If I were an apple,and grew on a tree......

  seminal trace...Apple Polyphenols.Apple Proanthocyanidins,Apple Proanthocyanidin B2,.M.F.Molecular Formula:C30H26O12.Procyanidin B-2, Extracted from Apples,Apple Procyanidins,Polyphenol from Apple,gallic acid,flavan-3ols,flavonols,phloridzin, cinnamates, hydroxymethyl furfural and anthocyanins,Apple Polyphenol Extracts....

 

   If I Were An Apple

    If I were an apple
  And grew on a tree
      I think I'd drop down
    On a nice boy like me.
        I wouldn't stay there
      Giving nobody joy
      I'd fall down at once
    And say, "Eat me, my boy


   Basic Botanical Info of Apple:

   Botanical: Pyrus malus
   Synonyms:Wild Apple. Malus communis.
   Parts Used:Apple peel and Apple core
   Habitat:Temperate regions of the Northern Hemisphere.
   Apples contain a large concentration of flavonoids, as well as a variety of other phytochemicals, and the concentration of these phytochemicals may depend on many factors, such as cultivar of the apple, harvest and storage of the apples, and processing of the apples. The concentration of phytochemicals also varies greatly between the apple peels and the apple flesh.
   In numerous epidemiological studies, apples have been associated with a decreased risk of chronic diseases such as cardiovascular disease, cancer, and asthma.
   


 Basic Botanical Info of Apple:
 From Findings of Apple Procyanidin B-2:
 Properties and effects of Polyphenol from Apple:
 Processing Method:Concentration of phenolics extracted from apples:
 Apple Polyphenol and Its Application to Tooth coating composition:
 Phloridzin-rich phenolic fraction and use thereof as a cosmetic, dietary or nutraceutical agent:
 Research and Findings:Apple Polyphenol
 Apple Polyphenol:toxicology and safety.
 How search engine think about Apple:
 Research update of Apple and Apple Phytochemicals:


   From Findings of Apple Procyanidin B-2:

  Procyanidin B-2, Extracted from Apples, Grows Hair in Clinical Trials:

 Late in 2000,Researchers presented their findings to the international community on the hair growing effects of apple polyphenols- specifically one known as procyanidin B-2.
 The researchers had spent years testing over 1,000 botanical extracts for hair growth properties. They identified two successful compounds- one from chardonnay grapes, and one extracted from unripe apples.
 A year before the landmark Marburg conference, tests were conducted on mouse and human skin cells.

 Here is what the cell culture tests revealed:

 

 The procyanidin B-2 fraction clearly outperformed the grape extract, and nearly doubled the hair growth activity of minoxidil. "Procyanidin B-2 purified from apples," stated the research team, "shows the highest activity of more than 300% relative to controls.":
 Promising results, indeed. Next came the in vivo mouse studies. Lab mice were shaved, then treated with procyanidin B-2, procyanidin C-1 (also from apples), minoxidil, and placebo. Results of these tests showed "extensive hair growth" for both procyanidins B-2 and C-1, as well as minoxidil.

  Investigation of topical application of procyanidin B-2 from apple to identify its potential use as a hair growing agent, Phytomedicine, 2000:

 In a double-blind placebo-controlled trial, nineteen men with male pattern baldness were studied with a daily topical application of a 1% procyanidin B-2 solution, extracted from apples. Ten other balding men served as controls, receiving a placebo solution. After 6 months, the study concluded:

 The increase in number of total hairs and terminal hairs in the procyanidin B-2 group subjects was significantly greater than controls
 78.9% of subjects showed an increased mean value of hair diameter
 "Procyanidin B-2 therapy shows promise as a cure for male pattern baldness."

  Procyanidin B-2, extracted from apples, promotes hair growth: a laboratory study, Br J Dermatol. 2002 Jan;146(1):41-51

 In this study, the researchers concluded that procyanidin B-2 acts to diminish protein kinase C isozymes, which play an important role in the hair growth cycle.
 Procyanidin B-2 seems to promote hair growth by downregulating PKC in both the anagen (active growth phase) and telogen (resting phase) of the hair follicle. When the anagen phase is prolonged, and the telogen phase is shortened, increased hair growth results.

 Two more clinical trials and a total of seven published studies have now confirmed the surprising hair growth-promoting effects of apple procyanidins. Here is a summary of those findings:
  Total Number of Hairs: Significantly Increased
  Total Number of Terminal Hairs: Significantly Greater
  Increase in Hair Diameter: 78.9% Positive
  Ratio of Thicker (terminal) Hairs: Significantly Higher
  Compared to Minoxidil: Up to 200% Better
  Hair Follicle Activation: Intensive

 

  Here are key findings:

 1. Multiple Procyanidins Produce Synergistic Results:

 First, a careful review of the data revealed that both procyanidin B-2 and procyanidin C-1 had profound effects on hair growth, but at different phases of the hair growth cycle. Further, the procyanidin B-3 fraction also showed significant results: even better than procyanidin B-2 in one testing phase.
 The first improvement to the formula was delivered- by enriching it with all three procyanidin oligomers- B-2, B-3 and C1.

