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(15th): Toxicology and Phytochemistry
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(20th): Beverages and Fermentations  Thursday (22nd): No class  Tuesday (27rd): Additives, Radicals, Antioxidants  Thursday (29th): Exam #3. Last class day.
 Tuesday

Class Assignment #2 will be posted online TODAY.

Due date: On or before April 28th by 5 pm.

Follow the directions!

Assignment Topic:
  

Beverages Food Additives Legal Status

Food Toxicology

or narcotic effects. properties. and their impact on humans. stimulatory. and detection of toxic substances in food. .  Other plants were found to contain chemicals that have medicinal. people were aware that some plants are poisonous and should be avoided as food.  Early on. effects. hallucinatory.Food Toxicology  The study of the nature.

The Dose Makes the Poison Attributed to Pericles -a Greek statesman. .

“Toxic” to One….not to Another  Food Allergens (The “Big Eight”)  Cows milk  Shellfish (Crustacea)  Eggs  Fin fish  Peanuts  Soy  Tree nut  Wheat .

For example: Celiac Disease  Ingestion of wheat. bone pain. weakness. barley. rye  Proline-rich protein . chronic fatigue. weight loss.gliadins  Triggers immune damage to small intestine  Impairs absorption of nutrients  Diarrhea. bloating. anemia. muscle cramps .

.Scombroid Poisoning  Anaphylactic  Tuna. other pellagic fish species  Histamine  Also from spoilage bacteria in fish from putrecine and cadaverine  Everyone is susceptible  Some more sensitive (allergy?) than others. shock  Eating fish with high histamine levels mackerel.

Sensitivity or Allergy to:  Lactose  Sulfites  Strawberries (internal histamine release)  Fava beans/broad beans (enzyme deficiency)  Asparagus (sulfur compounds)  Red wine (low levels of histamine. SO2)  Fructose intolerance  Aspartame (phenylalanine)  Tartarazine (FD&C Yellow #5) .

000 species of fungi  > 300 mycotoxins isolated. 30 with food issues  Plant specific  Environmental  Temperature  Humidity  Moisture  Oxygen .Mycotoxins  Substances produced by fungi that are harmful to animals and humans  > 100.

. in wet and over-wintered grains: barley. wheat  Sclerotia or “ergots” (Hard-packed mycelium)  “Ergotism”  Convulsions and gastrointestinal symptoms  Ergotamine is an analogue of lysergic acid (LSD). a glycoalkaloid  Vasoconstrictor that may cause hallucinations  St. Anthony’s fire  Potential cause of Salem. 1692.Claviceps purpurea  Grows  rye. MA witch trials.

rice. transport. etc  Animal carcinogen at 5 ppb  Human liver carcinogen  Problem in food industry and grain handling  Harvesting. G2  In animal feed.Aspergillus flavus and A. pecans. wheat. paraciticus  Universal food  Corn. cow or human) . storage 4 main aflatoxins: B1. B1 and B2 can be converted to M1 and M2 and secreted in milk (ie. contaminant peanuts. B2. G1. walnuts.

Toxins formed during Food Processing  We previously covered the Maillard reaction    Polycyclic aromatic hydrocarbons (benzo(a)pyrine) Carcinogenic agents in almost every model tested But excellent flavor compounds.yet remember Pericles?  Nitrite used in curing meat and fish     Antimicrobial agent Reacts with myoglobin and hemoglobin to form red nitrosyl compounds Nitrite may also react with amines to form nitrosoamines. .. Carcinogenic. mutagenic..

Higher in high carbohydrate foods. cosmetics. etc)  Broad range of foods with significant levels of acrylamide. thickening agent for pesticides. but not boiled. soil conditioner.  Used as:  Cement binder. laboratory gels.  Acrylamide is a neurotoxin and carcinogen. plastic manufacture. waste water treatment agent. . Fried and baked.    Foods prepared at high temperatures.Toxins formed during Food Processing  Acrylamide in foods  In 2000-2002 Swedish researchers identified acrylamide in foods and residues from human samples.

Acrylamide in Potatoes  Acrylamide derived from asparagine in the presence of sugar.  Carbonyl carbon in glucose facilitates the reaction  Asparagine + Sugar + Heat = Acrylamide .

