PHYTOCHEMICAL SCREENING OF COCONUT WATER EXTRACT

ABSTRACT

Medicinal plants are the local heritage with global importance plants and plant based medicaments are the basis of many of the modern pharmaceuticals we use today for our various ailments. The research investigated the photochemical screening of coconut water. Coconut water was obtained fresh from coconut fruit and photochemical analysis was carried out according to standard procedures. The preliminary phytochemical analysis showed the presence of alkaloids, flavonoids, phenolics, steroids and tannins in cocos nucifera water, alycosides, phemolics, terpenoids and alkaloids were detected while others were absent. This result indicated that coconut water has useful medicinal values especially with the presence of phemolics and alkaloids.

  

TABLE OF CONTENT

CHAPTER

1.     Introduction

2.     Cocos nucifera

3.     Traditional uses of coconut

4.     Medicinal uses of coconut

5.     Economic importance of coconut

6.     Extraction, types

7.     Bioactive constituents of plants

1.     Alkaloids

2.     Saponins

3.     Flavonoids

4.     Eugenols

5.     Glycosides

6.     Phenolics

7.     Tannins

8.     Steriods

9.     Terpenes

10.                        Aim

11.                        Specific objectives

CHAPTER TWO

2.0    Materials and Methods

2.1    Reagents

2.2    Equipment/Apparatus

2.3    Methods

2.3.1 Collection of medicinal plant

2.3.2 Extraction

2.4    Phytochemical screening of the plant extracts

CHAPTER THREE       

3.0    Results and Discussion

3.1    Result of phytochemical analysis of coconut water 

CHAPTER FOUR         

4.0    Conclusion

REFERENCES    

CHAPTER ONE

1.1 INTRODUCTION

        Natural phytochemicals derived from medicinal plants have gained significant recognition in the potential management of several human clinical conditions, including cancer “Phyto” is the Greek word for plant. There are many “familiar” of phytochemicals and they help the human body in a variety of ways. Phytochemicals may protect human from a host of disease. They are non-nutritive plant chemicals that have protective or disease preventive properties, plant produce these chemicals to protect itself but recent research demonstrates that many phytochemicals can protect humans against diseases. There are many phytochemicals in fruits and herbs and each works differently. The coconut, cocos nucifera L, has been described as the “tree of life” or tree of heaven and nature’s greatest gift to man. Each part of the coconut tree can be used to produce items of value for the community. Cocos nucifera L is a dominant type of tree belonging to the family Arecaceae (palm). The common name of cocos nucifera is coconut or coconut palm. Coconut is believed to have its origins in the Ido-Malayan region from where its spread throughout to tropics. The coconut palm is monoecious, i.e. with male and female flowers on the same inflorescence, called a spadix, that develops within a woody sheathe or spathe. At flowering, the spathe splits length wise to expose the spadix. Each spadix consists of a man axis 1-1.5m (3-3-5ft) in length with 40-60 branches or spikelets bearing the flowers linder favorable growing conditions first flowering occurs about 4-5 years after planting.

        Once a palm reaches maturity, a spadix (flower spike) is produced in every leaf axil between 12 and 15 spadices are produced throughout the year at fairly regular intervals, although drought conditions can delay the emergence of the spadix or cause it to abort, the number of female flowers per spadix varies. Since the floral primordial are initiated 12 months before the spadix emerges, the number is correlated to the growing conditions (weather, nutrition) 12 months prior to emergence from the literature survey, it is quite evident that the nflowers of cocos nucifera has potent therapentic value on the area of anti bacterial, larvicidal, antioxidant, dietary anti inflammatory, hepatoprotective and anti cancer. The present investigation aims to focus on the identification of some useful phytochemicals constituents cocos nucifera water extract.

 

 

1.2 COCOS NUCIFERA

          The coconut tree (cocos nucifera) is a member of the family Arecaceae (palm family). It is the only accepted species in the genus cocos. The term coconut can refer to the entire cocnut palm, the seed or the fruit, which botanically is a drupe, not a nut. The spelling coconut is an archaic form of the word. The term is derived from the 16^th-century Portuguese and Spanish word coco meaning “head” or “skull” from the three indentations on the coconut shell that resemble facial features.

