Phytochemical Composition of Mangifera Indica (Mango) Fruit Peel Extract

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Phytochemical Composition of Mangifera Indica (Mango) Fruit Peel Extract

Phytochemicals are compounds that act as free radical scavengers to help eliminate the highly charged oxygen molecules that are byproducts of metabolized oxygen (Khalid, 2007), and are believed to offer various health benefits (Van Duyn and Pivonka, 2000; Min et al., 2013). Medicinal plants are of great importance to the health of individual and communities.

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The medicinal value of these plants in some chemical active substances that produce a definite physiological action on the human body. The most important of these chemically active (bioactive) constituents of plants are: alkaloids, tannin, flavonoid and phenolic compounds. Many of these indigenous medicinal plants are also used for medicinal purposes (Edoga, 2005).
Mangoes belong to the genus Mangifera of the family Anacardiaceae. Mango trees grow up to 35– 40M (11-130ft.) tall with a radius of 10m (33ft).The trees are long leafed, as some specimens still fruit after 300 years. In deep soil the taproot descends to a depth of 6m (20ft), with profuse, wide spreading feeder roots; the tree also sends down many anchor roots, which penetrate several feet of soil. The leaves are evergreen alternate, simple, 15- 35cm (5.9-14 in) broad; when the leaves are young they are orange pink, rapidly changing to a dark, glossy red, then dark green as they mature. There are over 1000 named mango varieties trough out the world, which is a testament to their value to human kind. Mango is a common garden tree throughout the tropics when ripe, this delicious desert fruit is particularly high in Vitamin A. The fruit is also, an important source of sustenance for birds, bats, insects and mammals. Dried mango flowers, containing 15% tannin serve as astringents in cases of diarrhea, chronic dysentery and chronic urethrithis. The bark contains mangiferine and is astringent when used against rheumatism and diphtheria, leaf decoction when taken as a remedy for fever, chest pain, diarrhea, diabetes and hypertension. Extracts of bark, leaves, stem and unripe fruits are used as antibiotics for many ills. The plant is found abundantly across Nsukka, in the South-Eastern region of Nigeria. However, not much is known about the chemical composition of the plant leaves.

AIM: the aim of the present study was to determine the phytochemical constituents of Mangifera inidica.
OBJECTIVE: To extract crude extracts from mango fruit.
1. To determine the various phytochemicals, present in the extracts such as; alkaloids, tannins, glycosides, flavonoids, terpenoids, phenols, soluble carbohydrate, reducing sugar, saponins, steroids and hydrogen cyanide.

Mangifera indica
Mangoes belong to genus Mangifera which consists of about 30 species of tropical fruiting trees in the flowering plant family Anacardiaceae. According to ayurveda, varied medicinal properties are attributed to different parts of mango tree (Scartezzini and Speroni 2000).

Scientific Classification
Kingdom: Plantae
Division: Angiosperms
Class: Eudicots
Subclass: Rosids
Order: Sapindales
Family: Anacardiaceae
Genus: Mangifera
Species: Mangifera indica (Nunez et al., 2002).

Common names:
Sanskrit: Ambrah; Madhuulii; Madhuula; Madhuulaka; English: Mango; Igbo: mangoro; Hindi: Aam; French: mangot; mangue; manguier; Portuguese: manga; mangueira; Dutch: manja; Tamil: Ambiram; Mambazham; Mambalam; Mangai; Punjabi: Amb; Wawashi; Gujarati: Ambo, Keri; Marvo (unripe); Kashmiri: Amb; Malayalam: Amram; Choothaphalam; Manga; Manpalam; Mavu; Marathi: Amchur; Amba (Pott et al., 2003).


Botanical description
Mangifera indica is a large evergreen tree in the anacardiaceae family that grows to a height of 10-45 m, dome shaped with dense foliage, typically heavy branched from a stout trunk. It is native tropical Asia and has been cultivated in the Indian subcontinent for over 4000 years and is now found naturalized in most tropical countries. The leaves are spirally arranged on branches, linear-oblong, lanceolate – elliptical, pointed at both ends, the leaf blades mostly about 25-cm long and 8-cm wide, sometimes much larger, reddish and thinly flaccid when first formed and release an aromatic odour when crushed (Chen et al., 2004).