Natural Food “Toxins” in Plants .

ect  Includes the production of plant primary and secondary compounds. carbon dioxide.Metabolism and Plant Primary Compounds  Plant metabolism:  Used  to describe the chemical reactions and interactions that take place in a biological system. and water. .  Plant Primary Compounds  Those compounds which are formed as a part of the normal anabolic and catabolic processes. Includes reactions that cause a plant to shift its leaves towards a light source  Cause chloroplasts to produce photosynthetic sugars from sunlight.

 At least one major benefit a plant gains from producing secondary compounds was later discovered: protection from certain types of herbivory.Plant Secondary Compounds  Secondary Compounds  Those compounds that are not primary compounds.  The bitter taste of tea is due to the secondary compounds found in the tea leaves (caffeine and tannins).  Some regarded these substances as waste products of a cell.  The most obvious way to discourage this was to synthesize a substance to make the plant taste bad. .  A compound that does not seem to directly function in the processes of growth and development.

or difficulties in labor. etc) . “weed”. nutmeg. it mimics the hormone progesterone and may cause infertility.  Not so coincidentally. coffee. Found in alfalfa.  Isoflavonoids affect reproduction in mammals. and some clover species. this same type of physiological effect is what people refer to as the "high" after consuming certain plants (mushrooms. reduced lactation.Other Mean Tricks  Secondary Compounds  Another way to hinder herbivory was to synthesize compounds that would cause temporary or permanent physiological changes in the herbivore (Toxins)  Caffeine and morphine are good examples. soy.  These types of secondary compounds make an animal "think twice" about eating off the same plant again.

Do plants experience stress the way humans do? Hypericum perforatun Lycopersicum esculentun .

transport)  Microbial contamination (polysaccharides) . hail)  Ethylene (ppb levels)  Post-harvest storage  UV light  Physical wounds (harvest. wind. flood.Causes of Plant Stress (Elicitors)  Insects (US est. at $100M annum)  Weather (drought.

000 times more “natural” chemicals than man-made chemicals  ~150 true phytoalexins known .Results of Plant Stress Phytoalexins (Phyto = “plant” and alexin = “to ward off”)  Low MW organic metabolites produced by plants in response to stress  “Self-made antibiotics”  We consume ~10.

astringent and sour flavors .What? Toxic Chemicals in My Food!! Phytochemical Paradox  Dangerous pesticide or antioxidant wonder?? For the plant:  UV light protection (Nature’s sun-screen)  Oxidative protection  Anti-microbial (Anti-Herbivores?) For the consumer (us):  ROS quencher  Bitter.

DEFENSE MECHANISMS Pre-existing defenses Induced/Biochemical defenses Hypersensitive responses .

PRODUCTION OF PHYTOALEXINS Production of phytoalexins may be stimulated by certain compounds called elicitors. High molecular weight substances found in the cell wall such as glucans. or other polysaccharides. Gases such as ethylene (C2H4) . glycoprotein.

The suppressor also can be a glucan. or a toxin produced by the pathogen .PRODUCTION OF PHYTOALEXINS In susceptible plants. by the action of suppressors produced by the pathogen. a glycoprotein. a pathogen may prevent the formation of phytoalexins.

Chemical and Sensory Properties of Food “Toxins”  Aromatic  Sour  Bitter  Astringent  Sulfurous  Medicinal  Smoky  Animal-like  Metal chelator  Antioxidant (-OH groups)   Free radical terminators ROO + AH = ROH + A . . . O O O .  Resonance stability O OH . .

Phytoalexic Toxins In Our Food .

Potato Stress Metabolites  Cinnamates. solanine. chaconine Solanine (Glycoalkaloid)  Sunburned spuds or growth shoots (periderm)  10-50 ppm is normal. sesquiterpenoids. increases 7-fold during stress  Natural pesticide (cholinesterase inhibitor)  Acetylcholine is a neurotransmitter  Extremely bitter. quinic acid. not soluble in water  Heat stable . scopolin.

Tomato Stress Metabolites  Cinnamates. tomatine Tomatine (Alkaloid)  High in immature fruit  Ripe tomato contains ~30-40 ppm  Natural pesticide  Heat labile . falcarindiol. rishitin.

falcarinol. isocoumarin Isocoumarin (neutral phenolic)  Anti-microbial (~10 ppm)  Extremely bitter.Carrot Stress Metabolites  Cinnamates. not soluble in water  Heat stable  Ethylene sensitive synthesis . falcarindiol.