 2. Apple Procyanidins are Dose-dependent in Nearly Every Study:

 The team then turned their attention to the concentration of the lab formula. Was a 1% solution- the amount used in the clinical trials- really the best possible strength for stimulating new hair growth?
 Based on months of cross-discipline research review of apple procyanidins and polyphenols, another clear trend emerged. In nearly every area of research, the effects of procyanidins were dose-dependent.
 When researchers treated breast cancer tumors with apple procyanidins, the effects were markedly enhanced at higher concentrations. In tests on colon cancer, skin cancer, Alzheimer's, and heart disease, the results were always the same: the higher the concentration, the better the results.
 Since procyanidins are thoroughly tested safe, and produce zero irritation or unwanted side effects, there seemed to be no reason to stop at a measly one percent. So they quadrupled the procyanidin strength of the new formula.
 


   Properties and effects of Polyphenol from Apple:

  1).  Suppresses the production of the melanin.
  2).  Suppresses the functioning of tyrosine enzymes.
  3).  Reliable results for whitening.
  4).  Used as an external cosmetic with apple polyphenol has the double,effect of protecting the skin and alleviating allergic dermatitis.
  5).  Prevents free radicals and ultraviolet ray damage to the skin.
  6).  Reliable sun-proof effects.
  7).  Applied as a sun lotion.
 


   Processing Method:Concentration of phenolics extracted from apples:

 The present invention also relates to the process of obtaining products used as food additives having enlarged concentrations of phenolics, the process involving extraction of phenolics from fruits, particularly from apples.

  A:Normal composition of Apple Polyphenols:  gallic acid, flavan-3ols, flavonols, phloridzin, cinnamates, hydroxymethyl furfural and anthocyanins.

 It has been known in the prior art that certain compounds or ingredients, present in fruits, particularly in grapes and apples, possess various benefits for human health apart from the well known benefits of vitamins, minerals and other nutrients considered to be the main ingredients of fruits. The terms "phenolics" or "plant phenolics" have been developed and been adopted in the art for a class or type of these compounds because these compounds include in their formula a hydroxyl (OH) function attached to an aromatic carbon atom in analogy to the well known chemical phenol, or these compounds have a chemical structure closely related to or derived from a compound having an aromatic (phenolic) hydroxyl group. Usually plant phenolics have more than one aromatic (phenolic) hydroxyl group. Generally speaking, plant phenolics have antioxidant properties and have been shown in in vitro and in in vivo studies to have positive effects on the human cardio-thoracic condition. The phenolics isolated from fruits, primarily from apples, include gallic acid, flavan-3ols, flavonols, phloridzin, cinnamates, hydroxymethyl furfural and anthocyanins.

  B:Raw material and part used:

 Because of the richness of apples as a source of plant-phenolics and because of the availability of apple peel and core as a relatively economical raw material, efforts in accordance with the present invention to produce isolates containing high concentration of phenolics have been focused on apple peel and apple core serving as a source.
 

  C:Summary and Processing:

 It is an object of the present invention to obtain from fruits, and particularly from apples, an extract which is rich in natural phenolics of the fruit and which is utilized as a supplement or additive in various food products.

 It is another object of the present invention to obtain from fruits, and particularly from apples, an extract which is rich in natural phenolics of the fruit and which does not impart an unnatural or undesired taste component to the food product in which it is used. It is still another object of the present invention to provide food products such as beverages, dairy products, frozen products, jams, jellies, preserves, and confectionary products which have an enlarged concentration of phenolics that have been extracted from fruits, particularly from apples, in accordance with the present invention.

 The foregoing objects and advantages are attained by a product or products that is obtained by extracting the fruit, or parts of the fruit such as peels and or cores with hot water in a temperature range of approximately 65 to 93 Deg C. (150 to 200 Deg F.), followed by depectinization of the extract with a pectinase enzyme, preferably at an elevated temperature of approximately 38 to 57 deg C. (100 to 135 deg F.) until test taken for pectin is substantially negative. The pectinase enzyme is then deactivated by heating the mixture to approximately 60 to 93 deg C. (140 to 200 deg F.) and the aqueous solution/suspension is filtered to yield discardable solids and an aqueous filtrate. Polyvinylpolypyrrolidone (PVPP) adsorbent is then added to the filtrate aqueous solution and the mixture is agitated to extract by adsorption of the plant phenolics contained in the aqueous phase. The polyvinylpolypyrrolidone (PVPP) adsorbent containing adsorbed plant phenolics is then collected by filtration, centrifugation or like method. The aqueous filtrate at this stage is substantially devoid of recoverable plant phenolics but after concentration can still serve as useful product, primarily as a delivery vehicle for the plant phenolics obtained by the following additional process steps.

