Compounds from A. Platanoides Bark, V. Corymbosum Roots & Topical Formulations using Maple Syrup

The United States is the world’s largest producer of blueberries and the world’s second largest producer of maple syrup. Maine is the nation’s leading producer of wild blueberries, harvesting 82.6 million pounds of wild blueberries. Similarly, the New England region represents roughly 75% of the total U.S. production of maple syrup. These plants play a vital role in New England’s economy and are commonly consumed; however, these plants benefit more than the economy. The secondary metabolites of these plants are both antioxidants, and also anti-inflammatory, antibacterial and anti-viral agents. Blueberries are commonly consumed and have been investigated exhaustively. On the other hand, the roots and stems of the blueberry bush have not been investigated for their phytochemicals. Similarly, our laboratory has investigated two of the most common species tapped for their sap of the maple (Acer) genus. Another species of the maple family that produces sap is the Norway maple tree (Acer platanoides). Investigating the phytochemicals from these plants can revolutionize many industries.

x   there are a very large number of secondary metabolites yet to be discovered. 1 Living organisms produce organic compounds which can be broken down into primary metabolites and secondary metabolites. Primary metabolites, which include proteins, carbohydrates, fats, and vitamins, are compounds needed by an organism to exist. Secondary metabolites are compounds that are not directly involved in the normal growth, development or reproduction of organisms.

LIST OF TABLES
The term "phytochemical" refers to a wide variety of compounds in plants.
Phytochemicals are found in plant-based foods such as fruits, vegetables, beans and grains. Thousands of phytochemicals have been studied closely for their beneficial effects on human health.
Due to its global consumption, maple syrup has attracted scientific interest for a better understanding of its benefits beyond its sweet taste. Currently the sugar maple and red maple are the primary trees tapped for their sap which is boiled to produce syrup. However, as sugar maple (Acer saccharum) and red maple (Acer rubrum) have suffered in the northeastern United States due to an increase in climatic temperature, researchers have sought alternative sources of sap-producing trees. One such tree of interest is the Norway maple (Acer platanoides), known for its tolerance to higher temperatures and its ability to produce sap that can be concentrated to syrup.
Our laboratory recently has become interested in investigating the secondary metabolites of maple syrup and other plant parts of the most commonly tapped trees.
Our investigation of the phytochemicals of the sugar maple bark and the red maple stems and bark resulted in the isolation and identification of 15 novel compounds.

Acer saccharum: Sugar Maple
Sugar maple is the most abundant of the maple species found in New England.
Its economic importance in the production of maple syrup is one of the reasons why it is deemed the State Tree of New York. 5 The sugar maple is easy to identify by the clear sap in the petiole.
As previously mentioned, the bark of the sugar maple was investigated for its phytochemicals, resulting in the isolation of four novel phenolic glycosides named saccharumosides. These saccharumosides had cytotoxic activity against human colon tumor cell lines (HCT-116 and Caco02); in contrast, they were non-cytotoxic against non-tumorigenic cell lines (CCD-8Co). 2

Acer rubrum: Red Maple
Similar to the sugar maple, the red maple is tapped for its sap, which is then concentrated into syrup. The State Tree of Rhode Island, the red maple is native to eastern North America and adapts to most soil types and site conditions.
Our investigation of the secondary metabolites of the red maple's bark and stems resulted in the isolation and identification of nine novel gallotannins named maplexins. These gallotannins had potent α-glucosidase inhibition. Additionally, this study also suggested that the position of the galloyl groups on the glucitol core correlated to the inhibition of α-glucosidase. Moreover, the three galloylated derivatives attached to different positions of the glucitol moiety were 10-20 fold more potent α-glucosidase inhibitors than the clinical drug, Acarbose. 3

Acer platanoides: Norway Maple
Norway maple trees are native to Tromsø, Norway and have also been cultivated in Western Europe and in North America. 6 It is an invasive plant grown as a street tree and shade tree and favored due to its tolerance of poor, compacted soils and urban pollution, the Norway maple is deciduous and can reach a height of 30 meters, with a trunk up to 1.5 meters in diameter. 7 The Norway maple tree can live up to 250 years and is commonly confused with the sugar maple tree.
Due to the isolation and identification of novel bioactive compounds from edible and nonedible parts of the Acer species, our laboratory was interested in investigating other species in hopes of finding additional novel compounds.