5.“Natural” Carcinogens in Coffee          Acetaldehyde Benzaldehyde Benzene Benzofuran Benzo[a]pyrene Caffeic acid Catechol 1.6 Dibenzanthracene Ethanol  Ethylbenzene  Formaldehyde  Furan  Furfural  Hydrogen peroxide  Hydroquinone  Limonine  Styrene  Toluene  Xylene .2.

Medicarpin (Isoflavonoid) Soybean roots.Capsidiol Sweet potato.Phaseollin .Glyceollin Peas.Psoralens (furanocoumarins) Grapes.Other Commodities Peppers.Ipomeamarone Celery.Resveratrol. parsley.Pisatin Bean pods. parsnips. stilbene Alfalfa.

GI tract  Enzyme system to solubilize chemicals  Substrate (xenobiotics) must be somewhat lipophilic to reach active sites for metabolism  Plants did not want to become food for people or animals…. nasal mucosa.but we fought back !! . lung.Plant vs Man: “Xenobiotic” Metabolism Cytochrome P-450 system  In body: liver. skin.

“Antioxidant” Enzyme Systems  Active  SOD Oxygen Detoxification Enzymes (superoxide dismutase)  GSH (glutathione)  POD (peroxidase)  CAT (catalase) .

Add/Remove H. . ie. -Tend to increase size and polarity for excretion.Cytochrome P-450 system Induction of Phase I and Phase II Enzymes Phase I Oxidation/Reduction – Add/Remove O. alcohol dehydrogenase Hydrolysis. – Add water Phase II Conjugation reactions. -Enzyme catalyzed reactions.


Current trend: Increased consumer interest in nutraceuticals…..which are phytochemicals

Dietary compounds, foods, or supplements used to promote health life or prevent some disease

Components in a plant based diet other than traditional nutrients that can potentially reduce the risk of degenerative diseases

Vit K and E





Phyto= plant…chemicals  Organosulfides  Isothiocynates  Indoles  Carotenoids  Saponins  Tocopherols  Amino acids/Proteins  Lipids  Carbohydrates  Polyphenols    Flavonoids Tannins Isoflavones  Vitamins/Minerals  Coumarins  Dietary Fiber  Enzymes .

nitrosamines)  Alter estrogen metabolism  Decrease cell proliferation  Maintenance of normal DNA repair .Mechanisms of Action  Many antioxidant effects  Increased activity of enzymes that detoxify carcinogens  Effect on cell differentiation  Block formation of carcinogens (ie.

How Did They Get There ?? Selective Biosynthesis .

How are Phytoalexins Formed? The “-noids”  Shikimic acid pathway (phenylpropanoids)    Hydroxycinnamic acids Coumarins Hydroxybenzoic acids OH OOCCH HOOC HO OH OH C H OH  Mevalonic acid pathway (Isoprenoids)   Carotenoids Terpenoids OH OH HO O OH OH O  Combination of Pathways (Shikimic-Polymalonic)  Flavonoids and anthocyanins .

Shikimic Acid PathwayPhenolics  PEP from glycolysis + erythrose 4-P from PP-pathway forms skikimic acid which are the precursor to phenylalanine and tyrosine.  PAL and TAL (ammonia lyases)  Cinnamic  Coumarin HO COOH HO COOH acid HO MeO  p-Coumaric  Caffeic acid HO COOH acid  Ferulic acid  Sinapic acid MeO HO COOH MeO .