 The collected solid polyvinylpolypyrrolidone (PVPP) adsorbent containing adsorbed plant phenolics is then washed multiple times with water to remove solids other than the adsorbed plant phenolics. The plant phenolics are obtained from the PVPP adsorbent by treating the adsorbent with dilute sodium hydroxide (or other acceptable strong base) solution, to give an aqueous extract of the plant phenolics, having a concentration of approximately 400 to 5000 mg gallic acid equivalent per liter (GAE/L) of the extract. The polyvinylpolypyrrolidone (PVPP) adsorbent recovered by filtration can be regenerated for repeated use by careful washing with water until all base is substantially removed from it.

 The aqueous extract containing the plant phenolics can be further concentrated to give a liquid product with greater concentration of plant phenolics. Alternatively the aqueous extract is mixed with a nutritionally acceptable carrier, such as maltodextrin, rice dextrin, modified corn starch or other carrier, and the mixture is converted to a solid by spray drying or like drying process effective to yield a solid product. When in a solid form, such as the spray dried product obtained from the aqueous extract, the concentration of phenolics depends on the amount of carrier (if any) or carriers added and on the moisture content of the solid. Preferably the spray dried solid product contains approximately 10 mg or more gallic acid equivalent (GAE) per gram of solid (1% by weight). When the aqueous extract is concentrated but not dried completely, it typically contains approximately 50,000 to 200,000, preferably approximately 100,000 mg GAE/L.

 The liquid concentrate, dried solid product or their solutions can be added to diverse types of food products, such as beverages, dairy products, frozen products, jams, jellies, preserves, and confectionary products to provide or enrich the products with fruit phenolics.

 The features of the present invention can be best understood together with further objects and advantages by reference to the following description, taken in connection with the accompanying drawings.
 

  Measurement of Plant-Phenolic Content:

 The Folin-Ciocalteau method is used to measure the phenolic content of the products using gallic acid as a standard. Total phenolic content was determined colorimetrically, using a Beckman DU-50 spectrophotometer. The molecular weight of gallic acid is 170.1. A standard solution of gallic acid at 0.01 M is made by completely dissolving with stirring 1.701 grams of gallic acid in 1 liter distilled water. Diluted solutions of gallic acid in millimolar concentrations ranging from 0.00 mM to 0.2 mM are created as standards from distilled water and this standard solution. A sodium bicarbonate solution of 0.71 M strength is made by dissolving 59.64 grams of the substance in 1 liter of distilled water.

  Folin-Ciocalteau Assay:

 0.1 gram phenolic substance (plant extract) to be assayed is weighed out and diluted to 10 grams with water and further diluted with another 6 grams of 0.71 M sodium bicarbonate. This mixture is allowed to react with 0.1 gram Folin-Ciocalteau reagent for 2 hours. A colored complex develops during this reaction period. The colored reaction medium is then measured for absorbance at 660 nm using the spectrophotometer. By comparison with an absorbance versus gallic acid equivalent per liter (GAE/l) curve that has been obtained by measurement of the standard solutions (standard curve) the concentration of plant phenolics in the assayed product is obtained (expressed as mg Gallic Acid Equivalents (GAE)/Liter). The phenolic standard gallic acid, sodium bicarbonate, and Folin-Ciocalteau reagent were obtained from Sigma Chemical Company, St Louis, Mo.


   Apple Polyphenol and Its Application to Tooth coating composition:

 Accordingly, an object of the invention is to provide a tooth coating composition having a quick drying property as demanded by general consumers and being excellent in the durability while having the merits of tooth coating compositions of prior art.

 As the result of making intensive investigations in view of the above-described circumstances, the present inventors have found that by using specific mica titanium for a tooth coating composition using shellac as the main body, a tooth coating composition, which does not become white turbid for a long time, has a quick drying property, and is excellent in the durability, is obtained and have accomplished the present invention.

  Then, the present invention described below in detail:
 

 Shellac used in the invention is a resin, which has actual results of being used for foods and has a very high safety. In the tooth coating composition of the invention, the shellac is used as a coat-forming component. The content of such a shellac in the total composition is preferably in the range of from 1% by weight to 20% by weight. When the content of shellac is less than 1% by weight, the coat is too brittle and thus the composition cannot be used as a tooth coating composition. Also, when the content thereof exceeds 20% by weight, the viscosity of the coating liquid is increased, whereby it becomes difficult to thinly coat the composition on teeth and also the drying property becomes inferior. The particularly preferred content of shellac is from 5% by weight to 15% by weight.

 In addition to shellac as the coat-forming component, one kind or a combination of two or more kinds of other resins, high molecular materials, and dental cements can be added to the composition at a suitable amount. By adding such material(s), the preferred effects of more improving the luster of the coat and more improving the dispersion of mica titanium or pigments can be expected.
 Examples of such a resin include an acrylic resin, a vinyl acetate resin, an alkyd resin, a vinyl chloride resin, a silicone resin, a fluorine resin, rosin, etc.
 Examples of the high molecular material include vinylon, nylon, polyvinyl alcohol, polyvinyl pyrrolidone, polyvinyl butyral, polybutene, polyethylene, polypropylene, polyisoprene, cellulose ethers, nitrocellulose, ester gum, viscose rayon, polysaccharide, etc.
 

