Flavonoid D Class 2024 PDF

 More than 2000 plant-derived flavonoids have been identified.  Flavonoid compounds are the largest group of phenols found in nature.  The term flavonoid is derived from the word flavone, which is one of the most common types of flavonoids found besides flavonols and anth...

 More than 2000 plant-derived flavonoids have been identified.  Flavonoid compounds are the largest group of phenols found in nature.  The term flavonoid is derived from the word flavone, which is one of the most common types of flavonoids found besides flavonols and anthocyanidins.  Flavones have a 2 phenyl chroman skeleton  Based on the degree of oxidation, flavanes are the lowest and are used as the parent of the flavone name. 1 B O O O 2 A C 3 OH 4 katecin 2-fenilkroman flavan (flavan-3-ol) O OH O OH O leukoantosianidin calkon dihidrocalkon (flavan-3,4-diol)  These compounds play an important role in determining color, taste, odor  Flavonoids are often present in epidermal cells.  Most flavonoids are accumulated in the cell vacuolesel  Light, especially blue wavelengths, can increase flavonoid formation  Flavonoids also increase plant resistance to UV radiation  O O O O OH OH O O O O flavanon flavanonol flavon flavonol + + O O O C H OH O garam flavilium antosianidin auron  Anthocyanin - berries  Flavanone - citrus  Flavanol - red wine teas chocolate fruit  Flavonol - fruit vegetables  Hydroxycinnamates - most fruit & some vegetables  Absorption and Biotransformation of Dietary Flavonoids In Vivo Dietary SKIN AND Flavonoid BRAIN Oligomers Stomach cleaved cells Oligomeric Monomeric Flavonoids units Phase I and II O-methylated jejunum metabolism Further metabolism Sulphates Small Intestine Portal vein glucuronides ileum Liver glucuronides Kidney Colon Flavonoid Phenolic acids Renal excretion Gut microflora of glucuronides Urine Catechin enter to the bodies orally and absorbed by intestine, accumulated in liver Catechin phenolic groups conjugation Sulfuric acid Glucoronic acid 3-O- catechin-sulphat Epicatechin- 7-O- glucoronide Donor ion H+ to Pb radical, than lipid peroxidation is stopped GST activity ( GST chelating Pb) Pb excreted by urine and faeces FlavOH + ferryl radical  FlavO. Phenoxyl radical. FlavO + GSH.  GS Thiyl radical GS. + O2  Reactive oxygen species  GSSG Galati et al. 2002 FLOH + R* FLO* + RH FLO* + R* FLO – R FLO* + FLO* FLO-OFL Information: FLOH : flavonoid FLO*: phenolic flavonoid radicals R* : free radical Catechin activated GPx +R * Free radical lipid Peroxide radical H2O2 + 2 LH + 2 GSH GPx 2 LH + GSSG + H2O 2 H2O2 +LOOH + GSH Gpx LOH + GSSG + 2 H2O  Less bioavailable than ascorbate and tocopherols  MODIFIED by metabolism on absorption  Less extensively absorbed and circulating levels in vivo much lower Flavan-3-ol: METHYL + SULPHATE +GLUCURONIDE Wine catechins 100 nM Procyanidin: 4 uM; 0.26 uM; EPICATECHIN SULPHATE + Chocolate/cocoa 0.7 uM GLUCURONIDE Flavanone – NARINGENIN /HESPERETIN grapefruit/orange < 4 uM GLUCURONIDE Anthocyanin – 100 nM; 147 nM ANTHOCYANIN GLYCOSIDES berry juices Donovan et al., Keen et al., Baba et al., Ameer et al, Miyazawa et al.  Derived from Greek: anthos (flower) and kyanos (dark blue) colored pigments which are commonly found in red, purple and blue flowers.  This pigment is also found in various parts of other plants such as certain fruits, stems, leaves and even roots  Anthocyanin is a water-soluble pigment that is naturally present in various types of plants. As the name implies, this pigment gives color to the flowers, fruits and leaves of green plants, and has been widely used as a natural dye in various food products and various other applications  The color of the anthocyanin occurs because of the long conjugated double bond arrangement, so as to absorb light in the range of visible light. This conjugated double bonding system is also able to make Anthocyanin as an antioxidant with a radical arrest mechanism  BeberaMpa senyawa antosianin yang most found are :  pelargonidin, peonidin, sianidin, malvidin, petunidin, dan delfinidin.  