Isolates from Acer genera:
Maple syrup is the largest commercially available and consumed natural product that is obtained entirely from the sap of deciduous trees. The sugar maple tree is the most commonly tapped for its sap. Although the sap is not as sweet as that of the sugar maple, the red maple and Norway maple are trees that also produce sap. The popular consumption of maple syrup has generated interest in isolation and characterization of its phytochemical constituents. Our laboratory has investigated sugar maple bark and red maple stems and bark. In vitro studies suggest that the consumption of these isolated phytochemicals may have health benefits.
Fifteen phenolics were isolated from the bark of the sugar maple tree (See Table A). Four of these compounds were novel phenolic glycosides shown to be cytotoxic against human colon cancer cells while non-cytotoxic against nontumorigenic cell lines. However, more studies would need to be conducted as in vitro activity does not correlate to in vivo effects. 2 Thirteen gallic acid derivatives were isolated from the stems of the red maple tree (See Table B). Of those thirteen phenolics, five novel gallotannins named maplexins with very potent α-glucosidase activity were isolated. Gallotannins are gallic acid derivatives that are esterified to a carbohydrate core. The galloyl groups attached to the glucitol varied in number and location. This study suggests the number and position of galloyl groups and location are directly correlated to the biological activity of these compounds. Ginnalin A has been investigated for its cytotoxic effects and determined to be more effective than both B and C against human colon tumorigenic cells (HCT-116) and breast cancer cells (MCF-7). This evidence further compels the investigation of other Acer species for their bioactive compounds. Our investigation of the red maple stems prompted further investigation into the bark of the tree. This study resulted in the identification of six novel compounds and 11 known phenolics (See Table C). The three galloylated derivatives attached to positions 1,5-anhydro-glucitol were 10-20 fold more potent α-glucosidase inhibitors than the clinical drug, Acarbose. 3 This suggests the location and number of galloyl groups is correlated to the biological activity for the inhibition of α-glucosidase. Hardwoods from other Acer species have also been investigated for their phytochemicals. The stems of the Ussuri maple (Acer barbinerve) were investigated and resulted in the isolation and characterization of 13 compounds (See Table D). 9 Furthermore, the compounds isolated are ubiquitous and commonly found in higher plants. [10][11][12] Among these, methyl gallate and gallic acid were also previously isolated from red maple bark and stems. 3,4  Another Acer species investigated for its phytochemicals was the Nikko maple (Acer nikoense). The Nikko maple bark was investigated because it was used in traditional Japanese folk medicine for hepatic disorders. The bark has also been investigated for anticancer, anti-inflammatory, antifungal and antibacterial effects.
Two novel compounds, acerogenin A and acerogenin B, along with nine known phenolic compounds were isolated and characterized (See Table E). These were evaluated for their role as inhibitors of Na+-glucose cotransporter. Na+-glucose cotransporter is a protein membrane that plays an important role in the reabsorption of glucose in the kidneys. 13 Additionally, the compounds isolated from the Nikko maple were evaluated for their anti-inflammatory effects. All compounds, with the exception of aceroside I, inhibited inflammation. Acerogenin A and acerogenin B moderately inhibited SGLT1. SLGT1 is expressed in the small intestine and responsible for the reabsorption of glucose. 13 It is believed the inhibition of SGLT could decrease the reabsorption of glucose resulting in an increase in urinary sugar excretion which may benefit type 2 diabetes patients.
The biological activity of these compounds from the different Acer species suggests investigating other species for novel phytochemicals that may have health benefits. The leaves of the Norway maple were previously investigated but only resulted in the identification of three compounds (See Table F). 14 One of the three compounds, cyanidin 3-(2",3"-digalloyl-β-glucopyranoside), was novel while the other two, cyanidin 3-(2"-galloyl-β-glucopyranoside) and cyanidin 3-β-glucopyranoside, have been previously reported. Thus investigating the bark of the Norway maple tree is essential since it is possible that these compounds can be re-isolated as well as novel compounds may be obtained. 14 Compounds Isolated from Norway Maple Leaves cyanidin 3-β-glucopyranoside 3-(2"-galloyl-β-glucopyranoside) cyanidin 3-(2",3"-digalloyl-β-glucopyranoside) Our laboratory has investigated several different species from the Acer genus. [2][3][4][15][16][17] The barks of many trees have been investigated, resulting in the isolation and characterization of many novel compounds. One of the most well-known anticancer compounds, taxol, was isolated from the bark of the Pacific yew tree. Taxol is one example of many phytochemicals that are used in medicine; its benefits establish a firm basis to investigate all parts of plants rather than only their edible parts.
Understanding the differences between compounds from different species of the Acer genus may yield insight into the chemotaxonomic distribution of compounds.
Additionally, it could explain which compounds are organ-specific since research has indicated that plants commonly contain organ-specific distributions of secondary metabolites. 18 Therefore, phytochemical isolation efforts on various plant parts have potential for the discovery of novel bioactive compounds.