 Furthermore, examples of the dental cements include zinc phosphate cement, carboxylate cement, glass ionomer cement, zinc oxide euginol cement, silicate cement, etc.

 Mica titanium is a substance obtained by coating a mica powder with a thin film of titanium oxide, and fundamentally has been frequently used as a pigment but in the invention, by compounding mica titanium as a coat-strengthening agent, the tooth coating composition excellent in the durability after coating is obtained. Also, by controlling the thickness of the titanium oxide film of the mica titanium, it is possible to obtain an interference color such as yellow, red, blue, green, etc., whereby the tooth coating composition giving a good aesthetic appearance is obtained.

 The compounding amount of such a mica titanium in the whole composition is preferably from 0.5% by weight to 30% by weight. When the content of mica titanium is less than 0.5% by weight, the strength of the coat is lowered and the desired durability is not obtained. On the other hand, the content thereof exceeds 30% by weight, the dispersibility thereof in the solution becomes inferior, also, not only after coating, the evaporation of the solution is delayed to reduce the drying property, but also after coating, a rough-feeling sense of incongruity occurs in the mouth. Particularly preferred compounding amount of mica titanium is from 2% by weight to 15% by weight of the whole composition.
 The mean particle size of mica titanium used in the invention is in the range of preferably from 5 um to 50 um, and more preferably from 10 um to 30 um. By using the mica titanium having the mean particle size of from 5 um to 50 um, a proper hiding power is imparted to the coat and further the using feeling in the mouth is improved.

 As the main solvent of the tooth coating composition of the invention, a lower alcohol-base solvent dissolving shellac is used. As the solvent, which is nontoxic to the human body and dissolves well shellac, there are ethanol, n-propanol, isopropanol, n-butanol, isobutanol, etc., and from the points of the dissolving property and the volatile property, ethanol is most preferred. In addition to the main solvent, as an auxiliary solvent, any solvent, which is safe for the human body and gives less stimulation in the mouth, may be used. For example, as an auxiliary solvent, there are water, propylene glycol, glycerol, etc. Propylene glycol and glycerol are also frequently used as the solvents for general cosmetics and are used for dissolving medical components and extracts.
 To the tooth coating composition of the invention can be added a chemical having an anti-cariogenic action. As such a chemical having an anti-cariogenic action, there are sodium fluoride, sodium monofluorophosphate, stannous fluoride, and polyphenol-containing vegetable extract.

 There are actual results that sodium fluoride, sodium monofluorophosphate, or stannous fluoride is properly compounded in dentifrices as a medical component having an anti-cariogenic action and many products are on the market. Also, it is known that the polyphenol-containing vegetable extract has an antimicrobial action to the S. Mutans bacteria, which is a decayed tooth bacteria, and the extract is considered to be a very effective dental carries preventing agent.
 Practical examples of the polyphenol-containing vegetable extract include one or more extracts selected from the group consisting of a fig extract, a hydrangea tea extract, an oolong tea extract, a tea extract, a grape seed extract, a grape rind extract, a blue berry extract, an apple extract, a eucalyptus extract and a rosemary extract.

 By compounding the above-described medicine having the anti-cariogenic action with the tooth coating composition of the invention, without making a troublesome action such as teeth polishing such as brushing, etc., the dental caries preventing effect can be expected.

 Also, the tooth coating composition of the invention can, if necessary, contain various coloring agent. Examples of such a coloring agent include zinc oxide, titanium oxide, barium sulfate, mica, carbon black, white carbon, calcium hydrogenphosphate, calcium tertiary phosphate, hydroxy apatite, iron oxide, chromium oxide, magnesium oxide, calcium oxide, aluminum oxide, fish flakes, talc, calcium carbonate, magnesium carbonate, barium carbonate, and tar dyes.
 

 Furthermore, the tooth coating composition of the invention can contain, if necessary, various pharmaceutical components other than the above-described substances and also spicery, an antiseptic, a buffer, s pH controlling agent, a surface active agent, a dispersing agent, a plasticizer, a ultraviolet absorbent, a viscosity controlling agent, an antioxidant, etc.

 The pharmaceutical components include sodium azulenesulfonate, Epsilon-aminocaproic acid, allantoin, allantoin chlorohydroxy aluminum, allantoin dihydroxy aluminum, epidihydrocholestrin, dihydrocholesterol, sodium chloride, glycyrrhizinic acid, diammonium glycyrrhizinate, di-sodium glycyrrhizinate, tri-sodium glycyrrhizinate, di-potassium glycyrrhizinate, mono-ammonium glycyrrhizinate, beta-glycyrrhizinic acid, isopropyl methyl phenol, cetylpyridinium chloride, decalinium chloride, benzalkonium chloride, benzethonium chloride, alkyldiaminoethylglycine hydrochloride, chlorohexidine hydrochloride, tricrosane, ascorbic acid, sodium ascorbate, pyridoxine hydrochloride, dl-alpha-tocopherol acetate, dl-alpha-tocopherol nicotinate, zeolite, di-sodium dihydrogen pyrophosphate, sodium pyrophosphate, sodium hydrogenphosphate, sodium tertiary phosphate, sodium polyphosphate, polyethylene glycol, polyvinyl pyrrolidone, lysozyme chloride, copper chlorophyllin sodium, hinokitiol, polyoxyethylene lauryl ether, lauroylsarcosine sodium, etc.
 The above-described pharmaceutical components have the actual results of being compounded with dentifrices and are preferred materials as the materials of being used in the mouth. As the pharmaceutical effects thereof, there are, in addition of the prevention of dental caries, the prevention of halitosis, the prevention of the deposition of tartar, the prevention of gingivalise.gingivitis, the effect of removing nicotine, etc.

