Berry press residues as a valuable source of polyphenolics: extraction optimisation and analysis

Transcription

1 Berry press residues as a valuable source of polyphenolics: extraction optimisation and analysis Maris Klavins, Agnese Kukela, Linards Klavins, Jorens Kviesis

2 G.Arcimboldo Louvre Application possibilities of fruits and berries

3 Berry press residues as a source of valuable compounds Berry press residues (pomace) often is a waste material Berry press residues can serve as a source of many biologically active substances with diverse fields of applications One of most prospective groups of substances - polyphenolics

4 Study object Vaccinium berries Vaccinium myrtillus L Vaccinium uliginosum L Vaccinium corymbosum L Vaccinium vitis-idaea L Vaccinium oxycoccos L Vaccinium macrocarpon Blueberry Bog bilberry Highbush blueberry Lingonberry Bog cranberry Large cranberry

5 Comparison of different extraction methods Method Dry residue, g 100/g berry powder Total carbohydrates g 100/g berry powder Anthocyanins, g 100/g berry powder Total polyphenols, g 100/g berry powder Microwave ± ± ± ± 0.04 Soxhlet ± ± ± ± W ultrasound ± ± ± ± W ultrasound ± ± ± ± h shaking ± ± ± ± 0.06 Supercritical CO ± ± 0.01 ND 0.05 ± 0.01

6 Comparison of solvent systems Extrahent Dry residue, g/100 g Carbohydrate s, g/100 g Anthocyanines, g/100 g Polypheno lics, g/100 g CH 3 CN 49.5%, TFA 0.5%, H 2 O 50% ± ± ± ± 0.12 Acetone 50% ± ± ± ± 0.08 Acetone 75% ± ± ± ± 0.08 Methanole 60%, acetone 30%, H 2 O 10% ± ± ± ± 0.07 Methanole, HCl 1% ± ± ± ± 0.14 H 2 O, HCl 1% ± ± ± ± 0.03 Ethanole 70%, HCl 1% ± ± ± ± 0.09

7 Total anthocyanines, g/100 g Total polyphenolics, g/100 g Acidification impact on total polyphenol and anthocyanin yields 0,300 0,250 0,200 0,150 0,100 0,050 0,000 3,0 2,5 2,0 1,5 1,0 0,5 0,0 Total anthocyanins Total polyphenols

8 Total anthocyanins, g 100 g-1 berry powder 0,200 Kinetics of anthocyanine extraction from cranberry press residues 0,150 0,100 0, Ultrasound treatment, minutes

9 Total polyphenols, g 100 g-1 berry powder Kinetics of polyphenol extraction from cranberry press residues 3,0 2,8 2,5 2,3 2,0 1,8 1,5 1,3 1, Ultrasound treatment, minutes

10 Total anthocyanines, g/100 Total polyphenols, g/100 g Effect of the solvent/solid ratio on the extraction yield 0,200 3,0 0,150 2,5 2,0 0,100 1,5 0,050 1,0 0,5 0,000 1:30 EtOH 1:60 EtOH 1:90 EtOH 1:120 EtOH 1:30 MeOH 1:60 MeOH 1:90 MeOH 1:120 MeOH 0,0 Total anthocyanins Total polyphenols

11 Optimization of extraction conditions Optimization of extraction of phenolic compounds and anthocyanins from American cranberry press residues in aqueous ethanol and methanol with TFA and formic acid was carried out using Response Surface Methodology. A two factor and three level central composite design consisting of eleven experimental runs was employed. Coded levels Independent variable Symbol Ethanol/Methanol, v/v % X 1 96% / 100% Formic acid/tfa, v/v % X 2 10% / 1% 5% / 0.5% 1% / 0.1%

12 Two factor and three level Central Composite Design plan Factor 1 (X 1 ) Factor 1 (X 2 ) Ethanol, v/v% TFA, v/v% (code) (code) Standard order Run order Level code (+1) 0.1 (+1) (+1) 0.5 (0) (+1) 1.0 (-1) (0) 0.1 (+1) (0) 0.5 (0) (0) 0.5 (0) (0) 0.5 (0) (0) 1.0 (-1) (-1) 0.1 (+1) (-1) 0.5 (0) (-1) 1.0 (-1) -1-1

13 Response surface optimization of polyphenol extraction

14 Response surface optimization of anthocyanine extraction

15 Anthocyanins, mg/100g berries Anthocyanins, mg/100g berries Results of optimization: whole berries vs. press residues Lingonberry Blueberry MeOH EtOH MeOH EtOH Whole bery Optimal conditions Control conditions Press residue MeOH EtOH MeOH EtOH Whole bery Optimal conditions Press residue Control conditions

16 Purification of extracts to obtain purified polyphenolics 1 Berry pomace Extraction at optimal conditions 2 3 Reextraction to remove lipids Column chromatography (XAD 7) to remove carbohydrates Evaporation, lyophilisation

17 Yields and radical scavenging activity of obtained extracts Yields, g/100g FRAP ABTS DPPH Sample pomace mm Trolox eq/1g Large cranberries Highbush blueberries Bog cranberries Blueberries Lingonberries Control: ascorbic acid Studied anthocyanines

18 Chromatograms of berry anthocyanines Blueberries Bilberries Cranberries Lingonberries

19 Concentrations of anthocyanines in extracts (UPLC-PDA) N peak Compound Blueberry Billberry Cranberry Lingonberry American cranberry 1 delphinidin-3-o-galactoside ± ± delphinidin-3-o-glucoside ± ± cyanidin-3-o-galactoside ± ± ± ± ± delphinidin-3-o-arabinoside ± ± cyanidin-3-o-glucoside 0.71 ± ± ± ± ± petunidin-3-o-galactoside 7 cyanidin-3-o-arabinoside ± ± ± ± ± petunidin-3-o-glucoside 8.95 ± ± peonidin-3-o-galactoside 2.73 ± ± ± ± ± petunidin-3-o-arabinoside 8.55 ± ± peonidin-3-o-glucoside 3.28 ± ± ± ± ± malvidin-3-o-galactoside 13 peonidin-3-o-arabinoside 9.93 ± ± ± ± ± malvinidin-3-o-glucoside ± ± malvidin-3-o-arabinoside 6.39 ± ± 1.14

20 LC-HRMS (Orbitrap) results μg/g Compound Lingonberry Cranberry American Highbush Blueberry Crowberry cranberry blueberry 4-hydroxybenzoic acid Protocatechuic acid p-coumaric acid Gallic acid Caffeic acid Ferulic acid Syringic acid Resveratrol Pelargonidin Naringenin Kaempferol Cyanidin (+)-Catechin (-)-Epicatechin Ellagic acid Quercetin Delphinidin Taxifolin Myricetin Malvidin Chlorogenic acid Quercetin-3-glucoside Procyanidin A Procyanidin B Rutin Total amount of phenolics, μg/g

21 Thank you for attention! This work has been supported by the European Regional Development Fund within the project No /16/A/047 Genus Vaccinium berry processing using "green" technologies and innovative, pharmacologically characterized biopharmaceutical products.