CHAPTER I -Introduction
CHATER II -Review and Literature
2.1 History of MoringaOleifera.
2.2 Moringa Species
2.3 Classification MoringaOleifera
2.4 Common Name of Moringa Oleifera
2.5 Morphology and Physical Characteristics
2.8 Chemical Composition
2.11 Antibacterial Activity
2.13 Use of Moringaoleifera
CHAPTER-III MATERIAL AND METHOD
3.1 Experimental site
3.2 Experimental material
3.3 Glass wares and polywares
3.4 Chemicals and solvents
3.5 Biochemical parameter
3.6 Determination of phytochemicals
3.7 To record qualitative analysis of amino acid by TLC
3.8To assay antibacterial activity of differentextract of moringa
3.9 To find out mineral composition of different parts of moringa using XRF
3.10 Estimation of sodium by flame photometer
3.11 To study heavy metal in different parts of moringa using VA
CHAPTER-IV RESULT AND DISCUSSION
4.1 Biochemical parameter
4.2 Phytochemicals analysis
4.3 To record qualitative analysis of amino acid by TLC
4.4 To Assay antibacterial activity of different extract of moringa oleifera
4.5To find out mineral composition of different parts of moringa using XRF
4.6 Estimation of sodium by Flame Photometer
4.7 To study heavy metal in different parts of moringa using VA
The authors are thankful to the Department of Chemistry, KSKV Kachchh University-Bhuj and Department of Biotechnology, Junagadh Agricultural University-Junagadh for providing necessary facilities to carry out the research work.
LIST OF FIGURES
1.1 Different use of moringa oleifera
2.1 Moringa oleifera
2.2 Trunk and leaves of moringa oleifera
2.3 Use of moringa oleifera
2.4 Treating dirty water with moringa stenopetala
3.1 Fiber therm
3.2 UV spectrophotometer
3.3 Silica gel plate
3.4 Developing TLC plate
3.5 Hydraulic press
3.6 Sample for XRF analysis
4.1 Dry matter and Moisture in different parts of moringa
4.2 Ash in different parts of moringa
4.3 Fibre in different parts of moringa
4.4 Fat in different parts of moringa
4.5 Carbohydrate in different parts of moringa
4.6 Protein in different parts of moringa
4.7 Soluble sugar in different parts of moringa
4.8 Reducing Sugar in different parts of moringa
4.9 Phenol in different parts of moringa
4.10 Amino Acid in different parts of moringa
4.11 Ascorbic acid in different parts of moringa
4.12 Phytochemicals test
4.13 TLC plate of amino acid
4.14 Photograph of Antibacterial activity of different extract of moringa oleifera against salmonella typhii bacteria
4.15 Photograph of Antibacterial activity of different extract of moringa oleifera against Escherichia coli bacteria
4.16 X-Ray Florescence graph of Flower
4.17 X-Ray Florescence graph of Leaves
4.18 X-Ray Florescence graph of Seed
4.19 X-Ray Florescence graph of pulp
4.20 Heavy metal content in flower
4.21 Heavy metal content in seed
4.22 Heavy metal content in pulp
LIST OF TABLE
4.1 Carbohydrate (mg/gm) or (%)present in different parts of moringa
4.2 Protein (mg/gm) or (%) present in different parts of moringa
4.3 Total soluble sugar (mg/gm) or (%) present in different parts of moringa
4.4 Reducing sugar (mg/gm) or (%) present in different parts of moringa
4.5 Total phenols (mg/gm) or (%) present in different parts of moringa
4.6 Free Amino acid (mg/gm) or (%) present in different parts of moringa
4.7 Ascorbic acid (Vitamin C) (mg/gm) or (%) present in different parts of moringa
4.8 Result of phytochemical analysis
4.9 Result of qualitative analysis of amino acid in different parts of moringa
4.10 Zones of inhibition (mm) of salmonella typhii by different extracts of moringa parts
4.11 Sensitivity pattern of salmonella typhii bacteria for different extract of moringa parts
4.12 Element present in dry powder of moringa flower
4.13 Element present in dry powder of moringa leaves
4.14 Element present in dry powder of moringa seed
4.15 Element present in dry powder of moringa pulp
4.16 Sodium content (%) in different parts of moringa
4.17 Zinc, lead & cadmium content (mg/lit) in different sample of moringa
4.18 Permissible limits of some metals in dietary
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Moringa oleifera, an important medicinal plant is one of the most widely cultivated species of the family Moringaceae. It is highly valued from time immemorial because of its vast medicinal properties. The present study provides all necessary information regarding of four parts such as flower, leaves, seed and pulp of moringa like biochemical, phytochemical, mineral, antibacterial activity and its nutritional value. The benefits of essential nutrients and minerals for maintaining good health were also highlighted in this study.
