Valorisation of the green waste parts from large-leaved buttercup (Ranunculus macrophyllus Desf.): phenolic profile and health promoting effects study

Due to the extensive use of Ranunculus macrophyllus Desf. roots for medicinal purposes, most of the leafy green parts are just wasted. The aim of this work is to valorize the leafy green parts and promote their application in different modern industries. Methods For this purpose, we studied the phenolic profile of R. macrophyllus Desf. (RM-B) using chromatographic and spectrophotometric methods and we tested the in-vitro antioxidant activity and the in-vivo effect of RM-B on plasma and liver antioxidant statuts. Results RM-B contained high amounts of polyphenols (675 mg GAE/100 g dry weigh dw) and flavonoids (105 mg QE/100 g dw). In-vitro, RM-B exhibited promising radical scavenging activity against 2,2′-azino-bis(3-éthylbenzothiazoline-6-sulphonique) (ABTS+·) (IC50: 247 µg/mL), hydrogen peroxide radicals (IC50: 626 µg/mL) and inhibited oxidative red blood cells hemolysis (IC50: 120 µg/mL), RM-B also showed strong reducing power (982 µM FeSO4/mg extract). In-vivo, RM-B improved the radical scavenging ability and reducing power of plasma and enhanced liver antioxidant status by increasing catalase and reduced glutathione levels and decreasing malondialdhyde levels without altering the key serum biochemical parameters reflecting liver and kidney functions. Polyphenols identified using capillary LC-DAD and LC–MS/MS analyses like hesperidin (131.2 mg/100 g dw), rutin (29.0 mg/100 g dw) and p-coumaric acid (5.8 mg/100 g dw), may be responsible for the health promoting effects of RM-B. Conclusion We may conclude that R. macrophyllus Desf. is a good source of beneficial polyphenols with strong antioxidant, anti-hemolytic and health-promoting effects, which promotes its use in pharmaceutical, medicinal and nutraceutical industries.Community of Madrid/ FEDER programMinistry of Economy and CompetitivenessDepto. de Química AnalíticaFac. de Ciencias QuímicasTRUEpu

Bioactive polyphenols from Ranunculus macrophyllus Desf. Roots: quantification, identification and antioxidant activity

Ranunculus macrophyllus Desf. is an Algerian medicinal plant whose roots are used in traditional medicine to cure feminine infertility and other diseases, however there are no studies regarding its phytochemistry and biological activities. The aim of this work is to study the phytochemical composition and antioxidant activity of different solvents fractions from the roots of Ranunculus macrophyllus Desf. Spectrophotometric and chromatographic methods were used to study the phytochemical composition; while antiradical, iron chelating ability, reducing power and lipid peroxidation were studied in-vitro. The ethyl acetate fraction showed the highest values of total phenolic compounds (271.0 ± 0.2 µg GAE/mg dry extract (d.e), flavonols (24 ± 5 µg RE/ mg d.e) and condensed tannins (129 ± 10 µg CE/mg d.e), while the hexane fraction contained the highest amount of triterpenoids (91 ± 7 µg UAE/mg d.e). The highest radical scavenging ability was recorded for the ethyl acetate fraction against DPPH (IC50 = 3.7 ± 0.1 µg/mL) and ABTS (IC50 = 81 ± 3 µg/mL) whereas the hexane fraction had the best hydrogen peroxide radical scavenging (IC50 = 380 ± 4 µg/mL). The ethyl acetate fraction had the best total antioxidant capacity (TAC = 361 ± 1 µAAE/ mg extract) and reducing power (310 ± 2 µAAE/mg extract). The β-carotene bleaching was inhibited at high rate even after 24 h by the ethyl acetate fraction (81.0 ± 0.5 %). All activities were correlated with the polyphenolic content of the fractions. Capillary LC-DAD and LC-MS/MS analysis of ethyl acetate fraction revealed high amounts of gallic acid (9.3 ± 0.6 mg/g d.e), dihydroxybenzoic acid (8.1 ± 0.2 mg/g d.e) and hesperidin (5.9 ± 0.6 mg/g d.e). With such high amounts of polyphenols and strong antioxidant activity Ranunculus macrophyllus Desf. roots could have a potential use in pharmaceutical and nutraceutical industries.Community of Madrid/FEDER programMinistry of Economy and CompetitivenessComplutense UniversityDepto. de Química AnalíticaFac. de Ciencias QuímicasTRUEpu

Anti-inflammatory activity of ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. and their phenolic profile

Ethnopharmacological relevance: The members of the genus Ranunculus have counter-irritating properties and thus, they are traditionally used for treating anti-inflammatory disorders and other skin conditions. Ranunculus macrophyllus Desf. is a wild medicinal plant growing in Algeria and traditionally used to treat some cutaneous skin disorders. Aim: The aim of this study was to characterize the composition of the ethyl acetate and n-butanol extracts from Ranunculus macrophyllus Desf. as well as to elucidate and to compare their effect against acute skin inflammation. Moreover, both the antioxidant activity and the acute toxicity of the plant extracts were also studied. Materials and methods: Spectrophotometric and chromatographic methods were employed to identify and quantify phenolic compounds and triterpenoids from R. macrophyllus Desf. fractions. The antioxidant activity was estimated using the phosphomolebdenum, DPPH, reducing power and β-carotene bleaching assays. The ethyl acetate and n-butanol extracts were screened for their anti-inflammatory activities using ex-vivo membrane stabilizing assays and in-vivo acute skin inflammation model. Results: Ethyl acetate fraction showed the highest amounts of total phenolic compounds (413 ± 4 μg GAE/mg extract) and triterpenoids (70.4 ± 1.8 μg UAE/mg extract). Rutin, hesperidin, myricetin and kaempferol were the major compounds identified in the different fractions. Ethyl acetate fraction exhibited strong DPPH• radical scavenging ability (IC50 1.6 ± 0.2 μg/mL), high total antioxidant capacity (447 ± 7 μg AAE/mg extract) and reducing power (514 ± 8 μg AAE/mg extract). Ethyl acetate fraction inhibited (73.4 ± 0.3) % of linoleic acid peroxidation. Ethyl acetate and n-butanol fractions did not have any visible toxicity at 2000 mg/kg and presented excellent membrane stabilizing ability. The inhibition of xylene induced ear inflammation was (38 ± 4) % and (46 ± 1) % for RM-B and RM-EA, respectively. Conclusions: The high content of both phenolic compounds and triterpenoids combined with the remarkable antiinflammatory effect and antioxidant activity of ethyl acetate and n-butanol extracts from R. macrophyllus Desf. support the wide spread use of this traditional plant on some skin disorders (inflammatory skin disorders).Community of Madrid/FEDER programMinistry of Economy and CompetitivenessDepto. de Química AnalíticaFac. de Ciencias QuímicasTRUEpu