01). The incorporation of crop extracts in the pork pâtés impaired their sensory quality, particularly color, odor, aroma, and flavor, and decreased their overall acceptability. These results indicated that, while the crop residue extracts were not as effective as synthetic antioxidants in preserving the lipid stability of pâtés, they demonstrated potential for enhancing the microbial quality of this type of meat product

Phytochemical analysis and biological activity of Prunus mahaleb L., P. spinosa L., Rubus discolor Weihe & Nees and R. serpens Weihe ex Lej & Court extracts (Rosaceae)

Циљ ове докторске дисертације је био испитивање хемијског састава и биолошких активности екстраката листова и плодова P. mahaleb, P. spinosa, R. discolor и R. serpens. Екстракти су добијени ултразвучном екстракцијом екстрагенсима различите поларности (вода, етанол и ацетон). Естракти листова и плодова R. discolor и R. serpens су били богатији фенолним једињењима од екстраката врста рода Prunus. LC-MS/MS методом је квантификовано 11 фенолних киселина и семиквантификовано 5 антоцијана. У њиховим екстрактима преовлађивале су хлорогенска, кафена и гална киселина, док је најзаступљенији антоцијанин у екстрактима плодова био цијанидин-глукозид/галактозид. Доминантне фенолне киселине у екстрактима P. mahaleb су биле p- и о-кумаринска, протокатехинска и гална, а код P. spinosa p-хидрокси-бензоева, гална, кафена, p-кумаринска и хлорогенска. Водени екстракт листа R. serpens је показао најјачу антиоксидантну активност у коришћеним тестовима (DPPH, ABTS, FRAP и TRC), осим у случају ß-каротен/линолеинска киселина теста где је највећу активност испољио ацетонски екстракт плода исте врсте. Антимикробна активност је одређена микродилуционом методом, а екстракти R. discolor и R. serpens су испољили јаче антибактеријско и антифунгално дејтво у односу на екстракте врстa рода Prunus. Цитотоксична активност различитих екстраката према HeLa, K562 и MDA-MB-453 ћелијским линијама је испитана MTT тестом. Показано је да екстракти листова R. discolor и R. serpens имају значајнији цитотоксични ефекат него екстракти врста рода Prunus. Морфолошком анализом ћелијске смрти и Annexin V-FITC/PI методом бојења је утврђено да екстракти цитотоксични ефекат остварују апоптозом малигних ћелија, а анализом дистрибуције ћелија по фазама ћелијског циклуса да доводе до пораста G1 ћелија, као и пораста броја апоптотичних (subG1 пик) ћелија. Сви испитивани екстракти су успешније инхибирали α-глукозидазу, него α-амилазу, при чему су се посебно истакли етанолни екстракти листова R. discolor и R. serpens. У овом раду су први пут представљени резултати хемијског састава и биолошких активности за врсту R. serpens, као и антидијабетичне активности за екстракте листова преостале три врсте. Добијени резултати указују да екстракти листова и плодова испитиваних врста испољавају бројне биолошке активности пре свега захваљујући синергистичком дејству фенолних једињења, те да се потенцијално могу применити као нутрацеутици и у фитофармацији.This study was aimed to evaluate the chemical composition and bioactivity of P. mahaleb, P. spinosa, R. discolor and R. serpens leaves and fruits. The extracts of the above-mentioned species were obtained by ultrasound-assisted extraction using solvents of different polarity (water, ethanol and acetone). The extracts of R. discolor and R. serpens species contained higher amounts of phenols than extracts of Prunus species. LC-MS/MS analysis enabled the identification of 11 phenolic acids and semi-quantification of 5 anthocyanins. The dominant phenolic acids in R. discolor and R. serpens extracts were chlorogenic, caffeic and gallic, whilst cyanidin-glucoside/galactoside was the main anthocyanin in their fruit extracts. The main phenolic acids in P. mahaleb extracts were p- and о-coumaric, protocatechuic and gallic and in P. spinosa p-hydroxybenzoic, gallic, caffeic, p-coumaric and chlorogenic acids. The aqueous leaf extract of R. serpens exhibited the highest antioxidant activity in all used tests (DPPH, ABTS, FRAP and TRC tests), except in ß-caroten-bleaching assay where its acetone extract was the most active. Antimicrobial activity was determined by microdilution protocol and R. discolor and R. serpens showed stronger antibacterial and antifungal activity than examined Prunus species. The cytotoxic activity towards HeLa, K562 and MDA-MB-453 human malignant cell lines was evaluated by MTT test. The leaf extracts of R. discolor and R. serpens possessed more significant cytotoxic effects than extracts of Prunus species. Morphological analysis of cell death and Annexin V-FITC/PI method proved proapoptotic effects of extracts which were responsible for its cytotoxicity. The cell cycle distribution analyses showed that extracts increased number of G1 cells as well as number of apoptotic cells (subG1 peak). All examined extracts showed better α-glucosidase than α-amylase inhibitory effects whereby R. discolor and R. serpens ethanol leaf extracts were particularly active. In this study, results of chemical composition and biological activity of R. serpens, as well as results of antidiabetic effects of leaf extracts of other three species, were presented for the first time. The presented results indicate that the examined plant species exhibited promising bioactivities mostly due to synergism of present phenolic compounds and therefore could be potentially applied as nutraceuticals and in phytopharmacy