The results of proximate analysis of Moringa oleifera revealed that the protein (9.37%), carbohydrate (7.33%), ascorbic acid (2.10%) and total soluble sugar (0.73%) were highest in flower as compared to leaves, seed and pulp. While free amino acid (9.84%) was found to be higher in seed, total phenol (0.29%) was higher in leaves and reducing sugar (0.43%) higher in pulp of the moringa . The result of qualitative analysis of amino acid represented that lysine, glycine, threonine, valine, Isoleucine, tryptophan, alanine and cystein were present in moringa.
The flower also contained higher amounts of crude fibre (0.23%) as well as moisture (90.56%), while fat (15.53%) content was found higher in seed. The dry matter (30.40%) and total ash (2.12%) content were higher in leaves. The ash content represented minerals in different amounts. The higher amount of potassium was found in flower (50.9%), seed (40.7%) and pulp (77.00%). Leaves contained higher amount of Calcium (57.18%). However Aluminum (10.00%) and Magnesium (6.07%) were found only in leaves. The result of heavy metal (zinc, lead and cadmium) and analysis represented that flower, leaves, seed and pulp have zinc (Zn), lead (Pb) and cadmium (Cd) found in lower amount then permissible limit for human body.
The results of phytochemical analysis showed that terpenoids and steroids were present in all parts of moringa. Alkaloids present only in seed. Flavonoid was present in flower and seed, saponins was present in leaves, and tannin was present in leaves and seed. Glycoside was not present in any parts (flower, leaves. seed and pulp) of moringa.
The result of antibacterial activity of different types of sample (flower, leaves. seed and pulp) of moringa showed that salmonella typhii was effectively inhibited to all the extracts studied. But Escherichia coli were not inhibited by any extract. Methanolic extract of flower, leaves, seed and pulp were highly sensitive against the salmonella typhii bacteria . Ethyl acetate and aqueous extract was less sensitive as compared to methanolic extract. So every part of moringa is said to have beneficial properties that can serve humanity. So the whole plant can be extensively studied for further research aspect.
“Moringa oleifera” is one of the 14 species of family Moringaceae, native to India, Africa, Arabia, South Asia, South America and the pacific and Caribbean Islands. Because Moringa oleifera has been naturalized in many tropic and subtropics regions worldwide, the plant is referred to number of names such as horseradish tree, drumstick tree, Ben oil tree, miracle tree, and “Mothers best friend” (Coppin, 2008).
Moringa oleifera is commonly known as “Drumstick”. It is a small or medium sized tree, about 10m height, found in the sub-Himalayan tract (Rastogi et al., 2009). Moringa oleifera is a small, fast-growing evergreen tree that usually grows up to 10 to 12m in its height, open crown of drooping fragile branches, feathery foliage of trip innate leaves and thick corky, whitish bark (Roloff, 2009).It is used as a highly nutritive vegetable in many countries. Its young leaves, flowers, seeds and tender pods are commonly consumed and they are having some medicinal properties.
Moringa oleifera is grown mainly in semi-arid, tropical and subtropical areas, corresponding in the United States. It grows best in dry sandy soil, tolerates poor soil, including coastal areas. It is a fast-growing, drought.
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Optimum leaf and pod production requires high average daily temperatures of 25-30°C (77-86°F), well-distributed annual rainfall of 1000- 2000 mm (40-80 inch), high solar radiation and well-drained soils. Growth slows significantly under temperatures below 20°C (68°F). Ideal elevation is less than 600 m (1, 970 F). Moringa oleifera is relatively tolerant of drought and poor soils and responds well to irrigation and fertilization.