        The coconut is known for its great versatility as seen in the many uses of its different parts and found throughout the tropics and subtropics. Coconuts are the part of the daily diets of many people coconut is different from any other fruits because they contain a large quantity of “water” and when immature they are known as tender-nuts or jelly-nuts and may be harvested for drinking. When mature, they still contain some water and can be used as seed-nuts or processed to give oil from the kernel, char coal from the hard shell and coirfrom the fibrous husk. The endosperm is initially in its nuclear phase suspended within the coconut water. As development continues, cellular layers of endosperm deposit along the walls of the coconut, becoming the edible coconut “flesh” when dried, the coconut flesh is called copra. The oil and milk derived from it are commonly used in cooking and frying, coconut oil is also widely used in cooking and cosmetic. The clear liquid coconut water within is potable. The husks and leaves can be used as materials to make a variety of products for furnishing and decorating, it also has cultural and religious significance in many societies that use it.

DESCRIPTION

          Plant Cocos nucifera is a large palm, growing up to 30m (98ft) tall, with pinnate leaves 4-6m (13-20ft) long and pinnae 60-90 long old leaves break away cleanly, leaving the trunk smooth. Coconutsm are generally classified into two general types, tall and dwarf. On very fertile land, a tall coconut palm tree can yield up to 75 fruits per year, but more often yields less than 30, mainly due to poor cultural practices. Given proper care and growing conditions coconut palms produce the first fruits in six to ten years, it takes 15-230 years to reach peak production.

        Fruits botanically, the coconut fruits is a drupe not a true nut like other fruits, it has three layers the exocarp, mesocard and endocarp. The exocarp and mesocarp make up the “husk” of the coconut. Coconuts sold in the shops of nontropical countries often have had the exocarp (outermost layer) removed. The mesocarp is composed of a fiber, called coir, which has many traditional and commercial uses. The shell has three germination pores (stoma) or “eyes” that are clearly visible on its outside surface once the husk is removed.

        A full size coconut weighs about 1.44kg (3-216). It takes around 6,000 full grown coconuts to produce a tone of corpa.

        Roots unlike some other plants, the palm tree have neither a tap root nor root hairs, but have fibrous root system.

        The coconut palm root system consists of an abundance of thin rooks that grow outward from the plant near the surface. Only a few of the root penetrate deep into the soil for stability. The type of root system is known as fibrous or adventitious and is a characteristic of grass species. Other types of large trees produce a single downward-growing tap root with a number of feeder roots growing from it.

        Coconut palms continue to produce roots from the base of the stem throughout its life. The number of roots produced depends on the age of the tree and the environment, with more than 3,600 roots possible on a tree that is 60 to 70 years old. Roots are uniformly thick from the tree trunk to the root tip.

        Inflorescence: The palm produces both the female and male flowers on the same inflorescence; thus the palm is monoecious. Other sources use the term polygamomonoecious. The female flower is much larger than the male flower. Flowering occurs continuously. Coconut palms are believed to be largely cross pollinated, although some dwarf varieties are self-pollinating.

Taxonomy

·         Cellular organisms Eukaryote-Viridiplantae

·         Streptophyta –Streptophyta-Euphyllophyta

·         Spermatophyte-Tracheophyta-liliopsida

·         Commelinids – Arecales-Arecaceae

·         Arecoideae – Cocoseae- Attaleinae-cocos-Cocos nucifera

CLASSIFICATION

Kingdom            Plantae

Division              Magnoliophyta

Class                         Liliopsida

Family               Arecaceae

Genus                Cocos

Species              C. nucifera

CULTIVATION

        V palms are grown in more than 90 countries of the world, with a total production of 62 millions tones per years. Most of the world production is in tropical Asia. Coconut trees are very hard to establish in dry climates, and cannot grow there without frequent irrigation, in drought conditions, the new leaves do not open well, and older leaves may become desiccated, fruits also tends to be shed.