 In addition, as the dispersing agent described above, there is an N-methacrolylethyl-N,N-dimethylammoniume. alpha-N-methylcarboxybetainee.butyl methacrylate copolymer. The compound functions as a good dispersing agent.


   Phloridzin-rich phenolic fraction and use thereof as a cosmetic, dietary or nutraceutical agent:

 The present invention relates to a phenolic fraction from fruit and also to the process for obtaining this fraction. This extract rich in an antioxidant compound, phloridzin, can be used as a cosmetic, dietary or nutraceutical preparation.

 It is known that polyphenolic compounds are relatively widespread and in large amount in the plant kingdom. In the Rosaceae family in particular, the analysis of the polyphenols of apple has led to the identification of at least 37 phenolic compounds, the most abundant of which are chlorogenic acid, procyanidins B1 and B2, epicatechin, phloretin, phloridzin and p-coumaric acid. Some of these compounds have physiological properties such as antioxidant, anti-mutagenic, antiallergic, anticancer and antidiabetic properties, and the like.

 Among the phenolic compounds, phloretin and its glycosylated derivative, phloridzin, are typical of apple and the other fruit of the Rosaceae family. In particular, phloridzin is found in large amount in the pips, but it is also present in apple juice and skin. Phloridzin has antioxidant activity allowing a cardiovascular protection similar to that of estrogens. Moreover, phloridzin is capable of acting on melanogenesis by activating a cascade of enzymes including tyrosinase, thus allowing increased protection against ultraviolet radiation. Phloridzin also has antidiabetic action by competitive inhibition of the sodium-dependent blood transport of metabolites such as glucose, galactose and the like. Phloridzin is also involved in inhibiting the growth of tumor cells by blocking the activity of protein kinase C.
 

 However, in apple (homogenate or juice), the dihydrochalcones (phloretin and phloridzin) are present in small amount relative to the other polyphenols. Chlorogenic acid and the procyanidins are the major polyphenols in apples, whether these are cider apples or dessert apples, phloridzin and phloretin never representing more than 5% by weight of the total polyphenols of ripe cider apples (analysis of 15 different varieties).
 In the known polyphenolic extracts, the proportions between the various phenolic molecules are conserved relative to the proportions present in the various starting materials, with the exception of the polymeric procyanidins, which are lost or degraded during the extraction.
 Polyphenolic extracts rich in hydroxycinnamic acids (caffeic acid, chlorogenic acid and coumaric acid) and in flavanols (catechin, epicatechin and procyanidins), and poor in dihydrochalcones (phloridzin and phloretin) are thus conventionally obtained.

  Details of the invention as following:

 The polyphenolic fraction in accordance with the invention comprises at least 20% by weight of polyphenols and preferably 50%, at least 10% by weight of which is composed of phloridzin, and preferably between 10% and 70%. This extract can also comprise in its composition chlorogenic acid, epicatechin, procyanidin B2, quercitrin, p-coumaric acid and also phloretin.

 One composition that is particularly preferred of this polyphenolic fraction is that it contains, by weight: more than 20% and preferably more than 50% of total polyphenols; at least 30% and preferably from 30% to 40% by weight of the polyphenols as phloridzin; not more than 11% and preferably between 2% and 11% by weight of the polyphenols as chlorogenic acid; not more than 4% of the polyphenols as epicatechin; not more than 2% of the polyphenols as procyanidin B2; not more than 1.5% of the polyphenols as quercitrin; not more than 0.4% of the polyphenols as p-coumaric acid; and less than 0.2% of the polyphenols as caffeic acid.

 Another subject of the invention is that the caffeic acid is present in weight proportions of less than 20% of the weight of phloridzin present. Preferably, the caffeic acid represents less than 1% by weight of the total polyphenols in the extracts. The proportion of phloridzin is 9 times as great by weight as that of the catechin. The amount of phloridzin present is at least equivalent by weight to that of the chlorogenic acid.

 Another subject of the invention is characterized in that it contains phloretin. By means of a controlled acid hydrolysis, virtually all of the phloridzin can be converted into phloretin, which is less water-soluble.

 Another subject is characterized in that the dihydrochalcones are present in proportions of greater than or equal to 40% by weight relative to the hydroxycinnamic acids.