The tree is wild in the Sub-Himalayan tracts from Chenab to Oudh. It grows at elevations from sea level to 1400 m. It is very commonly cultivated near houses in Assam, Bengal and peninsular India. It is a prolific coppice (Gupta, 2010). It is also cultivated in north-eastern Pakistan, north-eastern Bangladesh, Sri Lanka, West Asia and the Arabian Peninsula (Roloff, 2009).
Moringa oleifera is a small, fast-growing evergreen tree that usually grows as high as 9 m, with a soft and white wood and corky and gummy bark. Roots have the taste of horseradish. Leaves are longitudinally cracked leaves, 30-75 cm long main axis and its branch jointed, glandular at joints, leaflets are glabrous and entire. The leaflets are finely hairy, green and almost hairless on the upper surface, paler and hairless beneath, with red-tinged mid-veins, with entire margins, and are rounded or blunt-pointed at the apex and short-pointed at the base. The twigs are finely hairy and green. Flowers are white, scented in large auxiliary down panicles, pods are pendulous, ribbed and seeds are 3- angled.
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Moringa oleifera is highly valued from time immemorial because of its vast Medicinal properties. The present article provides all necessary information regarding its Phytochemical investigations, pharmacological actions and medicinal properties like anemia, anxiety, asthma, blackheads, blood impurities, bronchitis, catarrh, chest congestion, cholera, conjunctivitis, cough, diarrhea, eye and ear infections, fever, abnormal blood pressure, pain in joints, scurvy, semen deficiency, headaches and tuberculosis.
It gives an account of all the data and reports which have been appeared to prove its medicinal and nutritional importance. Its utility as a nonfood product has also been extensively described. Every part of Moringa oleifera is said to have beneficial properties that can serve humanity.
In India and other countries of the world, Phytomedicines have been used since time immemorial to treat various ailments long before the introduction of modern medicine. Herbal medicines are still widely used in many parts of the world especially in areas where people do not have access to modern medicines (Hoareau et al., 2005).
Moreover in most Asian countries where herbal medicines are still heavily relied upon because of high cost of chemotherapeutic drugs, there is a need for scientific research to determine the biological activities of medicinal plants. The findings obtained from such research may lead to the validation of traditionally used medicinally important plants and enable full usage of the properties of these plants (Adde-Mensah, 1992).
The plant is highly valued since almost every part of the plant (leaves, roots, barks, fruits, flowers etc.) is used as food with high nutritional value (Gilan et al., 2007). In addition the plant has been reported to possess antibacterial properties and this explains the reasons for its wide use in the treatment of human diseases.
Moringa oleifera is used as drug by many Ayurveda practitioners for the treatment of asthma and the Antihelminthic activity of the Methanol extract of Moringa oleifera was also noted (Singh, 2010) .
Various parts of the plants such as flower, leaves, roots, seeds, barks, fruits, pulp and immature pods act as cardiac and circulatory stimulants, possess antitumerous, antipyretic, antiepileptic, anti-inflammatory and antiulcer. It also possess carbohydrate, protein, vitamins, different amino acid, phenol and many phytochemical like saponins, tannin, alkaloids, flavonoids, glycoside, steroids.
Other important properties of the plant include diuretic (J.E.1991) , antihypertensive (M.U 1988 ), cholesterol lowering (O.D. 2003), antioxidant, anti diabetic, hepatoprotective (Rickman 1998), antibacterial and antifungal activity (M.E. 2003). India's ancient tradition of Ayurveda says the Moringa oleifera prevent 300 diseases. Our aim to identify the nutrient composition and antibacterial activity of different parts of Moringa oleifera because Moringa oleifera plant that will not only provide nutrient to the body but also help in the prevention of the diseases such as cancer, cardiovascular diseases and malnutrition.
The present investigation on “Neutraceutical, Phytochemical characterization and Antibacterial activity of Medicinal plant Moringa Oleifera ’’ was planned with following objective.