The inflorescence occurs in panicles consisting of about 3000 tiny whitish-red or yellowish – green flowers. The fruit is a well known large drupe, but shows a great variation in shape and size. It contains a thick yellow pulp, single seed and thick yellowish – red skin when ripe. The seed is solitary, ovoid or oblong, encased in a hard, compressed fibrous endocarp. Mango is one of the most popular of all tropical fruits. Mangiferin, being a polyphenolic antioxidant and a glucosyl xanthone, it has strong antioxidant, anti-lipid peroxidation, immunomodulation, cardiotonic, hypotensive, wound healing, antidegenerative and antidiabetic activities. Mangifera indica (MI), also known as mango, aam, it has been an important herb in the Ayurvedic and indigenous medical systems for over 4000 years. Various parts of plant are used as a dentrifrice, antiseptic, astringent, diaphoretic, stomachic, vermifuge, tonic, laxative and diuretic and to treat diarrhea, dysentery, anaemia, asthma, bronchitis, cough, hypertension, insomnia, rheumatism, toothache, leucorrhoea, haemorrhage and piles. All parts are used to treat abscesses, broken horn, rabid dog or jackal bite, tumour, snakebite, stings, datura poisoning, heat stroke, miscarriage, anthrax, blisters, wounds in the mouth, tympanitis, colic, diarrhea, glossitis, indigestion, bacillosis, bloody dysentery, liver disorders, excessive urination, tetanus and asthma. Ripe mango fruit is considered to be invigorating and freshening (Subha et al., 2007).

The juice is restorative tonic and used in heat stroke. The seeds are used in asthma and as an astringent. Fumes from the burning leaves are inhaled for relief from hiccups and affections of the throat. The bark is astringent, it is used in diphtheria and rheumatism, and it is believed to possess a tonic action on mucus membrane. The gum is used in dressings for cracked feet and for scabies. It is also considered anti-syphilitic. The kernels are converted into flour after soaking in water and eliminating the astringent principles. Most parts of the tree are used medicinally and the bark also contains tannins, which are used for the purpose of dyeing (Ornelas-Paz et al., 2007).
Chemical Constituents of Mango
Chemical constituents of MI are always of an interest. The different chemical constituents of the plant, especially the polyphenolics, flavonoids, triterpenoids. Mangiferin a xanthone glycoside major bio-active constituent, isomangiferin, tannins & gallic acid derivatives. The bark is reported to contain protocatechic acid, catechin, mangiferin, alanine, glycine, γ-aminobutyric acid, kinic acid, shikimic acid and the tetracyclic triterpenoids cycloart-24-en-3β,26diol, 3-ketodammar-24 (E)- en-20S,26-diol, C-24 epimers of cycloart-25 en 3β,24,27-triol and cycloartan-3β,24,27-triol. Indicoside A and B, manghopanal, mangoleanone, friedelin, cycloartan-3β-30-diol and derivatives, mangsterol, manglupenone, mangocoumarin, n-tetacosane, n-heneicosane, n-triacontane and mangiferolic acid methyl ester and others isolated from stem bark of MI. Mangostin, 29-hydroxy mangiferonic acid and mangiferin have been isolated from the stem bark together with common flavonoids. The flower yielded alkyl gallates such as gallic acid, ethyl gallate, methyl gallate, n-propyl gallate, n-pentyl gallate, n-octyl gallate, 4-phenyl gallate, 6-phenyl-n-hexyl gallate and dihydrogallic acid. Root of mango contains the chromones, 3-hydroxy-2-(4’-methylbenzoyl)-chromone and 3-methoxy-2-(4’-methyl benzoyl)-chromone. The leaf and flower yield an essential oil containing humulene, elemene, ocimene, linalool, nerol and many others. The fruit pulp contains vitamins A and C, β-carotene and xanthophylls. An unusual fatty acid, cis-9, cis-15-octadecadienoic acid was isolated from the pulp lipids of mango. Phenolic Antioxidants, Free Sugars and Polyols isolated and analyzed from Mango (MI) Stem Bark. All structures were elucidated by ES-MS and NMR spectroscopic methods. Quantitative analysis of the compounds has been performed by HPLC, and mangiferin was found to be the predominant component (Seifried et al., 2007).