Maple Syrup:
The province of Quebec, Canada is the largest producer of maple syrup and is responsible for about three-quarters of the world's output. Vermont is the largest producer in the United States, producing approximately 5% of the world's maple syrup.
Maple syrup is graded based on its viscosity and color. Sucrose is the most prevalent sugar in maple syrup accounting for over 61% of maple syrup's makeup. To produce maple syrup, sap must first be collected and boiled down 40x to obtain pure syrup.
Understanding the phytochemicals of maple syrup may lead to knowledge of additional benefits beyond its sweet taste. Our laboratory has extensively investigated maple syrup for its constituents, and evaluated its constituents for their biological activity. This investigation resulted in the identification of more than 70 compounds, some of which have potent antioxidant activity (See Table G). [15][16][17] Maple syrup's constituents, including high sugar content and a rich polyphenolic makeup, make it ideal for a topical formulation. The anti-inflammatory activity of the polyphenols and the high-sugar concentration could work synergistically as an anti-aging and moisturizing formulation. Vaccinium is a genus of shrubs in the Ericaceae family. The fruit of many different species are available commercially and widely consumed. These berries include the cranberry, blueberry, bilberry, whortleberry, lingonberry, cowberry and huckleberry. 19 The genus contains about 450 species and lives in habitats such as bogs and acidic woodlands. The plant structure differs between species as some lay low to the ground such as dwarf shrubs and some are larger shrubs growing up to 12-feet tall. 19 The fruits are usually berries and are red or bluish with purple juice.

Vaccinium corymbosum: Highbush Blueberry
A bush native to the United States, the highbush blueberry can grow up to 12feet tall. 19 The highbush blueberry gets its name from its tall stature compared to the low-lying, lowbush blueberry. Its twigs are yellow-green most of the year, but turn dark red in winter months. Its leaves are deciduous, alternate, simple, elliptic or ovate and slightly waxy on top. The white or pink flowers are small and urn-shaped with five petals and each cluster contains approximately eight to ten flowers. Flowering of the highbush blueberry occurs from February to June, and fruiting occurs from April to October. 19 The highbush blueberry is widespread in eastern North America. The most common native habitat is found in moist or wet soil of moderate to high acidity such as in marshes, swamps and lakes. Nonetheless, the bush also can be found in dry areas, such as dunes, barrier beaches and rocky hillsides. 19 The highbush blueberry produces fruit every year and is self-fertile. Crosspollination increases fruit set and results in larger and earlier berries with more seeds.
Bees are the primary pollinators. However, the seeds may be dispersed by birds and mammals. 19 In the southern portion of its range, highbush blueberry seeds have thick seed coats and require cold stratification before germination. Those from northern regions produce thinner seed coats and germinate in the autumn after dispersal. Plants have also been noted to produce root sprouts which emerge up to two meters away from the parent plant. 20

Phytochemicals Isolated from Highbush Blueberry Fruit:
Small berries constitute one of the important sources of potential healthpromoting phytochemicals. 21,22 These fruits are rich sources of phenolic compounds such as phenolic acids, anthocyanins, proanthocyanidins and other flavonoids, which have been investigated for their potential health-promoting effects. 21,22 The content of phenolics in berries is affected by the degree of maturity at harvest, cultivar, environmental conditions, storage conditions and processing. Polyphenols, abundant in blueberries, have been seen to produce favorable nootropic, antioxidant and antiinflammatory effects. [20][21][22] Many in vitro and in vivo studies suggest that the flavonoid-rich fruits of the blueberries (Vaccinium spp.) have the ability to inhibit and prevent diseases. The fruits contain a variety of phytochemicals, including anthocyanins, flavonols, proanthocyanidins, stilbenes and triterpenes. These phytochemicals could contribute to human health benefits. Blueberry constituents are antioxidants that counter oxidative stress and decrease inflammation.
Due to their superior antioxidant potential, the anthocyanins found in blueberries may play a major role in the inhibition of oxidative processes linked to diseases and cancer. Blueberries are rich in anthocyanins, including petunidin, malvidin, delphinidin, peonidin and cyanidin. 23 Understanding the phytochemical constituents of these plants may help in the elucidation of their health benefits.
Moreover, secondary metabolites are known to be organ specific, as parts of the plant produce different secondary metabolites. This organ-specificity justifies the need to investigate both edible and non-edible parts of plants. Although non-edible plant parts are not consumed, secondary metabolites can be isolated from these organs and used in dietary supplements.
As previously mentioned, investigating the phytochemicals of different plants and plant parts provides a better understanding of the plants' benefits to human health.
Zheng et al have isolated and quantified the major secondary metabolites found within the blueberry fruit (See Table H). Additionally, Wang et al isolated gallic acid, syringic acid, protocatechuic acid, β-sitosterol, ursolic acid and β-sitosterol-β-Dglucoside. 24 Other polyphenols identified include flavonol glycosides and catechins.