1. To estimation of biochemical parameters of different parts (flower, leaves, pulp and seed) of Moringa oleifera.
2. To determine the phytochemical of different parts (flower, leaves, pulp and seed) of Moringa oleifera.
3. To record quantative analysis of amino acids by TLC.
4. To assay antibacterial activities of different extract of Moringa oleifera.
5. To find out mineral composition of different parts of Moringa oleifera using XRF.
6. To study heavy metals in different parts of Moringa oleifera using VA.
CHAPTER-II REVIEW AND LITRATURE
2.1 HISTORY OF MORINGA OLEIFERA
The plant Moringa Oleifera was first time described in Northern India, where it was described around 2000 B.C. as a medicinal herb. The oral tradition of Ayurvedic medicine in India declared that moringa prevents more than 300 diseases.
Moringa oil was also found in ancient Egyptian culture as protection for their skin from the ravages of desert weather. Later, the Greeks found many healthful uses for moringa and introduced it to the Romans. From this time it was well known. A plantation was starting.
This well traveled plant has been carried to all the tropical parts of the world, where it readily takes root. It is commonly used for food, for medicinal purposes and clarifying water. It spread eastward from India to the lower parts of China, Southeast Asia and the Philippines. From India it also spread westward to Egypt, Africa, around the Mediterranean, and finally to West Indies in America.
Moringa has also been receiving increasing attention from food processing industries. According to the Global Facilitation Unit, 2008, African companies manufacturing cereals were interested in adding moringa leaf powder to enrich their products with a low cost, local source of vitamins and minerals.
Moringa has the potential to save millions of lives in poor countries where malnutrition is prevalent because of its high nutritional value. According to Moringa SA (2011), 80% of the production and processing costs is in manpower. This means that production of moringa for leaves and for other uses has the potential to create more job opportunities
2.2 MORINGA SPECIES
There are 14 known species of the moringa family belonging to the genus Moringaceae in the world, but only two species are distributed widely in Africa. The species are Moringa oleifera as shown in Figure 2.1 and Moringa stenopetala as shown in Figure 2.2.
The choice of moringa species depends on the intended end use. There is evidence that Moringa Stenopetala is more effective as a water purifier than Moringa oleifera (Jahn, 1988) .
However studied showed that the purification quality of Moringa oleifera changes with the 2season (Foidl et al., 2001 ). In African countries, Moringa oleifera is grown for manufacturing leaf powder while Moringa stenopetala is used for food crops.
The other twelve varieties of moringa include
1. Moringa Arborea
2. Moringa Borziana
3. Moringa Concanensis
4. Moringa Drouhardii
5. Moringa Hildebrandtii
6. Moringa Longituba
7. Moringa Ovalifolia
8. Moringa Peregrin
9. Moringa Pygmaea
10. Moringa Rivae
11. Moringa Ruspoliana
12. Moringa Stenopetala
Fig.2.1 Moringa oleifera
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Fig 2.2 Trunk and Leaves of Moringa Stenopetala
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2.3 CLASSIFICATION MORINGA OLEIFERA
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2.4 COMMAN NAME OF MORINGA OLEIFERA
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2.5 MORPHOLOGY AND PHYSICAL CHARACTERISTICS
Moringa is a slender softwood tree that branches freely and can be extremely fast growing. Although it can reach heights in excess of 10 m (33 F) and a diameter of 20-40 cm at chest height, it is generally considered a small to medium size tree.
2.5.1 STEM: The stem is normally straight but occasionally is poorly formed. The tree grows with a short, straight stem that reaches a height of 1.5-2 m before it begins branching but can reach up to 3.0 m.
2.5.2 BRANCH: The extended branches grow in a disorganized manner and the canopy is umbrella shaped.
2.5.3 LEAVES: Tripinnate compound leaves are feathery with green to dark green elliptical leaflets 1-2 cm (0.4-0.8 inch) long. The tree is often mistaken for a legume because of its leaves. The alternate, twice or thrice pinnate leaves grow mostly at the branch tips. They are 20-70 cm long, grayish-brawny when young, long petiole with 8-10 pairs of pinnae each bearing two pairs of opposite, elliptic or obovate leaflets and one at the apperoximetlly, 1-2 cm long.
2.5.4 FLOWERS: Conspicuous, lightly fragrant flowers are borne on inflorescences 10-25 cm (4-10 inch) long, and are generally white to cream colored, 2.5 cm in diameter, borne in sprays, with 5 at the top of the flower, although they can be tinged with pink in some varieties. The flowers, which are pleasantly fragrant and 2.5 cm wide are produced profusely in auxiliary, drooping panicles 10-25 cm long. They are white or cream colored and yellowdotted at the base. The five-deflexed sepals are linear-lance late. The five petals are slender-speculate. They surround the five stamens and five staminodes and are reflexed except for the lowest.