Polyphenols have been characterized in mango puree concentrate by HPLC with diode array and mass spectrometric detection. A rapid method was developed for quantitative determination of beta-carotene, including cis-isomers, in dried mango. HPLC method was developed to determine carotenoids in Taiwanese mango. 5-Alkyl- and 5-alkenylresorcinols, as well as their hydroxylated derivatives, extracted from mango (MI) peels, purified on polyamide and characterized by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (HPLC/APcI-MS) for the first time. Xanthophyll esters, carotenes, and tocopherols has been identified and quantified in the fruit of seven mexican mango cultivars by liquid chromatography-atmospheric pressure chemical ionization-time-offlight mass spectrometry [LC-(APcI (+))-MS]. A simple, precise, and rapid HPTLC method was established for quantitative determination of the bioactive marker compound mangiferin in the stem bark & leaves of MI. The method was validated for selectivity, linearity, precision, accuracy, and robustness. The natural C-glucoside xanthone mangiferin [2-C-β-Dgluco-pyranosyl-1,3,6,7-tetrahydroxyxanthone; C19H18O11; Mw, 422.35; melting point, anhydrous 271°C has been reported in various parts of MI leaves, fruits, stem bark, heartwood and roots. The presence of a phenolic compound from leaves of MI which was named as homomangifirin (Pardo-Andreu et al., 2006).

Although a lot of pharmacological investigations have been carried out based on the ingredients present but a lot more can still be explored, exploited and utilized. Reactive oxygen species (ROS) possess a strong oxidizing effect and induce damage to biological molecules, including proteins, lipids and DNA, with concomitant changes in their structure and function. The major nutritional antioxidants, vitamin E, vitamin C and β-carotene, may be beneficial to prevent several chronic disorders considerable interest has arisen in the possible reinforcement of antioxidant defenses, both for chemoprevention and treatment purposes. The extract showed a powerful scavenging activity of hydroxy radicals and acted as a chelator of iron. It also showed a significant inhibitory effect on the peroxidation of rat brain phospholipid and prevented DNA damage caused by bleomycin or copper-phenenthroline systems. The interaction of Vimang (MI extract) with Fe (III) was studied and the results justify the high efficiency of Vimang as an agent protecting from iron-induced oxidative damage. The work has been carried out to investigate the pulp composition of four mango cultivars (Haden, Tommy Atkins and Ubá) at the ripening stage in relation to three components with antioxidant potential (total phenolics, carotenoids and ascorbic acid). These results corroborated previous information that mangoes are a good source of antioxidants in human diet. In vitro antioxidant and free radical scavenging properties of a stem bark aqueous extract of mango tree (MI), whose formulations are used in Cuba as food supplements under the brand name of Vimang, Luminol-enhanced chemiluminescence was used to elucidate the effect of this extract on the generation of reactive oxygen species in PMA- or zymosan-stimulated human polymorphonuclear leukocytes and on superoxide radicals generated in the hypoxanthine–xanthine oxidase reaction. Part of this MI extract antioxidant activity could be ascribed to the presence of mangiferin as its main component. The iron-complexing ability of Vimang as a primary mechanism for protection of rat liver mitochondria against Fe2+- citrate-induced lipoperoxidation was reported. The results are of pharmacological relevance since. Vimang could be a potential candidate for antioxidant therapy in diseases related to abnormal intracellular iron distribution or iron overload. The protective abilities of MI stem bark extract (Vimang ) 50-250 mgkg(-1), mangiferin 50 mgkg(-1) and selected antioxidants (vitamin C 100 mgkg(-1), vitamin E 100 mgkg (-1)and beta -carotene 50 mgkg(-1)) against the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced oxidative damage in serum, liver, brain as well as in the hyper-production of reactive oxygen species (ROS) by peritoneal macrophages was compared (Bidla et al., 2004).