 The extraction process for selectively extracting a dihydrochalcone-rich polyphenolic fraction from ripe apples is characterized in that it comprises the following steps: the crushed apples are subjected to one or more solid/liquid extractions, in the presence or absence of added water; the wet solid extract obtained is then either dried or enzymatically liquefied to obtain a liquid extract; the dry solid extract undergoes further extractions over a period of between 10 minutes and 2 hours with a polar organic solvent, preferably a C₁ C₄ aliphatic alcohol, pure or as a mixture with water, to obtain an organic extract; this organic extract is evaporated to dryness at a temperature of less than or equal to 60 deg C., preferably under reduced pressure; this residue is then taken up in water, after which it is depleted several times, preferably 4 times, with a water-immiscible solvent, preferably ethyl acetate or methyl or propyl acetate; the organic solutions obtained are mixed together and evaporated to dryness at a temperature of less than 60 deg C., and preferably less than 50 deg C., to obtain the polyphenolic fraction which is the subject of the present invention.

 Via another route, the wet solid extract is mixed with water in the presence of an enzymatic mixture for a period of between 1 and 4 hours at a temperature of between 30 and 50 deg C., and preferably between 40 and 45 deg C., to obtain a liquid extract; this liquid extract is clarified by centrifugation or by filtration and then by ultrafiltration; the extract is loaded onto a chromatography column filled with an adsorbent resin of styrene-divinylbenzene type. The resin is washed with acidified water to remove the impurities and the residual sugars. The polyphenols are then eluted with an aqueous-alcoholic solution containing between 40% and 70% and preferably between 50% and 60% by weight of ethanol. Other C₁ C₄ aliphatic alcohols may be used, such as methanol or butanol; if necessary, a dewaxing step is introduced during the process; the product obtained by extraction is taken up a final time in water and then dried, preferably by atomization or lyophilization to give a beige-colored powder containing at least 20% by weight of polyphenols, preferably more than 50% of polyphenols, 10% by weight of which, and preferably between 10% and 70% of the polyphenols are dihydrochalcones, preferably phloridzin.

 The fractions are obtained by the process described above, preferably from ripe apples of the Rosaceae family and in particular of the species Malus sylvestris Mill.

 This extract of eating apples having the characteristics stated, obtained according to the process described above, may be used as a dietary or nutraceutical supplement.
 The dihydrochalcone-rich phenolic fraction according to the invention has properties as an agent for protecting against ultraviolet radiation, and has antioxidant properties.
 A subject of the invention is also a cosmetic composition comprising, inter alia, the dihydrochalcone-rich polyphenolic fraction described above.

 The compositions according to the invention may be ingested or applied to the skin. According to the mode of administration, the composition according to the invention may be in any form usually used in cosmetics.
 The compositions according to the invention may especially be in the form of gel capsules, milks, lotions, creams, gels or beverages.
 The nutraceutical, dietary or cosmetic compositions of the present invention are conventionally formulated according to the applications for which they are intended.
 The examples which follow illustrate the invention without limiting it in any way.
 


   Research and Findings:Apple Polyphenol

  Optimization of apple polyphenol oxidase inactivation by high hydrostatic pressure.:

 M. MIYAWAKI, J. I. Reyes-De-Corcuera, J. R. Powers, and B. G. Swanson. Dept. of Food Science and Human Nutrition, Washington State Univ., PO Box 646376, Pullman, WA 99164-6376

 High Hydrostatic Pressure (HHP) utilization for food processing offers advantages over conventional thermal processing improving retention of "fresh-like" natural flavors and essential nutrients. One of the research objectives of HHP processing is focused on inactivation of endogenous detrimental enzymes in plant foods. Polyphenol oxidase (PPO) in apples is partially inactivated by HHP following mechanisms dependent on tissue status and process conditions. Our objectives are to establish optimal conditions of HHP treatments combined with antibrowning agents to inactivate PPO in apples and minimize undesirable browning during storage, and to develop shelf stable foods containing apples. HHP treatments were carried out at 600 MPa for up to 10 min at ambient temperatures to simulate potential commercial applications, followed by PPO assays and color observations for 2 days at ambient temperature. Diced apples and apple homogenates were subjected to HHP treatments to compare the effects of tissue and cell disruption on PPO inactivation. The soluble solids concentrations selected to simulate apple products, as well as organic and inorganic antibrowning agents were evaluated to determine their effects on the discoloration of HHP treated diced apples and apple homogenates. HHP inactivates PPO in diced apples more effectively than in apple homogenates. However, after HHP treatment at 600 MPa for 5 min at ambient temperatures, diced apples exhibited 8% PPO activity and slight brown discoloration. The decrease in PPO inactivation in apple homogenates during HHP treatments as soluble solids concentration increased is attributed to baroprotective effects and reduced water activity. Among the antibrowning agents, ascorbic acid and calcium chloride inhibited discoloration of apple homogenates before and after HHP treatments. Complete inactivation of PPO in apple homogenates by HHP treatments alone is unlikely. Inactivation of PPO in apple homogenates containing selected antibrowning agents by HHP treatment will slow the development and extent of undesirable brown discoloration.
 

