POLYPHENOL COLORATION OBJECTIVES 1. Polyphenols – classification and representatives; 2. Non-enzymatic polyphenol coloration; 3. Enzymatic polyphenol coloration;

POLYPHENOL COLORATION 1 Polyphenol coloration (non-enzymatic) – mostly due to complexation with metal cations, changes of the рН, disintegration or formation of some pigments during food processing. Polyphenol coloration (enzymatic) – due to action of polyphenol oxidases (PPO). ! Type of polyphenols Structure Type С6 Simple phenols С6 – С1 Phenolic acids and aldehydes С6 – С2 Derivatives of phenylacetic acid and acetophenone С6 – С3 Derivatives of cinnamic acid Coumarins and isocoumarins С6 – С3 – С6 Flavonoids С6–С1–С6; С6–С2–С6 Benzophenons, xanthons and stilbenes С6, С10, С14 Quinones Dimers and oligomers Lignans Polymer Lignin Tannins

POLYPHENOL COLORATION Phenolic acids and aldehydes 2 Simple phenols: resorcinol pyrogallol Phenolic acids and aldehydes р-hydroxy-benzoic protocatehuic vanillic salicylic gallic vanillin

POLYPHENOL COLORATION Derivatives of cinnamic acid 3 Derivatives of cinnamic acid 5-hydroxy-ferulic cinnamic caffeic sinapic р-coumaric ferulic Stilbenes – resveratrol

POLYPHENOL COLORATION 4 Lignans and lignin coniferyl alcohol р-coumaril alcohol Sinapyl alcohol Oligomeric and polymeric structures

POLYPHENOL COLORATION 5 Flavonoids General structure: γ-benzopyron linked to benzene ring flavonoids isoflavonoids neoflavonoids

POLYPHENOL COLORATION 6 Flavonoids Depending from the degree of saturation of the heterocycle (O-containing ring) the flavonoids could be: 1). Derivatives of phenylbenzo-γ-pyron (flavons, flavonols, flavanols and flavanonols) 2). Derivatives of phenylbenzo-γ-pyran (flavan-3-ols, anthocyanindins and flavan-3,4-diols) Flavons luteolin apigenin

POLYPHENOL COLORATION 7 Flavonoids Flavonols Flavonols beside the characteristic for the flavons keto-groups and C=C bond have OH group at 3rd place. Flavonols are much more distributed in nature than flavons. kaempferol quercetin myricetin

POLYPHENOL COLORATION 8 Flavonoids Flavanons The flavanons have keto-group but didn’t have C=C bond in the heterocycle naringenin Flavanonols taxifolin

POLYPHENOL COLORATION 9 Flavonoids Flavan-3-ols (cathehins) The name cathehin comes from catechu, which is the tannic juice or boiled extract of Mimosa catechu (Acacia catechu L.f). The cathehins could form esters with gallic acid – esterification of the OH group at 3’ position cathehin gallocathehin

POLYPHENOL COLORATION 10 Flavonoids Flavan-3,4-diols (leucoanthocyanidins) The leucoanthocyanidins are synthesized from flavanonols by reduction of the keto-group. They are found in the wood where they play a role in forming the condensed tannins. leucodelphinidin leucocyanidin

POLYPHENOL COLORATION ! POLYPHENOL COLORATION 11 Flavonoids Anthocyanidins In the nature the most often found are 6 anthocyanidins which after glycosylation give a great variety of anthocyanins (glycosides of anthocyanidins). They have pronounced P-vitamin activity, lower the fragility and permeability of the capillary. They have anti inflammatory, hypotonic and collagen-stabilizing action. The anthocyanins intake is beneficial for the capillary system and improvement of the vision. They play important role in the collagen crosslinking and inhibit its enzymatic degradation during inflammatory processes. Anthocyanins are applied successfully for prophylaxis and treatment of glaucoma and other ophthalmic diseases. In model systems extracts rich in anthocyanins show cardio-protective effect and inhibit the growth of cancer cells. Name R1 R2 Delphinidin OH Petunidin OCH3 H Cyanidin Pelargonidin Peonidin Malvidin

POLYPHENOL COLORATION 12 Complexation and color formation The flavonoids – found in almost all fruits and vegetables. Complexation with metals like Fe3+, Ca2+, Al3+, Sn2+, Cu2+ are the reason for coloration. Fe3+ - the complexes with tannins are used for tannins measurements in beers. The observed coloration of potatoes after boiling – complexation of Fe3+ with o-phenols. Sometimes coloration of processed cauliflower – complexes of Fe2+ kaempferol and quercetin. The canned cauliflower could give yellow colored complexes with tin. Formation of pink coloration (pinking) of pears and peaches after thermal treatments (and subsequent slow cooling) – due to formation of cyanidin complexes with copper, iron and zinc. More at CRC Handbook of Food Additives, Ed. Thomas E. Furia (2nd edition), CRC Press 1972 NY. (pages 281-283)