A 50% ethanolic extract of the leaves of MI produced a significant hypoglycemic effect at a dose of 250 mg/kg, both in normal and streptozotocin-induced diabetic animals. The stimulation of β-cells to release insulin was thought to be part of the mechanism of action. The effect of the aqueous extract of the leaves of MI on blood glucose level in normoglycaemic, glucose – induced hyperglycaemic and streptozotocin (STZ)-induced diabetic rats has been assessed. The results indicate that the aqueous extract of the leaves of MI possess hypoglycaemic activity. This action may be due to an intestinal reduction of the absorption of glucose. The leaves of MI used for antidiabetic properties using normoglycaemic, glucose-induced hyperglycaemia and streptozotocin (STZ) induced diabetic mice. The aqueous extract of the leaves of MI possess hypoglycaemic activity. The effect of mango (MI) ingestion on blood glucose levels of normal and diabetic rats has been studied. The results from this research suggest that mango flour can possibly help in the treatment of diabetes (Rocha R et al., 2008).

Antiviral activity
In vitro the effect of mangiferin was studied against Herpes simplex virus type 2; mangiferin does not directly inactivate HSV-2 but inhibits the late event in HSV-2 replication. In vitro mangiferin was also able to inhibit HSV-1 virus replication within cells and to antagonize the cytopathic effects of HIV (Pardo et al., 2005).

Anthelmintic and anti-allergenic activity
Anthelminthic and antiallergic activities of MI stem bark components Vimang and mangiferin was investigated in mice experimentally infected with nematodes, Trichinella spiralis. The study was carried out to find out anti-allergic properties of vimang and mangiferin, a C-glucosylxanthone isolated from extract of MI. The results constitute the anti-allergic properties of Vimang on allergic models, as well as suggesting that this natural extract could be successfully used in the treatment of allergic disorders. Mangiferin, the major compound of Vimang, contributes to the anti-allergic effects of the extract (Andreas et al., 2000).

Antiparasitic activity
In a neonatal mouse model, mangiferin at 100 mg/kg has a similar inhibitory activity on Cryptosporidium parvum than the same dose (100 mg/kg) of an active drug, paromomycin (Aderibigbe et al., 2001).


Antibone resorption
Four water extracts of Kampo formulae were screened for their inhibitory effect on bone resorption induced by parathyroid hormone in organ culture of neonatal mouse parietal bones. Mangiferin isolated and tested in vitro showed a significant inhibitory effect on this model (Sanchez et al., 2000).

The significant cytotoxic activities has been demonstrated by the stem bark extract of mango against the breast cancer cell lines MCF 7, MDA-MB-435 and MDA-N, as well as against a colon cancer cell line (SW-620) and a renal cancer cell line (786-0). The ethanol/water (1:1) extract of dried aerial parts of mango administered intraperitoneally to mice at a dose of 250.0 mg/kg was inactive on Leuk-P388. In vitro, mangiferin dose- and time-dependently inhibited the proliferation of K562 leukemia cells and induced apoptosis in K563 cells line, probably through down-regulation of bcr/abl gene expression. These results suggest that mangiferin has a potential as a naturally-occurring chemopreventive agent (Amrita et al., 2009).

Antispasmodic and antipyretic activity
The stem bark extract of MI was evaluated for antiplasmodial activity against Plasmodium yoelii nigeriensis. The extract was also screened for antipyretic activity in mice. The extract exhibited a schizontocidal effect during early infection, and also demonstrated repository activity. A reduction in yeast-induced hyperpyrexia was also produced by the extract. The in vitro antimalarial activity of chloroform: methanol (1:1) extract of MI was evaluated. The extract showed a good activity on P. falciparum in vitro with a growth inhibition of 50.4% at 20 μg/mL (Muruganandan et al., 2005).