  Anti-allergic effect of apple polyphenol on patients with atopic dermatitis: A pilot study.:

 Abstract:The aim of the present study was to evaluate the anti-allergic effect of apple condensed tannins (ACT) in patients with atopic dermatitis (AD) as a pilot study. An ACT supplement given to the patients at oral doses of 10 mg/kg per day for 8 weeks reduced the inflammation, lichenification, cracking, itching, sleep dis- turbance and peripheral blood eosinophil counts. Itching and sleep disturbance scores after ACT supplement even for 2 weeks were significantly decreased compared with the control group. The results suggest that ACT has an anti-allergic effect and that its use improved the symptoms of AD.

  Research on Scavenging Effect on Radical and Inhibiting Effect on Lipid Peroxidation of Different Apple Polyphenol Extracts in vitro.:

 Objective: To study the scavenging effects on hydroxyl radical (-OH), superoxidized anion radical (O(superscript - subscript 2)?) and malondialdehyde (MDA) generated by lipidperoxidation of different apple polyphenol extracts. Methods: The DR spectrophotometry and improved TBA spectrophotometry and solvent box method were used to determined the scavenging effects on ?OH, O(superscript - subscript 2)? and MDA. Results: Different apple polyphenol extracts all had antioxidant effects, but in different antioxidant systems, different apple polyphenol extracts had different antioxidant effects, and the antioxidant effects of AP I arid AP II showed better. Conclusion: Apple polyphenol extracts had antioxidant activity. The main antioxidant components were phenols and proanthocyandins oligomer with low molecular weight.

  Apple Polyphenol Oxidase Inhibitor(s) From Common House Fly.:

 Interpretive Summary: Enzymatic browning associated with polyphenol oxidase (PPO) causes millions of dollars in annual losses to the fruit, vegetable and seafood (crustaceans) industries. The catalytic action of PPO is usually a limiting factor in the handling and technological processing of many fruits and vegetables. Few preservatives are available to the food processing industry and one group, the sulfites, may be harmful to asthma sufferers. Because PPO is known to exist in insects, scientists at the USDA Center for Medical, Agricultural, and Veterinary Entomology, Gainesville, FL, worked with scientists at the University of Florida, Department of Food Sciences and Human Nutrition, to determine the existence of PPO inhibitor(s) in the house fly, Musca domestica. Crude inhibitor(s) was isolated that inhibited the activity of apple PPO up to 90% at pH values greater than 5.0. The highest degree of PPO inhibition was found in isolates from third-instar larvae and 3- to 5-day-old pupae. The PPO inhibitor(s) is most stable at pH 5.0 and quite stable to heating, thawing, and irradiation. This discovery may be useful to the fruit, vegetable, and seafood industries as an alternative mathod for combating enzymatic browning.

 Technical Abstract: A new apple polyphenol oxidase (PPO) inhibitor(s) from house fly (Musca domestica L.) was discovered. Crude inhibitor(s) isolated by buffer extraction, heat treatment and dialysis from house fly pupae inhibited the activity of apple PPO up to 90% at pH values above 5.0. Inhibition was strictly pH dependent. Inhibitor(s) was further characterized by employing heat, freezing and thawing, irradiation, pH adjustment and ultrafiltration studies. The potential PPO inhibitor(s) was quite stable to heating at 100 C for 1h, repeated freezing and thawing, and irradiation. Inhibitor(s) was most stable at pH around 5.0 and least stable at alkaline pH. PPO inhibition profile of house fly during its metamorphosis was also evaluated.

  Prevention of bone loss with apple polyphenol.:

 Prevention of bone loss by phloridzin, an apple polyphenol, in ovariectomized rats under inflammation conditions. Puel C, Quintin A, Mathey J, Obled C, Davicco MJ, Lebecque P, Kati-Coulibaly S, Horcajada MN, Coxam V. Unite des Maladies Metaboliques et Micronutriments, INRA Theix, 63122, Saint Genes-Champanelle, France.

 Aging and sex hormones related changes lead to inflammatory and oxidant conditions, which are involved in the pathogenesis of osteoporosis. Recent studies have suggested that polyphenols may exert a protective effect in such conditions. We assessed the effect of phloridzin (Phlo), a flavonoid exclusively found in apple, on bone metabolism in ovariectomized (OVX) or sham-operated (SH) rats with and without inflammation. Six-month-old Wistar rats were allocated to two equal groups that received either a control diet or a diet supplemented with 0.25% Phlo for 80 days. Three weeks before necropsy, inflammation was induced by subcutaneous injection of talc in 10 animals of each group. At necropsy, ovariectomy decreased both total (T-BMD) and metaphyseal (M-BMD) femoral bone mineral density (P < 0.01). Inflammation conditions, checked by an increase in the spleen weight and alpha1-acid glycoprotein concentration in OVX rats, exacerbated the decrease in T-BMD (g/cm2) (as well as M-BMD) observed in castrated animals (P < 0.05). Daily Phlo intake prevented ovariectomy-induced bone loss in conditions of inflammation as shown by T-BMD and M-BMD (P < 0.05).