POLYPHENOL COLORATION 13 Complexation and color formation Coloration of canned asparagus shoots (dark-greenish to black) – complexes of rutin (quercetin-3-O-rutinoside) with Fe2+ (EDTA or presence of Sn2+ - eliminate the coloration problems). Generally use of sequestrants is the solution for complexation of polyphenols with metals and obtaining of unusual coloration or discoloration The formation of the color of ripe olives – dark-brown to black is due to formation of oxidative products of flavonoids (luteolin 7-glycoside). Other compounds responsible: peonidin-3-glucoside, cyanidin-3-rhamnoglucoside (acylated with caffeic acide), hydroxytyrosol, vanillic acid, caffeic acid, p-coumaric acid etc.

POLYPHENOL COLORATION 14 Changes of the color with change of the pH – example: anthocyanins Anthocyanins of red cabbage: left to right – increase of the рН

POLYPHENOL COLORATION 15 Anthocyanin structures Flavylium salt (red) Quinonoidal base (blue-dark green) Chalcone (colorless) Pseudobase or carbinol (colorless)

POLYPHENOL COLORATION 16 Stabilization of strawberry beverages with addition of polyphenols. Plamen Mollov, Kiril Mihalev, Vasil Shikov, Nikolina Yoncheva, Vasil Karagyozov, Colour stability improvement of strawberry beverage by fortification with polyphenolic copigments naturally occurring in rose petals, Innovative Food Science & Emerging Technologies, 8(3), 2007, 318–321.

POLYPHENOL COLORATION 17 Formation of complexes with phenolic compounds - stabilization Molecular complexes between anthocyanin and a polyhydroxyflavone sulfonate – Food chemistry, Fennema

POLYPHENOL COLORATION 18 Degradation of antocyanins Polymeric brown degradation products Glycoside bonds and presence of sugars – stabilizing effect on anthocyanins; but the products formed (cyclic aldehydes, carbonyls) when Maillard reaction takes place lead to condensation reaction with anthocyanins and formation of brown colored products

POLYPHENOL COLORATION 19 Interaction of phenolic compounds with 5-HMF (or HMF)

ENZYMATIC BROWNING 20

ENZYMATIC BROWNING 21 1. Significance - Formation of humus in the soil; - Biosynthesis of melanins in the organisms – pigmentation, defense from the UV-rays; - Formation of color in the tea leaves, tobacco, cocoa; processes of ripening of dates, figs, raisins etc.; - Protective mechanism of the fruit and vegetables; - Participate in the redox reactions – defence mechanism; Melanin in the melanocyte

ENZYMATIC BROWNING 22 2. Mechanism of action Polyphenol oxidase (РРО) Substrates of the reaction – monophenols, L-tyrosine, hydroxy-derivatives of cinnamic acid (p-coumaric, caffeic, ferulic, chlorogenic acid) Catechol oxidase – catalyzes oxidation of o-diphenols to quinones.

ENZYMATIC BROWNING 23 2. Mechanism of action

ENZYMATIC BROWNING 24 2. Mechanism of action – obtaining of melanins from L-tyrosine

ENZYMATIC BROWNING 25 3. Types of melanins 3.1. Alomelanins – plant melanins (catechol derived) catechol Sample structure of catechol melanins

ENZYMATIC BROWNING 26 3. Types of melanins 3.2. Eumelanins – animal type (tyrosine derived, indolic structure); dark-brown color. Tyrosine → DOPA → dopaquinone Dopachrome Leucodopachrome Sample structure of eumelanins

ENZYMATIC BROWNING 27 3. Types of melanins 3.3. Pheomelanins – animal type (tyrosine and cysteine derived, indolic structure); red-brown color. cysteine L-DOPA Sample structure of pheomelanins

! ENZYMATIC BROWNING 28 4. Processes of enzymatic browning desired - fermentation of tea leaves; - fermentation of cocoa beans; - obtaining of sake; - obtaining of soy sauce. undesirable - discoloration of wounded tissues – as a result of microbial attack, wounding during picking, transportation, storage; - treatment of the raw materials – peeling, trimming, milling, etc. - chilling and freezing processes; - drying processes; - processing of crustaceans. inhibition - Tropolone ( grape polyphenol oxidase inhibitor); - Potassium metabisulfite, K2S2O5; - Potassium dithionite or potassium hydrosulfite, K2S2O4;