Antioxidant Activity of the Highbush Blueberry:
Antioxidant compounds in food play an important role in preventive health.
Evidence suggests that antioxidants reduce the risk for chronic diseases including cancer and heart disease. 18,20,22,24,[27][28][29] Primary sources of naturally occurring antioxidants are whole grains, fruits and vegetables. Antioxidants like vitamin C, vitamin E, phenolic acids and phytoestrogens have been recognized as having the potential to decrease the risk of disease. Most of the antioxidant compounds in a typical diet are derived from plant sources and belong to various classes of compounds with a variety of physical and chemical properties.
Blueberries are known for their high oxidant radical absorbance capacity (ORAC) activity to reverse age-related deficits in cognitive function. 29 Blueberries, spinach and strawberries were chosen for a study to determine the ability of short-term dietary supplementation to reverse age-related deficits in motor and cognitive functions. Blueberries were the most effective of the three foods to show signs of reversing age-related deficits in behavioral activities of rats. 29 Moreover, another study conducted at the University of California concluded a diet rich in foods and beverages containing flavonoids may decrease the risk of developing atherosclerosis, due to the ability of these compounds to inhibit lowdensity lipoprotein (LDL) oxidation and platelet aggregation. Therefore, phytochemicals present in antioxidant-rich foods may be beneficial in reversing the effects of behavioral aging. 25

Anticancer Activity of the Highbush Blueberry:
Berry extracts were evaluated for their ability to inhibit the growth of human oral (KB, CAL-27), breast (MCF-7) and colon (HT-29, HCT116) tumor cell lines at different concentrations. As the concentration of berry extract increased, the inhibition of cell proliferation also increased in all tumorigenic cell lines. 22 This suggests that the phytochemicals work synergistically.
However, independent phytochemicals can be assayed for their biological activity. Ursolic acid isolated from the highbush blueberry has been assayed against several cell lines; and, research suggests that it is a potent cytotoxic agent in human leukemia cells, lymphocytic leukemia cells p-388, human lung carcinoma cell A-549, and human colon (HCT-8) and mammary tumor cells (MCF-7). 24 This further suggests blueberries contain additional benefits beyond their basic nutritional value.

α-Glucosidase Inhibitory Activity of the Highbush Blueberry:
Recently the highbush blueberry has been investigated for its inhibition of αglucosidase. In comparison to Acarbose, all blueberry extracts showed similar αglucosidase inhibition capabilities. Preliminary studies with the Vaccinium genus showed that a proanthocyanidin (PAC) rich fraction had a lower IC 50 value than an anthocyanin-rich fraction, suggesting a PAC-rich extract is more beneficial for the inhibition of α-glucosidase. 20 Again, in vitro studies have focused on identifying the phytochemical constituents responsible for the observed anticancer and α-glucosidase inhibitory activity of these fruits. The highbush blueberry fruits were shown to inhibit α-amylase and α-glucosidase. Blueberry fruits and flowers have been researched for their phytochemicals; 18 however, the roots have not been investigated. Previously published research supports the need for researching these roots for other possible bioactive compounds that potentially may be used medicinally or in nutraceuticals.
Because secondary metabolites are formed through specific biosynthetic pathways, understanding these pathways is important. One of the most common pathways is the shikimate pathway.