Anthocyanin and other flavonoids attract the attention of many geneticists because by knowing the types of flavonoids they contain, it is possible to link various morphological differences between species that are closely related in one genus  Most plants have the largest anthocyanin content in the fruit Some other plants, such as tea, cocoa, cereals, beans, red cabbage and petunias also contain anthocynins in other parts of the body than fruit.  Grapes are the most widely used as a source of anthocyanins because the pigment content is quite high in the skin of grapes. So that the wine skin of the rest of the wine-making industry is often collected again to extract its anthocyanin content with acidic solvents.  pH Cation Oxygen o Sulphur dioxide (SO2) Protein Enzyme  pH The color caused by anthocyanins depends on the level of acidity (pH) of the surrounding environment so that this pigment can be used as a pH indicator.  The colors are red (pH 1), reddish blue (pH 4), purple (pH 6), blue (pH 8), green (pH 12), and yellow (pH 13).  To get the desired color, anthocyanins must be stored using a suitable pH buffer solution.  Cation Some cations, especially divalent and trivalent cations can cause anthocyanin discoloration to blue until pigment deposition occurs. In addition, the surface of copper, mild steel, and iron should also be avoided. Oxygen When dissolved in a mixed solution, anthocyanins will oxidize slowly.  Sulfur dioxide (SO2) If sulfur dioxide reacts with anthocyanin, a colorless product will form.  The color change reaction is reversible so that only by heating SO2 will the color return to its original state.  Protein If the source of anthocyanin reacts with protein, steam or sediment will form.  This event is more influenced by non-phenolic pigments that react with proteins such as gelatin.  Enzyme  The use of several enzymes in the processing of foods containing anthocyanins can cause anthocyanin candles in them to disappear or decrease.  This is partly due to the glucosidase enzyme which is present in enzyme preparation.  Various types of anthocyanin pigments extracted from certain fruits have been used as dyes in soft drinks, milk and powder products.  Anthocyanin is also used in the process of storing and preserving fruit, as well as making fruit jam.  The use of high-demand antosinanin is possible because it can reduce the use of synthetic dyes that are toxic and not environmentally friendly.  Anthocyanins are also used in making nutritional supplements because they have many positive effects on human health.  Flavonols are most commonly found as glycosides, usually 3-glycosides, and common flavonol aglycones are: Quercetin, Kaempferol, Myricetin  Fisetin as an antioxidant and anti- inflammatory  Quercetin can be found in various fruits, vegetables and leaves.  Quercetin can be used as a supplement, drink, or food ingredient.  Quercetin is an inhibitor of auxin transport to plants that appear naturally  In shallots, the largest concentration of quercetin is in the outer tuber layer and in the part closest to the root Quersetin as hepatoprotektor  Flavonol (Quercetin, Kaempferol, Myricetin, Fisetin),  Flavones (Luteolin, Apigenin),  Flavanon (Hesperetin, Naringenin),  Flavonoligans (Sibilinin),  Flavans (Catechin, Epicatechin),  Isoflavon (Genistein, Daidzein),  Anthocynaidins (Cyanidin, Delphidin),  Aurones (Leptosidin, Aureusidin),  Leucoanthocyanidis (Terasacidin),  Neoflavonoids (Coutareagenin, Dalbergin), Chalcones.  Many studies have proven that flavones are compounds with extensive bioactivity.  Among them, luteolin and apigenin are compounds with anti-cancer and antimutagenic abilities in vitro and in vivo.  