Leucocyanidin R1 = R2 = OH. Cyanidin-3-glycoside R1 = R2 = OH. Pelargonidin-3-glycoside R1 = H. Hesperitin O HOOC SCoA acetyl CoA carboxylase Me O SCoA S-adenosyl-homocysteine S-adenosyl-methionine OH R2 OH HO O + R2 UDP R1 Gly UDP-Dglucose HO O + 2H2O O2 & alphaOH CO2 succinate ketoglutarate R1 R1 = R2 = H. Apiferol R = OH. Diosmetin S-adenosyl -methionine S-adenosyl -homocysteine isoflavone methyl transferase S-adenosyl -homocysteine OH OH Flavan-4-ol HO O OH H H2O HO O NADPH NADP+ O2 H2O HO isoflavone hydrolase OH O OH O O OH O methoxyisoflavone dehydratase OH O O 2. Dihydroquercetin R1 = R2 = OH. Myricetin Flavone R1 = OH. R2 =OH. R2 = H. Fortuneletin R1 = R2 = OH. Luteoferol OH 2-hydroxyisoflavanone naringenin S-adenosyl -methionine OH O OH Prunetin Flavonol R1 = R2 = H. R2 = H.7-dihydroxy-4'methoxyisoflavanone HO o-methyl transferase S-adenosylhomocysteine COS-CoA O O protocatechuic acid hydroxylase OH p-hydroxybenzoic OH acid hydroxylase OH protocatechuic acid p-hydroxybenzoic acid HO p-hydroxybenzaldehyde S-adenosylmethionine HO O OH OH O OH -3CO2 R1 = R2 = H. R2 =H. R2 =OH. Delphinidin R1 = OH.4-diol (leucoanthocyanidin) HO R1 OH flavanone 3-hydrolase O R1 chalcone isomerase HO OH NADPH O2 NADP+ HO H2O O 2-hydroxyisolflavanone dehydratase flavonoid glucosyltransferase OH OH Anthocyanidin OH Anthocyanidin3-O-glycosides anthocyanidin synthase dihydroflavonol 4-reductase OH O isoflavone synthase OH O OH O OH O O Liquiritigenin OH O OH Dihydroflavonol Flavanone Chalcone Isoliquiritigenin 2-hydroxyisoflavanone Flavonoids HO O OH HO O leucoanthocyanidin reductase R2 R4 OH R3 R2 OH R1 OH OH Procyanidins leucoanthocyanidin reductase OH HO O flavonol synthase R2 OH flavone synthase I & II R2 OH HO O R1 OH O Naringenin OH NADPH O2 NADP+ H2O HO isoflavone synthase O OH H2O isoflavone dehydratase HO O flavanone 4-reductase HO O S-adenosyl S-adenosyl -methionine -homocysteine isoflaveone methyl transferase O O HO O OH OH Flav-3-nol R1 = R2 = H. Delphinidin-3-glycoside R1 = R2 = H. Dihydrokaempferol R1 = H. R2 = H. Catechin R1 = R2 = OH. Cyanidin R1 = R2 = OH. R2 =OH. R2 =OH. Leucodelphinidin chalcone synthase OH o-methyl transferase H2O Daidzein Malonyl CoA OH OH chalcone HO reductase chalcone isomerase OH isoflavone synthase HO O R2 OH NADP+ NADPH HO R1 O R2 OH R2 HO O OH OH OH Flavan-3. Gallocatechin R1 OH OH O OH O genistein OH R1 OH O OH O HO O OH R R = H. R2 = OH. Pelargonidin R1 = H.5. Apiginin R1 = OCH3. Eriodictyol R1 = OCH3. Luteolin R1 = OCH3. R2 = OH. Kaempferol R1 = H. R2 =OH.5 cafeoylquinic acid) OH OH quinic acid HO H Arogenic acid glutamine 2-oxoglutarate amidotransferase (GOGAT) glutamine glutamic acid OH OH OH OH OH Chlorogenic acid (5 cafeoylquinic acid) OH Isochlorogenic acid (4. R2 =OH. Dihydromyricetin R1 = R2 = H. Naringenin R1 = OCH3. Afzelechin R1 = H.The Shikimic Acid Pathway Leading to Phenylpropanoid and Flavonoid Synthesis OH HO HO OH D-glucose O OH O3PO H HO O COO+ O3PO DAHP synthetase HO O O3POH2C COOH HO dehydroquinate synthase COOH dehydroquinate dehydratase COOH NADPH NADP+ COOH COOH ATP ADP O3PO OH 3-phosphoshikimic acid OH COOH H H H COOH O3PO PEP EPSP synthase COOH COOH OH erythrose 4-phosphate OH O OH O OH phosphoenolpyruvate shikimate dehydrogenase HO OH shikimate kinase OH deoxy-arabinoheptulosonate-7-phosphate OH 3-dehydroquinic acid OH 3-dehydroshikimic acid OH shikimic acid O3PO OH O COOH OPO3 O3PO OH O COOH chorismate synthetase O COOH quinate dehydrogenase HO COOH HO COOH 5-enolpyruvyl shikimic acid 3-phosphate OH chorismic acid COOH OH HO chorismate mutase COOH HO OH quinic acid OH 3OPO OH O prephenate dehydratase COO- OH 3-phosphoquinic acid HOOC Phenylpyruvic acid COOH HC NH2 HOOC arogenate dehydratase CH2 2-oxoglutaric acid glutamic acid HO O O OH COOH HO O O COOH HO COOH HO HO O OH COOH COOH NH4+ H NH3+ prephenic acid OH O O O OH O HO O OH O Cinnamic acid L-phenylalanine ammonia lyase (PAL) O HO COOH Phenylalanine (phe) HO OH OH OH OH Isochlorogenic acid (3. Isosakuranetin R1 = R2 = OH. Quercetin R1 = R2 = OH. Leucopelargonidin R1 = H.5 cafeoylquinic acid) Chlorogenic acid (4 cafeoylquinic acid) NADP+ NH4+ from PAL cinnamate4-hydroxylase NADPH arogenate dehydrogenase COOH COOH Phenylpropanoi ds MeO HO OMe Sinnapic acid Caffeic acid 3-O-methyltransferase HO HO OMe COOH MeO Ferululate-5 -hydroxylase COOH caffeate methyl3-O-transferase HO HO Caffeic acid COOH p-comuarate 3-mono-oxygenase COOH tyrosine ammonia -lyase (TAL) NH2 HO O HO Ferulic acid S-adenosyl-methionine HO p-coumaric acid 5-hydroxyferulic acid NH4+ OH tyrosine OH O OMe Formononetin H2O hydroxycinnamate:CoA ligase (4-coumaroyl:CoA ligase) COOH H2O NADPH HO OH gallic acid OH O2 NADP+ COOH H2O NADPH O2 NADP+ COOH NADPH OH NADP+ CHO COOH HO p-coumaryl-CoA O 2.7-trihydroxy-4'-methoxyisoflavanone Biochinan A O Pratensein .