Immunomodulatory activity of alcoholic extract of stem bark of MI was investigated in mice. It is concluded that test extract is a promising drug with immunostimulant properties. Mangiferin mediates the down-regulation of NF-xB, suppresses NF-xB activation induced by inflammatory agents, including tumor nuclear factor (TNF), increases the intracellular glutathione (GSH) levels and potentiates chemotherapeutic agent-mediated cell death; this suggests a possible role in combination therapy for cancer. It is likely that these effects are mediated through mangiferin ROS quenching and GSH rising; increased intracellular (GSH) levels are indeed known to inhibit the TNF-induced activation of NF-κB (Garcia et al., 2003).

The potential anti-diarrhoeal activity of methanolic (MMI) and aqueous (AMI) extracts of seeds of MI has been evaluated in experimental diarrhoea, induced by castor oil and magnesium sulphate in mice. The results illustrate that the extracts of MI have significant anti-diarrhoeal activity and part of the activity of MMI may be attributed to its effect on intestinal transit (Rolo and Palmeira, 2006).

An ethanolic (95%) extract of the seed kernel of MI exhibited significant anti-inflammatory activity in acute, subacute and chronic cases of inflammation. The MI leaf extract exhibited antibacterial activity against Bacillus subtilis, staphylococcus albus and Vibrio cholerae. Analgesic and anti-inflammatory effects of MI extract (Vimang) has studied. The polyphenols found in the extract were found to account for the activity reported In vivo and in vitro anti-inflammatory activity of MI extracts (VIMANG) was investigated. MI extract, administered topically (0.5-2 mg per ear), reduced ear edema induced by arachidonic acid (AA) and phorbol myristate acetate (PMA, ED50 = 1.1 mg per ear) in mice. The results represent an important contribution to the elucidation of the mechanism involved in the anti-inflammatory and anti-nociceptive effects reported by the standard MI extract VIMANG (Perpétuo and Salgado, 2003)..

Anti-bacterial and antifungal activity
In an in vitro agar diffusion technique, mangiferin showed activity against 7 bacterial species, Bacillus pumilus, B. cereus, Staphylococcus aureus, S. citreus, Escherichia coli, Salmonella agona, Klebsiella pneumoniae, 1 yeast (Saccharomyces cerevisiae) and 4 fungi (Thermoascus aurantiacus, Trichoderma reesei, Aspergillus flavus and A. fumigatus) (Ojewole, 2005).

The antimicrobial activities of methanolic extracts of P. guajava and MI have been investigated. The results show that P. guajava and MI extracts exhibited antimicrobial activities at a concentration of 20 mg/ml. Overall, P. guajava extract show more antimicrobial activity than MI extract against tested organisms(Rivera et al., 2006).

Chemopreventive properties of lupeol and mango pulp extract (MPE) was evaluated against 7, 12- dimethylbenz (a) anthracene (DMBA) induced alteration in liver of Swiss albino mice. Lupeol/MPE was found to be effective in combating oxidative stress induced cellular injury of mouse liver by modulating cell-growth regulators (Yoshimi et al., 2001).

A novel gastroprotective agent, mangiferin, a naturally occurring glucosylxanthone from MI (Anacardiaceae), was evaluated in mice on gastric injury induced by ethanol and indomethacin. The effects of mangiferin on gastric mucosal damage were assessed by determination of changes in mean gastric lesion area or ulcer score in mice and on gastric secretory volume and total acidity in 4-h pylorus-ligated rats. These findings provide evidence that mangiferin affords gastroprotection against gastric injury induced by ethanol and indomethacin most possibly through the antisecretory and antioxidant mechanisms of action (Sarkar et al., 2004).