Technical Requirement of Coconut Cultivation

Agro-climate requirements

        Coconut is essentially a tropic plant but has been found to grow under varying agro climatic conditions. The mean annual temperature for optimum growth and maximum yield is stated to be 27⁰c with a diurnal variation of 6⁰c to 7⁰c and relative humidity more than 60%. The coconut palm thrives well up to an altitude of 600m above MSL. The coconut palm thrives well under an evenly distributed annual rainfall ranging from 100mm to 3000mm. However, a well distributed rainfall of about 2000mm is the ideal rainfall for proper growth and higher yield.

 

Soil                        

        The coconut palm can tolerate wide range of soil conditions. But the palm does show certain growth preferences. A variety of factors such as drainage, soil depth, soil fertility and layout of the land has great influence on the growth of the palm. The major soil types that support coconut in India are laterite, alluvialred sandy loan, coastal sandy and reclaimed soils with a pH ranging from 5.2 to 8.0.

Selection of site

        Soil with a minimum depth of 12m and fairly good water holding capacity is preferred for coconut cultivation. Shallow soils with underlying hard rock, low lying areas subjected to water stagnation and clayey soils should be avoided. Proper supply of moisture either through well distributed rainfall or irrigation and sufficient drainage are essential for coconut.

Preparation of land

        Size of the pit depends on the soil type and water table. In laterite soils large pits of the size 1.2m x 1.2m x 1.2 may be dug and filled up with loose soil, powered cow dung and ash up to a depth of 60cm before planting. In loaming soils, pits of size 1m x 1m x 1m filled with top soil to height of 50cm is recommended, while filling the pits, two layers of coconut husk can be arrange at the bottom of the pit with concave surface fusing upwards for moisture conservation. After arranging each layer, BHC 10%/DP should be sprinkled on the husk to prevent termite attack. In laterite soils, common salt at 2kg pit may be applied, six months, prior, on the floor of the pit to soften the hard pans.

Spacing

        In general square system of planting with a spacing of 7.5m x 7.5m is recommended for coconut. This will accommodate 177 palms per hectare. However, spacing of 7.5 to 10m is practiced in various coconut growing regions of the country.

Harvesting

        Coconuts are harvested at varying intervals in a year. The frequency differs in different areas depending upon the yield of the trees in well maintained and high yielding gardens, bunches are produced regularly and harvesting is done once a month. Coconut becomes mature in about 12months after the opening of the spathe. It is the ripe coconut which is the source of major coconut products nut which are eleven months old give fibre of good quality and can be harvested in the tracts where green husks are required for the manufacture of coir fibre. Economic life of the coconut palm is about 60 years.

1.3 TRADITIONAL USES OF COCONUT

        The coconut is used for roofing and side screens on traditional housing and woven coconut leaves are used to carry or serve food. The timber is used for housing and canoe building. While the coconut shells have been used for centuries to carry water or store coconut oil. The fresh sap is boiled to use as a natural sweetener or mixed with water to make a lovely sweet drink. Coconut trunks are used for building small bridges and huts, they are preferred for their straightness, strength, and salt resistance. Coconut trunks are used for house construction. Coconut timber comes from the trunk, and is increasingly being used as an ecologically sound substitute for endangered hardwoods. It has applications in furniture and specialized construction, as notably demonstrated in Manila’s coconut palace. The coconut husk is used as a potting medium to produce healthy forest tree sapling, coconuts are used in the beauty industry in moisturizers to its chemical structures, is readily absorbed by the skin. The coconut shell may also be ground down and added to products for exfoliation of dead skin. Coconut is also a source of lauric acid, which can be processed in a particular way to produce sodium lauryl sulfate, a detergent used in showers gels and shampoos. Half coconut shells are used in theatre foley sound effects of a horses hoof beats. Dried half shells are used as the bodies of musical instruments, coconut meals used as livestock feed. The dried calyx is used as fuel in wood fired stoves. Coconut water is traditionally used as a growth supplement in plant tissue culture/micro propagation.

1.4 MEDICINAL USES OF COCONUT

        Coconut is used to treat a wide variety of health problems including the following Abscesses, asthma, baldness, bronchitis, bruises, burns colds, constipation, cough, dropsy, dysentery, earache, fever, flu, gingivitis, gonorrhea, irregular or painful menstruation, jaundice, kidney stones, malnutrition, nausea, rash, scabies, scurvy, skin infection, sore throat, swelling, syphilis toothache, tuberculosis, tumors, typhoid ulcers, upset stomach, weakness and wounds. Coconut water is known as a tasty beverage popular mainly in tropic Island, however far not all of us are aware of health benefits of coconut water. Generally speaking, it contains a whole amount of supplements that are needed in order to supplements that are needed in order to sustain life.

