hrvatski

Sadržaj polifenola i antioksidacijska aktivnost imele (Viscum album L.) sakupljenih sa različitih domadara

Cilj ovog rada je ispitati sadržaj polifenola i antioksidacijsku aktivnost metanolnih ekstrakata (80%) bijele imele sakupljene u travnju 2017. na Lašćini i na području Maksimira u Zagrebu (Hrvatska) sa nekoliko različitih domadara (lipa - Tilia platyphyllos Scop. , glog - Crataegus monogyna Jacq. i javor - Acer campestre L). Tekućinskom kromatografijom visoke djelotvornosti s obrnutim fazama (RP-HPLC) detektirani su pojedinačni polifenoli, a količina ukupnih fenolnih spojeva određena je pomoću Folin- Ciocalteu metode. Tri različite metode korištene su za određivanje antioksidacijske aktivnosti ABTS (2, 2 –azinobis (3- ethylbenzothiazoline-6-sulfonic acid), DPPH (1, 1-diphenyl-2-picrylhydrazyl) i FRAP (Ferric Reducing/Antioxidant Power Assay). Dio listova imele sa peteljkama je liofiliziran dok je drugi dio sušen u sjeni i na sobnoj temperaturi. Osušeni biljni materijal usitnjen je u komadiće promjera približno 5 mm i ekstrahiran u 80 % etanolu u staklenoj koloni dimenzije 1×10 cm u trajanju od 24h. Perkolat je ispušten iz kolone brzinom od oko 4 kapi u minuti te uparen na rotavaporu na temperaturi od 30 °C . Metanol (80 %) je dodan kako bi se dobio omjer suhe biljne mase i otapala 1:1. HPLC analizom uzoraka listova imele detektirano je ukupno 8-10 fenolnih spojeva, od toga 5-7 fenolnih kiselina (p-kumarinska, galna, klorogenska, kavena, ferulična, sinapilna i salicilna kiselina u uzorcima imele koja je rasla na glogu, klorogenska kiselina nije detektirana u uzorcima imele koja je rasla na javoru i lipi, a galna kiselina nije detektirana u imeli koja je rasla na lipi) i 3 flavonoida (kvercetin, kempferol i isoramentin). Fenolne kiseline p- kumarinska i sinapilna te antioksidacijska aktivnost mjerena sa ABTS i DPPH metodom statistički značajno je bili viša u skoro svim liofiliziranim uzorcima u odnosu na uzorke imele sušene na sobnoj temperaturi. Najviša koncentracija ukupnih i individualnih fenola i generalno najviša antioksidacijska aktivnost detektirana je u uzorcima imele koja je rasla na glogu. Većina uzoraka imele imala je jaku antioksidacijsku aktivnost (>70%) u odnosu na pozitivnu kontrolu Trolox. Na temelju rezultata možemo zaključiti da uzorci imele koji su rasli na glogu predstavljaju najbolji izvor polifenola i antioksidanta koji se mogu primijeniti u fitoterapiji za liječenje različitih oboljenja. Također, liofilizacija ima pozitivan učinak na antioksidacijsku aktivnost (ABTS i DPPH) te na očuvanje koncentracije p-kumarinske i sinapilne kiseline u uzorcima imele./The aim of this study is investigated polyphenol content and antioxidant activity in methanol extract (80%) of common mistletoe (Viscum album L.) collected in April 2017 in Lašćina and Maksimir area in Zagreb (Croatia) from different plant host (linden - Tilia platyphyllos Scop., hawthorn - Crataegus monogyna Jacq. and maple - Acer campestre L.). To achieved that we performed reversed phase high- performance liquid chromatography (RP-HPLC) for individual detection of polyphenols and spectrophotometric Folin-Ciocalteu method for detection of total polyphenol content. Three different spectrophotometric methods for antioxidant activity determination ABTS (2, 2 – azinobis (3- ethylbenzothiazoline-6-sulfonic acid), DPPH (1, 1-diphenyl-2-picrylhydrazyl) and FRAP (Ferric Reducing/Antioxidant Power Assay) were also performed. One part of mistletoes leaves with their petioles were freeze-dried while other half was dried in shadow at room temperature (RT). Dried plant material was chopped into pieces with a diameter of approximately 5 mm end extract with 80% methanol in glass column (1 × 10 cm) for 24 hours. Percolate was discharged from the column at a rate of about 4 drops per minute and evaporated to dryness by rotavapor at 30 ° C. Methanol (80%) was added to give dry matter to solvent ratio of 1:1. With HPLC analysis of mistletoe samples was detected 8-10 phenolic compounds, of which 5-7 phenolic acids (p-coumaric, gallic, caffeic, chlorogenic, ferulic, synapyl and salicylic acid in mistletoe that grew on hawthorn, chlorogenic acid was not detected in mistletoe that grew on linden and maple and gallic acid was not detected in mistletoe that grew on linden) and 3 flavonoids (quercetin, kaempferol and isoramentin). Phenolic acids p-coumaric acid and sinapinic acid and antioxidant activity measured with ABTS and DPPH were statistically higher in freeze-dried than RT- dried plant material in almost all samples. The highest concentration of total and individual phenolics and generally highest antioxidant activity were detected in mistletoe samples that grew on hawthorn. Most mistletoe samples had strong antioxidant activity (> 70%) in relation to positive control Trolox. According to results, mistletoe that grew on hawthorn is the best source of polyphenols and antioxidants that can be use in phytotherapy for treatment number of different ailments. Also, freeze-drying has positive effect on antioxidant activity and preservation of p- coumaric acid and sinapinic acid in mistletoe samples.