Other activity
Ethanolic extracts of Punica granatum, MI, Boerhaavia diffusa, Embelia ribes, Phyllanthus maderaspatensis, and Withania somnifera, has been tested for their effect on α-amylase activity (in vitro). P. granatum and MI were found to exhibit interesting α-amylase inhibitory activity. The ethanolic extracts of Lawsonia inermis leaves, Holarrhena antidysenterica bark, Swertia chirata whole plant and MI bark was tested for in-vitro α-glucosidase inhibitory activity. MI extract was found to be the most potent, with an IC50 value of 314 μg/ml. The effects of the MI (Vimang) extract, and mangiferin (a C-glucosylxanthone of Vimang) on the inducible isoforms of cyclooxygenase (cyclooxygenase-2) and nitric oxide synthase (iNOS) expression and on vasoconstrictor responses in vascular smooth muscle cells and mesenteric resistance arteries, has investigated respectively, from Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. They concluded that, the antiinflammatory action of Vimang would be related with the inhibition of iNOS and cyclooxygenase-2 expression, but not with its effect on vasoconstrictor responses. The activity of the MI leaf extracts against Clostridium tetani, has been investigated which causes many deaths around the world. Ether and ethanolic leaf extracts were obtained by sequential extractions (Perrucci et al., 2006).

The chemical tests showed that the ether extract had saponins, steroids and triterpenoids, while the ethanol extract had alkaloids, anthracenosides, coumarins, flavonones, reducing sugars, catechol and gallic tannins, saponins, steroids and triterpenoids. Both the ethereal and ethanolic fractions showed anti-clostridium tetani activity with an MIC of 6.25 and 12.5 mg ml–1, respectively (Sairam et al., 2003).

Phytochemicals are chemical compounds that occur naturally in plants (phyto means plant in Greek). Some are responsible for colour and other organoleptic properties, such as the deep purple of blueberries and the smell of garlic. The term is generally used to refer to those chemicals that may have biological significance, for example carotenoids or flavonoids, but are not established as essential nutrients (Palermo et al., 2014; Papp et al., 2007).There may be as many as 4,000 different phytochemicals having potential to affect diseases such as cancer, stroke or metabolic syndrome (Bongoniet al., 2013).

Tannin (also known as vegetable tannin, natural organic tannins, or sometimes tannoid a type of biomolecule, as opposed to modern synthetic tannin) is an astringent, bitter plant polyphenolic compound that binds to and precipitatesproteins and various other organic compounds including amino acids and alkaloids (Katie and Thorington, 2006).
The tannin compounds are widely distributed in many species of plants, where they play a role in protection from predation, and perhaps also as pesticides, and in plant growth regulation (McGee, 2004). The astringency from the tannins is what causes the dry and puckery feeling in the mouth following the consumption of unripened fruit or red wine. Likewise, the destruction or modification of tannins with time plays an important role in the ripening of fruit and the aging of wine (McGee, 2004).

Chemical Structure of Tannin

Phenol, also known as carbolic acid, is an aromaticorganic compound with the molecular formula C6H5OH. It is a white crystallinesolid that is volatile. The molecule consists of a phenyl group (-C6H5) bonded to a hydroxyl group (-OH). It is mildly acidic and requires careful handling due to its propensity to cause chemical burns (Kuttet al., 2008).
Phenol was first extracted from coal tar, but today is produced on a large scale (about 7 billion kg/year) from petroleum. It is an important industrial commodity as a precursor to many materials and useful compounds. Its major uses involve its conversion to plastics or related materials. Phenol and its chemical derivatives are key for building polycarbonates, epoxies, Bakelite, nylon, detergents, herbicides such as phenoxy herbicides and numerous pharmaceutical drugs (Kuttet al., 2008).

Uses of Phenol
The major uses of phenol, consuming two thirds of its production, involve its conversion to precursors to plastics. Condensation with acetone gives bisphenol-A, a key precursor to polycarbonates and epoxide resins (Pedro, 2009). Condensation of phenol, alkylphenols, or diphenols with formaldehyde gives phenolic resins, a famous example of which is Bakelite. Partial hydrogenation of phenol gives cyclohexanone, a precursor to nylon. Nonionic detergents are produced by alkylation of phenol to give the alkylphenols, e.g., nonylphenol, which are then subjected to ethoxylation (Pedro, 2009).
Phenol is also a versatile precursor to a large collection of drugs, most notably aspirin but also many herbicides and pharmaceutical drugs. Phenol is also used as an oral anesthetic/analgesic in products such as Chloraseptic or other brand name and generic equivalents, commonly used to temporarily treat pharyngitis (Pedro, 2009).