·         Potassium- it helps to lower arterial blood pressure.

·         Glucose- It acts as sugar needed for body energy.

·         Vitamin C- It shields the body for ailments like scurvy.

·         Vitamin B- It helps to replace worn out tissues and cells.

Hair care

          Coconut oil is one of the best natural nutrition for hair. It helps in healthy growth of hair providing them a shiny complexion; it is effective in reducing the protein loss for damage and undamaged hair.

Immunity

        Coconut oil is also good for the immune system, it strengthen the immune system as it contains antimicrobial lipids, lauric acid, capric acid and caprylic acid which have antifungal, antibacterial and antiviral properties. The human body converts lauric acid into monolaurin which is claimed to help in dealing with viruses and bacteria causing diseases such as herpes, influenza, cytomegalovirus, and even HIV. It helps in fighting harmful bacteria such as listeria monocytogenes and heliobacterpylori, and harmful protozoa such as giardia lamblia.

 

 

Weight loss

        Coconut oil is very useful in benefits of honey in weight loss, it contains short and medium chain fattyacide that helps in taking off excessive weight it is also easy to digest and it helps in healthy functioning of the thyroid and enzymes systems. Further, it increases the body metabolism by removing stress on pancreases, thereby burning out more energy and helping obese and overweight people reduce their weight. Hence, people living in tropic coastal areas, who eat coconut oil daily as their primary cooking oil, are normally not fat, obese or overweight.   

1.5 ECONOMIC IMPORTANCE OF COCONUT

        The coconut palm provides a substantial export income for many tropical countries, as well as food and drink for home consumption and fuel and shelter of the exported products, copra, the dried kernel, is a major source of vegetable oil and coconut oil, and shielded and dried kernel is widely used in the bakery and confectionery trades as desiccated coconut. Copra cake, left after oil extraction, is a valued animal feed, especially for dairy cattle.   

          The leading coconut producer nations are (in order) the Philippians, Indonesia, India, Sri Lanka, Thailand and Malaysia, while production from nations such as Fiji, Papua New Guinea, Solomon Islands, Samoa, Tonga and Vanuatu is relatively small, the coconut trade is a major source of export revenue for these countries, in some years, copra comprises more than 50% of Vanuatu exports income.

        The coconut industry is the highest net foreign exchange earner of agricultural exports in the Philippines, accounting for about 1.5% of GNP. It employs directly or indirectly, some 20 million people (about one-third of the population) and earns more than US$510m annually. However, the industry’s ability to meet demand and expanding may be jeopardized by:

1.     The declining share of coconut oil in the world’s oil and fat market.

2.     A proposed levy on vegetable oil imports to the European community and a campaign against coconut oil in US.

3.     More stringent aflatoxin regulations imposed in the international copra market.

4.     Erosion of the European desiccated coconut market.

5.     Lack of a market development and expansion program; and.

6.     Low incomes for coconut to respond to changing patterns in world trade in coconut products could have adverse effects on employment and revenue.

1.6 EXTRACTION

        The shade-dried flowers of cocos nucifera were extracted with various solvents. Shloroform, methanol, ethanol, hydroaclcohol )80% aqueous ethanol) and aqueous extracts of flowers of cocos nuficera were prepared in 20g/200ml. the excess solvent in the extract were removed by distillation and concentrated on water bath, the extracts were then collected in petridish and stored in desicators at room temperature. The extracts were used for the detection of phytochemical analysis.

Type

          While to the consumers it may seem that there are a few different types of coconuts, there is really only one species of coconut plant. The coconut palm, cocos nucifera, is the only type of palm tree that produces coconut. Within this species, however, there are dozens of different varieties of coconuts the different varieties are usually divided into two main types, tall and dwarf.