Luteolin has a vasodilatory effect on rat aortic chest and apigenin which also has vasodilation effects can suppress skin tumor growth. Vasodilation is the dilation of the diameter of the blood vessels which occurs when the muscles in the blood vessel wall relax  Flavonoid isolation is generally carried out by extraction method, namely by maceration or soxhlation Flavonoids are generally soluble in polar solvents, except free flavonoids such as flavones, and flavonols are more soluble in semi polar solvents.  The extraction process, for screening and isolation purposes, generally uses methanol or ethanol. These solvents are dissolving compounds ranging from less polar to polar  Each fraction obtained was evaporated, then flavonoids were tested.  To detect the presence of flavonoids in each fraction, it is done by dissolving a small amount of thick extract per fraction into ethanol.  solvent front Rf of component A = dA component B dS Less polar! dS dB Rf of component B = dB component A dS More polar! dA The Rf value is a decimal fraction, generally only origin reported to two decimal places - The color samples are easy to be seen and no need to use UV lamp to detect them 76  The method commonly used is the method DPPH (1,1 diphenyl-2 picrylhydrazyl). In this method flavonoid antioxidants (AH) react with DPPH free radicals in a way:  Donate hydrogen atoms, cause DPPH color change from purple turn yellow.  Color intensity is measured by a spectrophotometer at a wavelength of 517 nm antioxidant Violet ------------------> Yellow  The extract produced by vacuum rotary evaporator was diluted into 5 concentration variations, namely 250 ppm, 500 ppm, 1250 ppm, 2500 ppm and 5000 ppm The aim is to determine the antioxidant activity of flavonoids by making an IC curve 50  The extract was reacted with 0.1 mM DPPH with a volume ratio of 1: 1 and incubated for 30 minutes, absorbance was measured at a wavelength of 517 nm Example DPPH test with Quersetin standard or Vitamin C  Example result of DPPH Sample plant extract  Determination of IC 50 by making a regression equation between the percentage of DPPH free radical activity in extracts on 5 extract concentrations  The antioxidant activity of flavonoids can be calculated by the formula: Antioxidant activity= Abs control –Abs sample x 100% Abs control  Value of IC 50 (Inhibitory Concentration) Calculation that use to determine reducing activity of free radical is to find the IC 50 value This value illustrates the concentration of the test compound that can reduce free radicals by 50%. The IC50 value is obtained based on the calculation of the regression equation by plotting the concentration of the test solution and the percent (%) inhibition or reducing of DPPH as a parameter of antioxidant activity.  Sample Concentration (ppm) as X- axis and persen value of inhibition as ordinate (Y-axis). The results of the linear regression equation and the results of the analysis of IC 50 values obtained from the absorbance of sample extracts and quercetin. Y= 0,335x + 6,617 Y value substituted by 50 Result : 50-6,617 = 43,38/0,335 = 129,49  Example result of DPPH Test Larutan Uji Persamaan Regresi IC50 (ppm) A Y= 1,335 +6,693 32,44 (1) B Y=0,295 +2,485 161,06 (4) C Y= 0,314 + 11,076 123,96 (3) D Y= 0,510 +10,723 77,01 (2) Standar Y= 9,055 + 5,128 4,950 (1) kuersetin Molyneux (2004) states that a substance has antioxidant properties if the IC50 value is < 200 ppm, but with a weak category.  Prepare quersetin standard in the etanol with concentration 40, 60, 80, 100 dan 120 μg/mL  Quersetin standard pipette @ 0,5 ml then added extract 0,5 ml and aquadest 2,5 mL  Add NaNO2 5% 0,15 mL, let stand for long 6 menit  Add 0,3 mL AlCl3.6H2O, incubation for long 5 minutes in the dark place  Measure absorbance in wavelength 507 nm  Create standar quersetin with regresion- correlation. Result of flavonoid total state with μg/mL THANK YOU