Shikimate Pathway:
Investigating biosynthetic pathways is important because investigating the metabolic pathways of natural products allows for a fundamental understanding of the origin of secondary metabolites. There are many different pathways such as the shikimate pathway, mevalonate pathway and the acetate pathway. 30 The shikimate pathway is one of the most common metabolic pathways and is responsible for the formation of phenylpropanoids, coumarins and even phenolic acids (See Figures 1 and   2). The shikimate pathway is found in bacteria, fungi and plants. Phenylalanine and tryptophan are essential amino acids that are consumed while tyrosine is directly derived from phenylalanine. Phenylalanine, tyrosine and tryptophan represent a majority of the precursors for nearly the entire output of aromatic biosynthesis. 31 In higher plants on the other hand, the amino acids are the precursors for many secondary metabolites with aromatic ring structures. 31 Phenylalanine and tyrosine form the basis of the C6-C3 units found in many natural products including coumarins, lignans and flavonoids. Formulations are developed based on desired purpose for treating conditions such as acne, eczema, burn healing, anti-aging, etc. 32 The profile will include: the intended therapeutic indication, the preferred dosage form (cream, gel, ointment, etc.), the anticipated product strength (% active ingredient), the desired release profile with skin penetration goals, target shelf life and the desired cosmetic/aesthetic properties. 32 The formulation must allow for optimal penetration of the active ingredient into the skin. Skin pH is approximately 5.5. Thus the pH of the formulation may change following application to the skin. Many factors contribute to an optimal aesthetic and effective formulation. The aesthetic appeal of the formulation is just as important to the customer as its intended use, and an especially useful tool when attempting to differentiate oneself from one's competitors.
Healing a wound is a very complex process with many steps. Anti-aging, moisturizing and even antibacterial formulations need to meet a lot of the same criteria. A wound-healing formulation, on the other hand, must take all of those factors into consideration, and thus would make a great product for other uses.
In this respect, particular interest has been shown in natural products such as honey, due to its healing properties exhibited in vitro. 33 One study of honey in vitro showed extraordinary wound-healing activity. Investigated for its wound-healing properties, honey's powerful healing abilities have been attributed to its sugar content as a result of hyaluronic acid's composition of polysaccharides. These chains are made from modifications of the monosaccharide glucose. 21 Flavonoid-rich plants have been investigated for their wound-healing ability and are known to expedite wound healing. 12,22 Thus, high sugar content in conjunction with a high polyphenolic profile may expedite wound healing by reducing inflammation and increasing collagen stimulation.

Vitamins in Wound Healing:
Vitamins such as vitamin A (retinol), C (ascorbic acid) and E (α-tocopherol) have been investigated extensively for their beneficial effects in topical applications. 34,35 Research has found that vitamins C and E can help protect the skin against sun damage. It also has been theorized that they may reverse discoloration and wrinkles associated with aging and overexposure to the sun. These powerful antioxidants stabilize free radicals in order to slow the damage produced by harmful UVA-UVB rays. However, vitamins have never been used in conjunction with maple syrup. Fatsoluble and water-soluble applications have also been of interest due to the capabilities of dermal penetration in vivo.

Vitamins Assisting with Proper Wound Closure:
Compounds with high antioxidant activity such as vitamin E and vitamin C are commonly used post-surgery for treating scarring and assisting in wound healing. 5 Our laboratory has investigated the phytochemicals isolated from maple syrup that show promising antioxidant activity compared to vitamin C. 33,36 These phytochemicals' potent antioxidant activity supports the use of maple syrup for its anti-inflammatory effects and ultimately for topical applications in wound healing.
Moreover, Bartlett et al, 1942, have shown that a sufficient depletion of Vitamin C produces a decreased tensile strength in healing skin wounds, even when supplemented orally and intravenously. 34 Vitamin A (retinol) has proven to be effective when supplemented in wounds as exhibited by Seifter et al. 37 The importance of vitamin concentrations in a formulation extends beyond texture and aesthetic appeal. It is directly related to irritation when applied topically and creates adverse effects including a rash or even more severely a burn. 10  Furthermore, between 7-10% of maple syrup is added to the formulation to maintain optimal consistency and texture when applied. With a low concentration of maple syrup, vitamin concentrations will also be low and therefore will not contain an effective dose.

Role of Sugar in Wound Healing:
Sugar content has been shown to increase skin elasticity thereby helping with  Abstract: This is the first report of epi-catechin aldehyde, assigned the common name of aceraldehyde, being isolated and characterized from the bark of the Norway maple tree (Acer platanoides) along with 19 known phenolic compounds. All compounds were elucidated on the basis of spectroscopic analysis including 1D and 2D NMR data and compared to published NMR when available. These compounds were evaluated for their α-glucosidase inhibitory activity.