2.5.5 FRUITS: The fruits are trilobed capsules, and are frequently referred to as pods. Immature pods are green and in some varieties have some reddish color. Pods are pendulous, brown, triangular, splitting lengthwise into 3 parts when dry, 30-120 cm long, 1.8 cm wide, containing about 20 second embedded in the pith, pod tapering at both ends, 9-ribbed.
2.5.6 SEEDS: The seeds are round with a brownish semi-permeable seed hull, with 3 papery wings. Seed hulls are generally brown to black, but can be white if kernels are of low viability. Viable seeds germinate within 2 weeks. The hull itself has three white wings that run from top to bottom at 120°C intervals.
Each tree can produce between 15,000 and 25,000 seeds/year. The average weight per seed is 0.3 g and the kernel to hull ratio is 75:25.
Moringa is native to the Himalayan foothills (India/Bangladesh). As a commercial crop, it is cultivated extensively in India and parts of Africa. It would be challenging to find a region in the tropics or subtropics where moringa is not grown as a backyard tree for leaf and pod consumption, medicinally and for fiber. Moringa is most commonly found in areas with South and Southeast Asian (particularly Filipino) populations. Today it is widely cultivated in Africa, Central and South America, Sri Lanka, India, Mexico, Malaysia, Indonesia and the Philippines. It is considered one of the world’s most useful trees, as almost every part of the moringa tree can be used for food or has some other beneficial property.
The plant is propagated by planting limb cuttings 1-2 m long, from June to August. The plant starts bearing pods 6-8 months after planting, but regular bearing commences after the second year. The tree bears for several years. It does not tolerate freeze or frost. It can also be propagated by seed. As with all plants, optimum cultivation depends on producing the right environment for the plant to thrive.
Moringa is a sun and heat loving plant. Seeds are planted an inch below the surface and can be germinated year-round in well-draining soil. India is the largest producer of moringa, with an annual production of 1.1 to 1.3 million tones of tender fruits from an area of 380 km2. Among the states, Andhra Pradesh leads in both area and production (156.65 km2 ) followed by Karnataka (102.8 km2 ) and Tamil Nadu (74.08 km2 ).
In other states, it occupies an area of 46.13 km2. Tamil Nadu is the pioneering state in so much as it has varied genotypes from diversified geographical areas and introductions from Sri Lanka. Moringa is common in India, where its triangular, ribbed pods with winged seeds are used as a vegetable crop. It is particularly suitable for dry regions. The drumstick can be grown using rainwater without expensive irrigation techniques. The yield is good even if the water supply is not. The tree can be grown even on land covered with 10-90 cm of mud (Rajangam et al., 2001).
2.8 CHEMICAL COMPOSITION
The main constituents of moringa plant are palmitic and stearic acid saponins, glycoside and gum, protein Vitamins A (8855 IU per 100g), B1, B2, B3, C Minerals calcium, iron, phosphorus, magnesium. The leaves, flowers and pods are used as significant sources of vitamins A, B and C, riboflavin, nicotinic acid, folic acid, pyridoxine, ascorbic acid, beta-carotene, calcium, iron, and alpha-tocopherol (Dahot, 1988). The pods are considered good sources of the essential amino acids. A compound found in the flowers and roots of the moringa tree, pterygospermin, has powerful antibiotic and fungicidal effects (Das et al., 1957). More recently the ben oil has also been shown to be particularly effective in the manufacture of soap producing a stable lather with high washing efficiency suitable for some African countries. The root bark contains two alkaloids moringine and moringinine.