Chemical Composition, Antioxidant, and Enzyme-Inhibitory Potential of Wheat Harvest Residue

Wheat (Triticum aestivum L., Poaceae), the most common cereal in the human diet in Europe and the United States, generates an enormous amount of underutilized agricultural byproducts. ...PSE Meeting 2023, 12-15 October 2023, Antalya, TürkiyeSaopštenje sa međunarodnog skupa štampano u izvodu (M34

Phytochemical Composition and In Vitro Bioactivity of Maize Harvest Residue

Maize (Zea mays L., Poaceae) is one of the most commonly grown cereals after wheat and rice. ...PSE Meeting 2023, 12-15 October 2023, Antalya, TürkiyeSaopštenje sa međunarodnog skupa štampano u izvodu (M34

Valorization of Sunflower Waste Ethanol Extract and Its Ethyl-Acetate Fraction

The sunflower (Helianthus annuus L.), which belongs to the Asteraceae family, is an annual plant native to North America. It is grown on approximately 28 million hectares worldwide, making it the third most important oilseed crop after soybean and rapeseed. ...PSE Meeting 2023, 12-15 October 2023, Antalya, TürkiyeSaopštenje sa međunarodnog skupa štampano u izvodu (M34

Could Agricultural Waste Material of Wheat, Maize and Sunfower Waste Be A New Source of Bioactive Compounds?

One of the most current problems is ensuring food supply for the growing population, which has led to a tripling of agricultural production in recent decades. ...PSE Meeting 2023, 12-15 October 2023, Antalya, TürkiyeSaopštenje sa međunarodnog skupa štampano u izvodu (M34

Antioxidant properties of Rubus discolor leaf extracts

In this work were examined aqueous, methanol, ethanol and acetone leaf extracts of Rubus discolor, wild growing blackberry, for their antioxidant properties and total phenol and flavonoid content. The total phenol content (TPC) varried from 250.05 to 446.61 mg GAE/g of dry extract, while total flavonoid content (TFC) was in range between 22.44 and 61.15 mg QE/g of dry extract. Aqueous extracts were the richest in phenols, as well as in flavonoids. In vitro antioxidant capacity of leaf extracts was evaluated by 2,2-diphenyl-1-picrylhydrazil (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical scavenging procedures and ferric reducing ability of plasma (FRAP) assay. Aqueous extracts were the most effective through all antioxidant tests. The total phenol content highly correlated with antioxidant activity of extracts. Moreover, weak correlation was established between total phenol and total flavonoid content. The results presented in this work indicate that phenol compounds contribute to antioxidant ability of extracts. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 173029