Introduction:
The barks of many trees have been investigated resulting in the isolation and characterization of many novel compounds. One of the most well-known anticancer compounds, taxol, was isolated from the bark of the Pacific yew tree.
However, it is not feasible to isolate large quantities of this compound from the bark of this tree, therefore taxol has been synthesized and is now commonly used as a chemotherapeutic agent. This establishes a firm basis to investigate all parts of plants rather than only edible parts.
Moreover, our laboratory has previously investigated the sugar maple's bark and the red maple's bark and stems. [1][2][3] The phytochemical investigations of the hardwoods of the sugar maple tree and red maple tree resulted in the isolation of: four novel phenolic glycosides from the sugar maple bark; and nine novel gallotannins and two new phenolic glycosides from red maple bark and stems. These novel compounds show promising bioactivity. [1][2][3] Maplexins A-E were isolated from red maple stems and examined for their αglucosidase inhibitory activity. The maplexins that contained two galloyl groups were more effective α-glucosidase inhibitors than those with only one galloyl group. This preliminary study suggested that number and location of galloyl groups correlated with the inhibitory activity of these compounds. Maplexins F, G, H and I, isolated from the red maple bark, were potent α-glucosidase inhibitors with up to 20-fold more potent inhibition compared to the control, Acarbose. 2 This study then provided additional evidence confirming that the number of galloyl groups and relative locations of these groups on the glucitol core affect the inhibitory activity of these compounds. 1 Additionally, crude extracts of red maple tissues such as branches, wood of branches, bark of branches, stem bark and whole twigs were evaluated for their antioxidant capacity. The study, designed to investigate natural antioxidants, indicated the stem bark had the most potent antioxidant capacity. The bark of branches also displayed antioxidant activity, albeit less active than the stem bark. 4 Saccharumosides are phenolic glycosides isolated from the sugar maple. These phenolics also showed promising biological activity. They were cytotoxic against human colon tumorigenic (HCT-116 and Caco-2) cells while non-cytotoxic against non-tumorigenic cell lines. 3 Hardwoods from other Acer species have also been investigated for their phytochemicals. The stems of the Ussuri maple (Acer barbinerve) were investigated and resulted in the isolation and characterization of 13 compounds. 5 Furthermore, the compounds isolated are ubiquitous and commonly found in higher plants. [6][7][8] Methyl gallate and gallic acid were also previously isolated from red maple bark and stems. 1,2 Another Acer species investigated for its phytochemicals was the Nikko maple (Acer nikoense). Two novel compounds, acerogenin A and acerogenin B were isolated and characterized. They were also evaluated for their role as inhibitors of the Na+glucose cotransporter, which plays an important role in the reabsorption of glucose in the small intestine. 9 Additionally, the compounds isolated from the Nikko Maple were evaluated for their anti-inflammatory effects. Acerogenin A, aceroside B and aceroside IV had the most potent activity.

The Nikko maple bark was investigated because it was used in traditional
Japanese folk medicine for hepatic disorders. The bark has also been investigated for its anticancer, anti-inflammatory, antifungal, and antibacterial effects. The biological activity of these compounds isolated from the Acer genus invokes investigation of other species from this genus for novel phytochemicals that may have biological properties.
The red maple and sugar maple did not have any compounds in common.
However, compounds isolated from the red maple hardwoods including stems and barks contained gallotannins named maplexins A-I and novel phenolic glycosides named rubrumosides A and B. The sugar maple also contained phenolic glycosides named saccharumosides A-D. Previous investigation of these hardwoods has prompted investigation of other barks within the same genus to compare phytochemicals between species belonging to the same genus. One such tree is the Norway maple tree, native to Tromsø, Norway, and cultivated in Western Europe and in North America. 10 The Norway maple is grown as a shade tree and favored due to its tolerance of pollution. To date, there is no published literature on the bark of this tree. The investigation of its leaves has resulted in the isolation of three compounds, cyanidin 3β-glucopyranoside, 3-(2"-galloyl-β-glucopyranoside) and cyanidin 3-(2",3"-digalloylβ-glucopyranoside). However, to our knowledge, no phytochemical investigation has been conducted on the hardwood of this species.     The biosynthesis of compound 19 is being proposed as a nucleophilic substitution where methyltransferase attacks C-8 and is methylated by Sadenosylmethionine (SAM) (See figure 7). This is then enzymatically oxidized twice by cytochrome p-450 yielding the aldehyde moiety at carbon 8b. This is initiated by Lmethionine being converted to SAM resulting in positively charged sulfur to initiate the nucleophilic substitution reaction. Subsequently, the methyl-building unit is introduced by a nucleophilic substitution reaction to begin methylation. This reaction requires a nucleophilic carbon; and, thus a phenol with an adjacent carbonyl group is susceptible to methylation.
In conclusion, this investigation of compounds isolated from Norway maple bark yielded 19 phenolic compounds. After a thorough phytochemical investigation was conducted, compounds were compared to other phytochemicals that previously had been isolated from the hardwood of other species of the Acer genus. These results are consistent with previously reported literature reporting phenolics to be the primary phytochemicals from the Acer genus. Additionally, secondary metabolites are known not only to be organ specific, they can also be genus specific. Many of the phenolic compounds such as gallic acid, 8 vanillic acid, 23 scopoletin, 19, 23 catechin 20 and other compounds are commonly found in many higher plants. These compounds have been investigated for their biological activity as free radical scavengers. 8,20,23,24 Furthermore, another compound isolated from other Acer species is cleomiscosin C. It is a coumarolignan that has been isolated from the hardwood of trees including the Sugar maple and the Korean maple (Acer okamotoanum). 6,25 Cleomiscosin C has also been previously isolated from the stem wood of Aquilaria agallocha, and seeds of Cleome viscoas. 6 This coumarolignan has also been synthesized using fraxetin as a starter unit, 21 and shown to have significant antioxidant potential. 25,26 This provides evidence that compounds may be organ specific. Moreover, this also suggests these non-edible plant parts should be investigated for their phytochemicals, specifically those that, due to potential health benefits, could be used in nutraceutical formulations.