2.9 BIOCHEMICAL PARAMETER
Crude fiber of Moringa oleifera is about 4.7±0.2% (Nzikou et al., 2009) have found in their study a crude fibre value of 3.2±0.80% while (Anwar et al., 2006) have found a range value of 6.60 to 9% and (Anwar et al., 2003) a value of 7.20%. Although crude fibre does not contribute to nutrients or energy, it is a source of dietary fiber. This value of fibre might be helpful in terms of maintaining positive effects on intestine and colon physiology, besides other homeostatic and therapeutic functions in human nutrition (McPherson, 1982). Carbohydrate is the lowest macronutrient present into seeds of Moringa oleifera . Its value is about 9.17% while (Nzikou et al., 2009) have found a value of 13.6% and (Abdulkarim et al., 2005) reported a range Value of 16.5 to 17.8%. These authors have not appreciate the main carbohydrates present into seeds so, it’s difficult to compare our results to those of others authors.
The crude protein analysis by Nzikou et al., (2009) in Congo Brazzaville by the method of micro-Kjeldahl revealed a value of 37.6±1.07%. Thus, these seeds are a potentially good source of proteins and oil which should be exploited to determine if they are commercially viable.
According to several authors (Huang et al., 199 1) polyphenolics have attracted special attention because they can protect the human body from the oxidative stress which may cause many diseases.
(Odukoya et al., 2005) has reported a strong relationship between Phenolics content and antioxidant activity in selected fruits and vegetables. Thus, the presence Phenolics compounds in Moringa seed oil is an added value to its nutritional and health potential.
Phenolics compounds have the ability to scavenge free radicals and to contribute significantly towards antioxidant activities of vegetables and fruits. Plant Phenolics 6 comprise a large variety of substituted Phenolics compounds that give color and astringent taste to foods and According to (Fennema 1996) the glycosides flavonoids are the largest groups of compounds among the plant Phenolics. Phenolics acids such as caffeic, ferulic and Gallic acids and their derivatives are abundant in many plants and they include anthocyanin pigments, yellow flavones, flavonol, chalcones and isoflavones. Phenolics compounds, including their subcategory, flavonoids are present in all plants and have been studied extensively in cereals, legumes, nuts, olive oil, vegetables, fruits, tea, and red wine.
Isoprenoids also known as terpenoids are the major family of natural compounds, comprising of 40,000 different molecules. The Isoprenoids biosynthetic pathway produces both primary and secondary metabolites that are of great significance to plant growth and persistence. Terpenoids are welldefined as secondary metabolites using molecular structures comprising carbon backbones are made up of isoprene (2-methylbuta- 1, 3-diene) units. Isoprene comprises five carbon atoms and as a consequence, the number of carbon atoms in any terpenoids is a multiple of five.
The terpenoids comprises of two isoprene units, containing ten carbon atoms. Among the primary metabolites produced by this pathway are the phytohormones - abscisic acid, gibberellic acid and cytokine’s, the carotenoids, plastoquinones and chlorophylls involved in photosynthesis, the ubiquinones required for respiration, and the sterols that impact membrane structure.
Many of the terpenoids are commercially interesting because of their use as flavours and fragrances in foods and cosmetics examples menthol, nootkatone because they are important for the quality of agricultural products, such as the flavour of fruits and the fragrance of flowers like linalool (Aharoni et al., 2004) . In addition, terpenoids can have medicinal properties such as anticarcinogenic (e.g. taxol and perilla alcohol), antimalarial (e.g. artemisinin), anti-ulcer, hepaticidal, antimicrobial or diuretic (e.g. glycyrrhizin) activity (Bertea et al., 2005) . The terpenoids have also been shown to be of great ecological significance ( Degenhardt et al., 2003).
The steroids and sterols in animals are biologically produced from precursors of terpenoids. Sometimes terpenoids are added to proteins to increase their attachment to the cell membrane the process known as isoprenylation (Sacchettini et al., 1997). These compounds and their derivatives also belong to other drugs such as validol, menovasin, turpentine, bromkamfora etc. Turpentine is extensively used as external drugs and it is the main raw material for other products on the base of terpenoids.
Flavonoids are polyphenolic compounds that are ubiquitous in nature and are categorized, according to their chemical structure into flavones, anthocyanidins, isoflavones, catechism, flavonol, chalcones and flavanones.
More than 4,000 flavonoids have been recognized, many of which occur in vegetables, fruits and beverages like tea, coffee and fruit drinks.