Phenolics  Plants produce a variety of compounds that contain one or more phenol groups called phenolics  Thousands of phenolics occur in plants .

Phenolics  Large group of diverse compounds  Many serve as defense compounds against herbivores and pathogens  Some attract pollinators  Some absorb UV light  Some reduce growth of competitors .

Background  Flavonoids are non-nutrients  1936-Szent-Gyorgyi.  1950’s disproved the theory  Late seventies-mutagenecity of quercetin  Recent research-anticarcinogenic . called flavonoids Vitamin P.

Structure of flavonoids R1 31 21 B HO 8 O 2 A 6 C 4 3 X 61 41 OH 7 51 R2 5 HO O .

FLAVONOLS Kaempferol Quercetin Myrcetin .

Kaempferol OH HO O OH HO O .

Quercetin OH HO O OH OH HO O .

Myricetin OH OH HO O OH OH HO O .

Two Specific Polyphenolics Ellagic acid Isoflavonoids .

2003 .al. Mullen et.Ellagic acid Derivatives Ref.

Ellagic acid Conversion .

17 b. 2. 3.estradiol Presence of a phenolic (benzene) ring: Prerequisite for binding to the estrogen receptors.groups separated by a similar distance Common compounds: genestein/genestin and diadzein/diadzin . Pair of OH.Isoflavones share several features in common with estradiol OH OH O OH CH3 HO O HO Isoflavone 1.

and “Conjugated” Forms R1 R2 HO O OH O R1 O OH Aglycone OH H2C H H HO R2 O H O H O OH H OH H OH R1 R2 O Glycoside H R3 O H2C O H O H O H HO H OH H Acetyl  COCH3 Malonyl  COCH2COOH OH .Isoflavones: Free.

Phytochemicals Carotenoids .

 Acetate + Acetyl CoA = Acetoacetyl CoA  Acetoacetyl CoA + Acetyl CoA = Mevalonic acid     Phytoene Lycopene ß-carotene Lutein .Mevalonic Acid Pathway-Carotenoids  Pyruvate from glycolysis is decarboxylated to form acetate which combines with coenzyme A to form acetyl CoA.