        Tall are the most common type of coconut palm they can cross-pollinate, which means that they share genetic materials among trees, leading to a lot of variation in the characteristics of the fruit the two main types of tall coconut are the Niu Kafa which grows mostly in the wild and not commercially and the Niu Vai, which is domesticated. In most cases, the many different types of coconut that fall between the two types are named according to where they are grown some varieties include the West Africa Tall and the Tampakan Taqll.

        Dwarf coconut mostly self-pollinate, which means that there are fewer different types. As the name indicates, they are smaller than then tall, which makes them more popular to grow in home gardens and parts. Dwarf coconut trees produce more fruit than tall size do, but the coconut are generally smaller in size like tall trees, dwarfs are usually named by their country of origin, in addition, the color of then young fruits is included as part of the name. Some varieties include the Cameroon Red, the Malayan Yellow, and the Nias Green.

        One variety of coconut, the Niu Leka Dwarf, also known as the Fiji Dwarf or Samoan Dwarf, is distinct from other dwarf varieties, this several characteristics in common with the tall.

 

1.7 BIOACTIVE CONSTITUENTS IN MEDICINAL PLANTS

        Mayer’s Test: A small quantity of the extract was treated with few drops of dilute hydrochloric acid and filtered. The filtrate was filling with Alkaloid Mayer’s reagent. Formation of cream precipitate indicated the presence of alkaloids. Wagner’s Test: To 2-3ml extract with few drops Wagner’s reagent formation of reddish brown precipitate indicates the presence of alkaloids.

1.7.2 Saponins

          Foam Test: the extract was diluted with 20ml of distill water and it was shaken with a graduated cylinder for 15minutes. Alayer of foam indicated the presence of Saponnis.

1.7.3 Flavoniods

        Naoh Tests: To 2-3ml of extract drops of sodium hydroxide solution were added in a test tube formation of intense yellow colour that became colourless on addition of few drops of dilute HCL indicated the presence of flavoniods.

1.7.4 Glycosides 

        Akyl Glycosides have been described in hetero in cyanobacteria where they likel6y take part in the protection of these cells. They were isolated and studied in nitrogen-fixing cyanobacteria such as anabaena cylindrical (Nicholas BW et al, nature 1968, 217, 767) and several forms were structurally characterized.

        Other forms with different chain length and functionally groups on the chain have been studied in other species (cyanospira rippkae, anamaeba torulosa). Their structures consist of sugar moitiess glycosidically bound to long-chain diols, triols, keto-ols and keto-diols. The aglycome moiety consisted of C26 or C28 carbon-chains with hydroxyl functionalities likely at the C-3 position.

        Alkyl glucosides found in Nostacaceae were characterized by the presence of troils and of C-3 Ketones. Ascarosides are a group of simple glycolipids which were described are in eggs and tissues of nematodes (Ascaridiodea) (Fougeuy C et al, Bull Soc Biol 1957, 39, 101). Investigations on the chemical nature of ascarosides from Parascaris equorum and Ascaris Suum (Bartley PJ et al) Nat prod 1996, 59, 921) have shown that they are formed basically by a glycosyl moiety (3,6 didesoxymannose, known as ascarylose) linked to a z-hydroxylated hydrocarbon containing  26 and 33 carbon atoms.

1.7.5 Phonolics

        Phenol Test: When 0.5ml of fecl3 (W/V) solutions was added to 2ml of test solutions, formation of an intense colour indicated the presence of phenols.

1.7.6 Tannins

          Ferric Chlodride Test: Small quantity of extract was boiled in 20ml of water in a test tube and then filtered. A few drop of 0.1% ferric chloride was added and observed for brownish green or blue-black coloration which indicate the presence of tannis.

1.7.7 Steriods

        Salkowski Test: To 2ml of extract, add 2ml chloroform and 2ml concentrated H₂S05₄ and was shaken well chloroform layer appeared red and acid layer showed greenish yellow fluorescence indicated the presence of steriols.

        Liberman-Burchard’s Test: Mix 2ml extract with chloroform acid, 1-2ml acetic anhydride and 2 drops concentrated H₂S05₄ from the side of the test tube. First red, then blue and finally green colour indicated the presence of sterols.