platanoides) Bark
Raed Omar, Tao  This manuscript is written to be submitted to Journal of Natural Products ACS.
Abstract: Highbush blueberry fruits and flowers were shown to inhibit α-amylase and α-glucosidase enzymes, relevant to type-2 diabetes management. Blueberry fruits and flowers have also been investigated for their phytochemicals. However, the roots have not yet been investigated. Here we report the first isolation and elucidation of cinnamtannins B1 and D1 isolated from the highbush blueberry roots along with three other known phenolic compounds.

KEYWORDS: Vaccinium corymbosum, highbush blueberry, roots, phenolics
Introduction: Roots play a very important role for a plant. They are responsible for the plant's absorption of minerals, oxygen and water. In winter months, the roots reserve food needed by the tree to produce spring foliage. 1 They are also responsible for anchoring the plant/tree above ground. Therefore, it is important to keep the portion above ground healthy by ensuring adequate water and rich soil for the roots to continue their functions. However, because the roots are exposed to many different environmental conditions to which the rest of the plant is not, they most likely produce different phytochemicals compared to the other organs of the plant. By maintaining healthy roots, the plant is able to produce better quality products such as berries.
Berry fruits are widely consumed in our diet and have attracted much attention due to health benefits for which they have been investigated extensively. Berries contain a diverse range of phytochemicals with biological properties which act as antioxidants, and anticancer and anti-inflammatory agents. [2][3][4][5][6][7][8][9][10] Flavonols have been of particular interest as they are known antioxidants agents that may inhibit the onset of coronary heart disease. 11 The phenolic content in berries is affected by the degree of maturity at harvest, cultivar, environmental conditions, storage conditions and processing. Polyphenols, abundant in blueberries, have been seen to produce favorable nootropic, antioxidant and anti-inflammatory effects. One of the most popularly consumed berries is the blueberry. Blueberries are consumed for reasons beyond their taste. Research has suggested blueberries have many preventative properties. The fruits, leaves and flowers are rich sources of vitamins A and C, and nutrients such as flavonoids and anthocyanins. 9 Furthermore, highbush blueberry flowers and fruits were shown to inhibit α-amylase and α-glucosidase enzyme activity, relevant to type-2 diabetes management. 3,7,10 Phytochemical isolation efforts on various plant parts reveal their potential to contain novel compounds. Many studies have been conducted on the berries and flowers of the highbush blueberry. However, the roots have not been investigated. 3,7 The objectives of this project were to isolate and elucidate secondary metabolites in highbush blueberry roots. This is the first phytochemical and biological study of roots of the highbush blueberry species.  Sunnyvale, CA, USA) and compared to that of the control, which had 50 µL buffer solutions instead of the test compounds. The α-glucosidase inhibitory activity was expressed as percent inhibition and was calculated as follows:

Identification of Compounds:
The isolated compounds (chemical structures shown in Figure 1     This manuscript is written to be submitted to World Academy of Science, Engineering and Technology

Introduction:
An emulsion is a preparation of two immiscible substances. Creams are an example of a common emulsion with a water phase and an oil phase. For these phases to be made miscible, an emulsifying agent is employed (i.e. bees wax). A water in-oil emulsion is commonly used for the treatment of dry skin. Active ingredients with specific biological properties such as antibacterial, antiviral, and anti-inflammatory properties, may enhance a formulation for a desired cosmetic effect. Cosmetic formulations are commonly enhanced with antioxidants.
Honey has been investigated for its wound-healing properties; and, its powerful healing abilities have been attributed to its sugar content. Flavonoid-rich plants have been researched for their wound-healing ability and anti-aging potential. 1,2 An emulsion with high sugar content in conjunction with a high polyphenolic profile may assist with transepidermal water loss contributing to dry skin. Honey and maple syrup have similar phytochemical constituents composed primarily of sugar. [3][4][5][6] The phytochemicals of maple syrup have been investigated along with vitamin and sugar analysis. Maple syrup, like honey, is composed primarily of sugar. Sugar content has been shown to assist with increasing skin elasticity to help with proper wound closure. 7-9 Maple syrup's combination of phytochemicals and high sugar content suggests it can act as a potent anti-aging moisturizing formulation.
Moreover, Cooper et al reported that a minimum of 29% sugar content is required for inhibition of Staphylococcus aureus. 10 Furthermore, they reported better inhibition at higher concentrations, compared to the lower concentrations of honey.
Sugar may play a role in inhibiting bacterial growth.
Hyaluronic acid (HA) consists of disaccharide chains made from modifications of the monosaccharide glucose. 11 It is believed that glucose derived from sugar may be converted into hyaluronic acid at the wound surface, forming an extracellular matrix. 9 HA is thought to aid in dermis healing. It is believed to assist in activating the inflammatory response, promoting cell proliferation, migration, and promoting reepithelization. 12 Therefore, HA may assist in moisturizing in topical formulation.
Due to the increased awareness of the adverse effects of synthetic compounds, an investigation of the moisture retentive properties of a maple syrup based emulsion was conducted compared to a base without maple syrup. Maple syrup is natural and free of any coloring or additives. It is boiled down directly from tree sap, which is harvested from the maple tree towards the end of winter. Maple syrup is not processed and contains vitamins and minerals, including calcium, potassium, sodium and copper suggesting it may be a candidate for an active ingredient as part of a moisturizing formula. 5,13 General Experimental Procedures: 100% Pure Maple Syrup was purchased from A&P in Saddle Brook, New Jersey. A digital pH-Meter by General Tools & Instruments Company LLC.