The Mevalonic Acid Pathway for Carotenoid Synthesis Glyceraldehyde 3-phosphate thiamine pyrophosphate COO H Pyruvate NADPH + H+ CO2 H OH HO TPP 2OPO3 NADP+ HO HO OPO 3 OH 2-C-methyl-D-erythriol 4-phosphate 2- O 2OPO 3 OPO 3PO 3 metabolites unknown Isopentyl diphosphate (IPP) IPP Isomerase OPO3PO3 Dimethallyl pyrophosphate IPP Isomerase O 1-deoxy-D-xylulose 5-phosphate synthase OH 1-deoxy-D-xylulose 5-phosphate IPP Isomerase IPP Isomerase OPO3PO3 Geranylgeranyl diphosphate (GGPP) OPO3PO3 Geranyl diphosphate (GPP) OPO3PO3 Farnesyl diphosphate (FPP) phytoene synthase Prephytoene pyrophosphate phytoene synthase Phytoene phytoene desaturase Phytofluene carotene isomerase phytoene desaturase Zetacarotene zetacarotene desaturase Neurosporene zetacarotene desaturase Lycopene lycopene epsilon cyclase lycopene beta cyclase Delta-carotene Gamma-carotene lycopene beta cyclase lycopene beta cyclase Alpha-carotene Beta-carotene beta-carotene hydrolase beta-carotene hydrolasee OH Zeinoxanthin alpha-carotene hydrolase OH HO Beta-cryptoxanthin beta-carotene hydrolase OH HO Lutein HO Zeaxanthin .

Nomenclature and Structure Beta-Carotene HO Beta-Cryptoxanthin OH HO Lutein .

Metabolism of Dietary Carotenoids .

Breeding Fruits and Vegetables for Functionality .

Phytochemicals for Improved Human Health  Crops containing or that can be developed with:  Anthocyanins  Carotenes  Phenolic acids  Quercetin  Vitamins C. and E . D.

Blueberry Antioxidants 90 80 70 60 ORAC Anthocyanins Total Phenolics Chlorogenic acid mg/10g 50 40 30 20 10 0 Bluecrop Tifblue Duke Reveille Summit .

Blackberry Antioxidants 160 140 120 mg/10g 100 80 60 40 20 0 Apache ORAC Anthocyanins Total Phenolics Ellagic acid Chickasaw A-1960 Shawnee Choctaw .

(10) Tomato (10) 100 120 140 .62 Total Phenolics (mg/kg Chlorogenic acid) 6000 4500 3000 1500 0 0 20 40 60 80 ORAC (µM Trolox equivalents) Blackberry (13) Blueberry (9) Grape (10) Peach-Nectar.Total Soluble Phenolics and ORAC 7500 R2 = 0.

51 (mg/kg Malvidin 3-Glu) 1600 1200 800 400 0 0 20 40 60 80 100 120 140 Anthocyanin Blackberry (13) Blueberry (9) Grape (4) Nectarine (1) ORAC (µM Trolox equivilents) .Anthocyanins and ORAC 2000 R2 = 0.

Structure-Based Antioxidant Activity of Phytochemicals .

HO 0.22 HO COOH COOH 1.08 HO HO MeO HO COOH p-OH-benzoic p-coumaric HO 2.19 Protocatechuic Caffeic 1.43 MeO HO MeO COOH 1.26 HO MeO COOH COOH Vanillic Ferulic HO 1.90 MeO HO COOH COOH MeO .

Structurally Similar Compounds OH OH HO O OH OH O Quercetin AOX = 4.4 OH .7 HO + O OH OH OH HO O OH OH OH Catechin AOX = 2.4 Cyanidin OH AOX = 4.

Importance of the 3-OH group OH OH HO O OH OH O HO OH OH HO O O O OH O O O O Quercetin-3-glucoside AOX ~ 2.7 OH O Luteolin AOX = 2.1 .5 O OH OH O Quercetin AOX = 4.

Importance of the 2-3 db OH OH HO O OH OH O OH O HO O OH OH OH Quercetin AOX = 4.9 .7 Taxifolin AOX = 1.

7 HO O OH OH O OH O .9 OH OH OH O OH HO O Quercetin AOX = 4.1 OH Kaempferol AOX = 1.4 Luteolin AOX = 2.OH OH HO O Rutinoside OH O Summation HO O OH OH O OH OH HO O OH OH Rutin AOX = 2.3 Taxifolin AOX = 1.