ABTS, DPPH, FRAP, RP-HPLC, polifenoli

nije evidentirano

engleski

Phenolic content and antioxidant activity of mistletoe (Viscum album L.) from different plant host

Cilj ovog rada je ispitati sadržaj polifenola i antioksidacijsku aktivnost metanolnih ekstrakata (80%) bijele imele sakupljene u travnju 2017. na Lašćini i na području Maksimira u Zagrebu (Hrvatska) sa nekoliko različitih domadara (lipa - Tilia platyphyllos Scop. , glog - Crataegus monogyna Jacq. i javor - Acer campestre L). Tekućinskom kromatografijom visoke djelotvornosti s obrnutim fazama (RP-HPLC) detektirani su pojedinačni polifenoli, a količina ukupnih fenolnih spojeva određena je pomoću Folin- Ciocalteu metode. Tri različite metode korištene su za određivanje antioksidacijske aktivnosti ABTS (2, 2 –azinobis (3- ethylbenzothiazoline-6-sulfonic acid), DPPH (1, 1-diphenyl-2-picrylhydrazyl) i FRAP (Ferric Reducing/Antioxidant Power Assay). Dio listova imele sa peteljkama je liofiliziran dok je drugi dio sušen u sjeni i na sobnoj temperaturi. Osušeni biljni materijal usitnjen je u komadiće promjera približno 5 mm i ekstrahiran u 80 % etanolu u staklenoj koloni dimenzije 1×10 cm u trajanju od 24h. Perkolat je ispušten iz kolone brzinom od oko 4 kapi u minuti te uparen na rotavaporu na temperaturi od 30 °C . Metanol (80 %) je dodan kako bi se dobio omjer suhe biljne mase i otapala 1:1. HPLC analizom uzoraka listova imele detektirano je ukupno 8-10 fenolnih spojeva, od toga 5-7 fenolnih kiselina (p-kumarinska, galna, klorogenska, kavena, ferulična, sinapilna i salicilna kiselina u uzorcima imele koja je rasla na glogu, klorogenska kiselina nije detektirana u uzorcima imele koja je rasla na javoru i lipi, a galna kiselina nije detektirana u imeli koja je rasla na lipi) i 3 flavonoida (kvercetin, kempferol i isoramentin). Fenolne kiseline p- kumarinska i sinapilna te antioksidacijska aktivnost mjerena sa ABTS i DPPH metodom statistički značajno je bili viša u skoro svim liofiliziranim uzorcima u odnosu na uzorke imele sušene na sobnoj temperaturi. Najviša koncentracija ukupnih i individualnih fenola i generalno najviša antioksidacijska aktivnost detektirana je u uzorcima imele koja je rasla na glogu. Većina uzoraka imele imala je jaku antioksidacijsku aktivnost (>70%) u odnosu na pozitivnu kontrolu Trolox. Na temelju rezultata možemo zaključiti da uzorci imele koji su rasli na glogu predstavljaju najbolji izvor polifenola i antioksidanta koji se mogu primijeniti u fitoterapiji za liječenje različitih oboljenja. Također, liofilizacija ima pozitivan učinak na antioksidacijsku aktivnost (ABTS i DPPH) te na očuvanje koncentracije p-kumarinske i sinapilne kiseline u uzorcima imele./The aim of this study is investigated polyphenol content and antioxidant activity in methanol extract (80%) of common mistletoe (Viscum album L.) collected in April 2017 in Lašćina and Maksimir area in Zagreb (Croatia) from different plant host (linden - Tilia platyphyllos Scop., hawthorn - Crataegus monogyna Jacq. and maple - Acer campestre L.). To achieved that we performed reversed phase high- performance liquid chromatography (RP-HPLC) for individual detection of polyphenols and spectrophotometric Folin-Ciocalteu method for detection of total polyphenol content. Three different spectrophotometric methods for antioxidant activity determination ABTS (2, 2 – azinobis (3- ethylbenzothiazoline-6-sulfonic acid), DPPH (1, 1-diphenyl-2-picrylhydrazyl) and FRAP (Ferric Reducing/Antioxidant Power Assay) were also performed. One part of mistletoes leaves with their petioles were freeze-dried while other half was dried in shadow at room temperature (RT). Dried plant material was chopped into pieces with a diameter of approximately 5 mm end extract with 80% methanol in glass column (1 × 10 cm) for 24 hours. Percolate was discharged from the column at a rate of about 4 drops per minute and evaporated to dryness by rotavapor at 30 ° C. Methanol (80%) was added to give dry matter to solvent ratio of 1:1. With HPLC analysis of mistletoe samples was detected 8-10 phenolic compounds, of which 5-7 phenolic acids (p-coumaric, gallic, caffeic, chlorogenic, ferulic, synapyl and salicylic acid in mistletoe that grew on hawthorn, chlorogenic acid was not detected in mistletoe that grew on linden and maple and gallic acid was not detected in mistletoe that grew on linden) and 3 flavonoids (quercetin, kaempferol and isoramentin). Phenolic acids p-coumaric acid and sinapinic acid and antioxidant activity measured with ABTS and DPPH were statistically higher in freeze-dried than RT- dried plant material in almost all samples. The highest concentration of total and individual phenolics and generally highest antioxidant activity were detected in mistletoe samples that grew on hawthorn. Most mistletoe samples had strong antioxidant activity (> 70%) in relation to positive control Trolox. According to results, mistletoe that grew on hawthorn is the best source of polyphenols and antioxidants that can be use in phytotherapy for treatment number of different ailments. Also, freeze-drying has positive effect on antioxidant activity and preservation of p- coumaric acid and sinapinic acid in mistletoe samples.

ABTS, DPPH, FRAP, RP-HPLC, polyphenols

nije evidentirano