Flavonoids (or bioflavonoids) (from the Latin word flavus meaning yellow, their color in nature) are a class of plantsecondary metabolites. Flavonoids were referred to as Vitamin P (probably because of the effect they had on the permeability of vascular capillaries) from the mid-1930s to early 50s, but the term has since fallen out of use (Svobodovaet al., 2003). Chemically, they have the general structure of a 15-carbon skeleton, which consists of two phenyl rings (A and B) and heterocyclic ring (C). This carbon structure can be abbreviated C6-C3-C6. According to the IUPAC nomenclature (Svobodovaet al., 2003).

Functions of Flavonoid
Flavonoids are widely distributed in plants, fulfilling many functions. Flavonoids are the most important plant pigments for flower coloration, producing yellow or red/blue pigmentation in petals designed to attract pollinator animals (Galeottiet al., 2008). In higher plants, flavonoids are involved in UV filtration, symbiotic nitrogen fixation and floral pigmentation. They may also act as chemical messengers, physiological regulators, and cell cycle inhibitors. Flavonoids secreted by the root of their host plant help Rhizobia in the infection stage of their symbiotic relationship with legumes like peas, beans, clover, and soy (Galeottiet al., 2008). Rhizobia living in soil are able to sense the flavonoids and this triggers the secretion of Nod factors, which in turn are recognized by the host plant and can lead to root hair deformation and several cellular responses such as ion fluxes and the formation of a root nodule. In addition, some flavonoids have inhibitory activity against organisms that cause plant diseases, e.g. Fusariumoxysporum (Galeottiet al., 2008).

Alkaloids are a group of naturally occurring chemical compounds (natural products) that contain mostly basicnitrogen atoms. This group also includes some related compounds with neutral and even weakly acidic properties. Some synthetic compounds of similar structure are also termed alkaloids. In addition to carbon, hydrogen and nitrogen, alkaloids may also contain oxygen, sulfur and more rarely other elements such as chlorine, bromine, and phosphorus (Andreas, 2009).
Alkaloids are produced by a large variety of organisms including bacteria, fungi, plants, and animals. They can be purified from crude extracts of these organisms by acid-base extraction. Many alkaloids are toxic to other organisms (Andreas, 2009). They often have pharmacological effects and are used as medications, as recreational drugs, or in entheogenic rituals. Examples are the local anesthetic and stimulantcocaine, the psychedelic psilocin, the stimulant caffeine, nicotine, the analgesicmorphine (Raymond et al., 2010), the antibacterial berberine, the anticancer compound vincristine, the antihypertension agent reserpine, the cholinomimeticgalantamine, the anticholinergic agent atropine, the vasodilator vincamine, the antiarrhythmia compound quinidine, the antiasthma therapeutic ephedrine, and the antimalarial drugquinine (Raymond et al., 2010).
The boundary between alkaloids and other nitrogen-containing natural compounds is not clear-cut. Compounds like amino acidpeptides, proteins, nucleotides, nucleic acid, amines, and antibiotics are usually not called alkaloids. Natural compounds containing nitrogen in the exocyclic position (mescaline, serotonin, dopamine) are usually attributed to amines rather than alkaloids (Leland, 2006).

Biosynthesis of Alkaloids
Biological precursors of most alkaloids are amino acids, such as ornithine, lysine, phenylalanine, tyrosine, tryptophan, histidine, aspartic acid, and anthranilic acid (Blankenship et al., 2005). Nicotinic acid can be synthesized from tryptophan or aspartic acid. Ways of alkaloid biosynthesis are too numerous and cannot be easily classified. However, there are a few typical reactions involved in the biosynthesis of various classes of alkaloids, including synthesis of Schiff bases and Mannich reaction (Blankenship et al., 2005).

Synthesis of Schiff Bases
Schiff bases can be obtained by reacting amines with ketones or aldehydes. These reactions are a common method of producing C=N bonds (Faulkner et al., 2006).

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