 At the diaphyseal site, BMD was improved by Phlo in OVX rats with or without inflammation (P < 0.05). These results could be explained by changes in bone remodeling as the increased urinary deoxypyridinoline excretion in OVX and OVXinf animals was prevented by the polyphenol-rich diet (P < 0.001), while plasma osteocalcin concentration was similar in all experimental groups. In conclusion, Phlo consumption may provide protection against ovariectomy-induced osteopenia under inflammation conditions by improving inflammation markers and bone resorption.
 

  Apple polyphenols lower blood fat.:

 Just last month, researchers at Cornell University announced that extracts from apples lowered "bad" (LDL) cholesterol in cell tests. Six prior studies in animals and humans also show a cholesterol lowering effect from apple polyphenols.
 A 2003 study on apple phytochemicals suggested "a strong link between dietary intake of phytochemicals and reduced risk of cardiovascular disease."

 Several studies show that apple polyphenols have a positive effect on diabetes and insulin resistance in animals and humans. In 1987, European researchers announced that an apple-derived polyphenol, phlorizin, "completely normalized insulin response" in diabetic rats.
 In 2004, scientists at the National Institutes of Health in the U.S. gave this same apple polyphenol to mice. Two weeks of treatment "significantly decreased blood glucose levels" in diabetic mice. Whole body fat mass was also "significantly reduced."
 An Asian study in diabetic human volunteers showed that even weak polyphenols in apple juice produced "avoidance of sharp peaks" in blood glucose levels.


   Apple Polyphenol:toxicology and safety.

  Safety and Acute toxicity:

 General Safety Status: Apple polyphenol extracts are listed ("apple essence, natural") in the FDA's approved additive/GRAS (Generally Recognized As Safe) database.Researchers found that apple polyphenol extracts are safe, even at concentrations 200 times the recommended human dosage.

 Acute toxicity(LD50):Procyanidin B-2 [epicatechin-(4beta --> 8)-epicatechin]

 LD50-Lethal dose,50 percent kill.Rat.Subcutaneous.2000 mg/kg. (subcutaneous injection)
 Primary irritation tests using rabbits indicated that procyanidin B-2 containing preparation shows no primary irritation. In the guinea pig maximization test, there was no evidence of sensitization to procyanidin B-2.In primary ocular irritation tests using rabbits, procyanidin B-2 containing preparation and vehicle showed slight irritation of conjunctivae which is assumed to be caused by ethanol. It is suggested that topical procyanidin B-2 is safe and acceptable from the series of toxicological tests.
 Reference:Food Chem Toxicol. 1999 May;37(5):545-52.PMID: 10456683.

 Longterm Use: Apple polyphenol extract has strong antioxidant activity and various physiological functions, and is used as a food additive and nutritional supplements. Here, we tested the consumption safety of Applephenon, which is a polyphenol extract produced from unripe apples. The Ames test without S9 mixture revealed that Applephenon, had slight mutagenicity at a high concentration of 2500 microg/plate; however, both chromosomal aberration test and the micronucleus test found no significant mutagenicity. Furthermore, an acute oral-toxicity test, and a 90-day subchronic-toxicity test showed no significant hematological, clinical, chemical, histopathological, or urinary effects at a dose of 2000 mg/kg. These results confirm that Applephenon is safe and no toxic at average dietary level.
 Reference:Food Chem Toxicol. 2004 Jun;42(6):959-67.PMID: 15110105.

 Toxicological studies on procyanidin B-2 for external application as a hair growing agent.:

 Procyanidin B-2 [epicatechin-(4beta --> 8)-epicatechin] is one of condensed tannin that exists widely in plants. We have reported previously that procyanidin B-2 possesses hair epithelial cell growth-promoting activity and stimulates anagen induction in hair cycle progression.
 To evaluate the safety of topical procyanidin B-2 as a hair growing agent, we examined the mutagenicity, acute subcutaneous injection, primary irritation, skin sensitization, and eye irritation of this compound. Mutagenicity tests using bacteria showed procyanidin B-2 to be non-mutagenic. Chromosomal aberration tests using CHL cells indicated that procyanidin B-2 caused polyploidy but no structural aberrations. In micronucleus tests for mutagenicity using mice, procyanidin B-2 was negative.
 Acute subcutaneous injection study using rats revealed no symptoms of significant injury. The lethal dose of procyanidin B-2 is greater than 2000 mg/kg (subcutaneous injection). Primary irritation tests using rabbits indicated that procyanidin B-2 containing preparation shows no primary irritation. In the guinea pig maximization test, there was no evidence of sensitization to procyanidin B-2.
 In primary ocular irritation tests using rabbits, procyanidin B-2 containing preparation and vehicle showed slight irritation of conjunctivae which is assumed to be caused by ethanol. It is suggested that topical procyanidin B-2 is safe and acceptable from the series of toxicological tests.
 Reference:Food Chem Toxicol. 1999 May;37(5):545-52.PMID: 10456683.


  Scientific References:

  1.If I were an apple,and grew on a tree.