The flavonoids have provoked considerable interest recently because of their potential valuable effects on human health they have been testified to have been shown to have several biological properties including anti- inflammatory, hepatoprotective anti-thrombotic and antiviral activities many of which may be associated, partially at least, to their antioxidant and free radical scavenging ability (Robak et. al., 1988). The antiradical property of flavonoids is directed mostly toward peroxyl and alkoxyl radicals (Husain et al., 1987). Furthermore, as these compounds present a strong affinity for iron ions their antiperoxidative activity could also be ascribed to a concomitant capability of chelating iron. (Morel et al., 1993).
One of the undeniable functions of flavonoids and related polyphones is their role in defending plants against microbial attack. This not only comprises their presence in plants as constitutive mediators but also their accumulation as phytoalexins in response to microbial attack (Grayer et al., 1994). Because of their extensive ability to prevent spore germination of plant pathogens, they have been suggested also for use against fungal pathogens. There is an ever growing interest in plant flavonoids for treating human diseases and particularly for monitoring the immune deficiency virus which is the contributing agent of AIDS.
The majority of flavonoids documented as constitutive antifungal agents in plants are flavanones, isofavonoids. The recognition that a flavones glycoside, namely luteolin 7 (200-sulphatoglucoside), is an antifungal component of the marine angiosperm Thalassic testudinum is remarkable (Jensen et al., 1998).
(Skaltsa et al., 1994) claim that acylated flavones glycosides existing in the leaf hairs of quercusilex afford the plant useful protection against the damage of UV radiation. The main experiment here was to calculate the photosynthetic proficiency of de-haired leaves.
Das 1990 have shown that a carbonyl group at C-4 and a double bond between C-2 and C-3 are also important features for high antioxidant activity in flavonoids. Butane and other 3, 4-dihydroxychalcones are more active than analogous flavones because of their ability to achieve greater electron delocalization (Dziedzicet et al., 1983). Likewise, isoflavones are frequently more active than flavones because of the stabilizing effects of the 4-carbonyl and 5-hydroxyl in the former (Dziedzicet 1983). In the antioxidant action of o- Dihydroxyfavonoids metal chelation is an important factor (Shahidi et al., 1991).
Flavonoids have been stated to possess many useful properties, containing anti-inflammatory activity, enzyme inhibition, antimicrobial activity, oestrogenic activity (Harborne et al., 1999), anti-allergic activity, antioxidant activity (Middleton et al., 1986), vascular activity and cytotoxic antitumor activity (Harborne et. al., 1992).
For a group of compounds of relatively homogeneous structure, the flavonoids inhibit a mystifying number and variety of eukaryotic enzymes and have an extremely wide range of activities. In the event of enzyme inhibition, this has been assumed to be due to the interaction of enzymes with different parts of the flavonoids molecule such as carbohydrate, phenyl ring, phenol and benzopyrone ring (Havsteen et al., 1983) .
Saponin are a group of secondary metabolites found widely distributed in the plant kingdom as plant glycosides, characterized by a skeleton resulting of the 30-carbon precursor oxidosqualene to which glycosyl residues are attached along with it they have sturdy foaming property. Conventionally, they are subdivided into triterpenoid and steroid glycosides, or into triterpenoid which are found primarily in dicotyledonous plants but also in some monocots, spirostanol, and furostanol saponins. (Hostettmann et al., 1995).
Steroid saponins occur chiefly in monocotyledons family such as the lilliaceae, agavaceae and droscoraceae and in certain dicotyledons, such as foxglove (Hostettmann et al., 1995). Potatoes are unusual because they contain both triterpenoid and steroid saponins (Price et al., 1987) .
Steroidal glycoalkaloids are found principally in members of the family belonging to Solanaceae, which includes potato and tomato. The saponins formed by tomato have been studied in detail in relation to their potential role in the defences of plants against phyto-pathogenic fungi (Osbourn, 1996).
Saponin containing plants are used as traditional medicines, especially in Asia, and are intensively used in food, veterinary and medical industries (Hostettmann et al., 1995). The pesticide activity of saponins has long been reported (Irvine, 1961) saponins-glycosides are very lethal to cold-blooded organisms, but deceptively not to mammals (Hostettmann et al., 1995). Plant extracts containing a high percentage of saponins are commonly used in Africa to treat water supplies and wells contaminated with disease vectors; after treatment, the water is safe for human drinking (Hall et. al., 1991).