Preparation of Water-to-Oil Emulsion
In this study, a water-to-oil emulsion was prepared by the addition of aqueous phase to the oily phase with continuous agitation. Oily phase consisted of paraffin oil (16%) and surfactant ABIL-EM 90 (4%) heated up to 75°C. Water soluble ingredients were heated to 75°C and the maple syrup extract (12%) was then added to the experimental cream. The aqueous solution was added to the oily phase while stirring was continued at 3,500 rpm by the high-shear mixer for about 15 minutes until complete aqueous phase was added. After the complete addition of the aqueous phase, the speed of the mixer was reduced to 1,500 rpm for homogenization, for a period of 5 minutes, and the emulsion was then left to cool at room temperature. The base was prepared using the same method without the addition of the maple syrup.

Physical analysis of formulations:
The emulsion was analyzed based on color, thickness, look, feel and emulsion properties.

pH determination
pH value of freshly prepared emulsion and emulsions kept at different conditions were determined by a digital pH-Meter by General Tools & Instruments Company LLC.

Stability tests
Stability tests for the emulsions were performed in several different conditions. These storage conditions were analyzed to see whether they had an effect on the emulsion.
Tests were performed on samples kept at 8°C (in a refrigerator), 25°C (in incubator), 40°C (in incubator), 40°C (in incubator) with 45% relative humidity and 40°C (in incubator) with 75% relative humidity. Physical characteristics were also monitored daily. Photo-oxidation was also tested via exposure to UV rays 100-400 nm in a dark box for one month and monitored for physicochemical changes. formulations were evaluated at 8°C, 25°C, 40°C and 40°C and 75%RH. Stability was achieved after three weeks stored in variable conditions. The physical appearance and smell of the experimental maple syrup formulation and base formulation were not altered. There was no crystallization observed within the container or after application to the skin. Information about the emulsion and the temperature's effect on the formulation, the emulsion underwent temperature cycles from 5 to 40 °C within 24 h for four weeks. This is frequently used in industry as an accelerated stability test to predict long-term stability. The pH of the base and maple syrup were 5.9 and 5.2 respectively. The newly formulated maple syrup cream is suitable for improved skin-hydration levels. By reducing trans-epidermal water loss in people with dry skin, and decreasing the appearance of wrinkles, maple syrup cream acts a humectant. Previous literature supports the hypothesis of topical use of a high-sugar polyphenolic-rich active ingredient to increase moisture retention and wound healing, or as a potent antioxidant. 7 Antioxidants protect human skin from free radicals produced by UV radiation; and, flavonoids are known to be potent antioxidants and photo-protective agents.
Additionally, trans-epidermal water loss results in loss of collagen regeneration, 14 and the loss of collagen causes wrinkles.
The benefits of topically applied sugar have been investigated and show improvements on tensile strengthening. Moreover, sugar's antibacterial capabilities make it ideal when used with potent antioxidants for anti-aging capabilities. 1,2 Moreover, vitamins including as vitamin A (retinol), 15 vitamin E (αtocopherol), and Vitamin C (ascorbic acid) are known to assist in wound healing.
Topical application of vitamin C has been shown to elevate cutaneous levels of vitamin C, which correlates with the antioxidant protection of the skin protecting the skin from UVB damage. 16 Additionally, vitamin C was observed to play an important role increasing type I and type III collagen levels. 17 Vitamin C has been shown to be a potent anti-oxidant removing dead skin cells with vitamins A and E, the formulation assists by removing age spots and healing wounds, ultimately rejuvenating the skin.