        Bioactive constituents which have been reported include stigmastane type saponnins (vernoniosides) steroidal saponins, sesquiterpene lactones vernomygdin, hydroxy vernolide), phenolics acids lignans, xanthones, anthraquinanes, terpenese, coumarins and peptides edoties). Nutrient composition of coconut water are vitamins, glucose, potassium minerals, electrolytes, enzymes, amino acids, cytokine, and phyto-hormones.

1.7.8 Eugenols

             

Eugenol is a phenypropene, on allyl chain-substituted guaiacol. Eugenol is a member of the phenylpropaniods class of chemical compounds, it is a colourless to pale yellow oily liquid extracted from certain essential oils especially from clove oil, nutmeg, cinnamon, basil and bay leaf. It is present in concentrations of 80-90% in clove bud oil and at 82-88% in clove leaf oil.

Modern Uses of Eugenol

        Eugenol is used in perfumeries, flavorings, essential oils and in medicine as a local antiseptic and anesthetic. Eugenol can be combine with zinc oxide to form a materials known as zinc oxide engonel which has restorative and prosthodontic applications in dentistry for example, zinc oxide eugenol is used for root canal sealing it can be used to reduce the presence of listeria monocytogenes and lactobacillus sakei in food. Thet are also used in manufacturing stabilizer and antioxidants for plastics and rubbers. Attempts have been made to develop eugenol derivatives for intravenous injection, such as propanidid and G.29.505. the latter produced unacceptable side effects around the site of injection in many patients it is one of many compound that is attractive to makes of various species of orchid bees, which apparently gather the chemical to synthesize pheromones, it is commonly used as bait to attract and collect these bees for study. It also attract male cucumber beetle it is recently discovered that eugenol and isoeugenol, flora volatile scent compound, are catalyzed by single type of enzyme in gymnadenia species and gene encoding for this enzyme is first functionally characterized gene in this species so far.

        It is commonly used in wisdom tooth extraction surgeries complicated by dry socket. Clove oil is growing in popularity as an anesthetic for use on aquarium fish as well as a wild fish when sampled for research and management purposes. Where readily available, it presents a humane method to euthanize sick and diseased fish either by direct over dose or to induce sleep before and overdose of ethanol; it is used in some mousetraps and skills certain human colon cancer cell lines in vitro and in vivo. Eugenol may have potential therapeutic effects against diseases characterized by excessive osterclast activity.

TOXICITY OF EUGENOL

          Eugenol is hepatotouix, meaning it may cause damage to the liver. Overdose is possible, causing a wide range of symptoms from blood in the patient’s urine to convulsions, diarrhea, nausea, unconsciousness, dizziness or rapid heartbeat. According to a published 1993 report, a 2 year old boy nearly died after taking between 5 and 10ml eugenol is subject to restrictions on its use in perfumery as some people may become sensitized to it, however, the degree to which eugenol can cause an allergic reaction in humans is disputed.

ALLERGY OF EUGENOL

        Eugenol is a component of Balsam of Peru to which some people are allergic. When eugenol is used in dental preparations such as surgical pastes, dental packing, and dental cement, it may cause contact stomatitis and allergic cheilitis. The allergy can be discovered via patch test.

1.7.9 Terpenes

        Terpene, any of a class of hydrocarbons occurring widely in plants and animals and empirically regarded as built up from isoprene a hydrocarbon consisting of five carbon atoms, (C₅ H₈). The term is often extended to the terpeniods, which are oxygenated derivatives of these hydrocarbons.

        Biological formation of the terpenes occurs by the combination of two molecules of acetic acid to give mevalonic acid (C₆ H₁₂ 0₄) and conversion of the latter to isopentenyl pyrophosphate, which contains the  five carbon isoprene skeleton.  Further transformation of the isopentenyl compound yield the true terpenes and the terpenoids.

        The true terpenes are usually grouped according to the number of isoprene (C5 H8) units in the molecule: monoterpenes (C10 H16) contains two such units; sesquisterpenes (C15 H24), three; diterpenes (C20 H32), four; triterpenes (C30 H48), six; and tetraterpenes (C40 H64), eight. Rubber and gutta-percha are polyterpenes in which 1,000-5,000 isoprene units are joined in a long chain. Monoterpenes, sesquiterpenes, and diterpenes are abundant in essential oils of plants: turpentine containe contains several monoterpenes. Vitamin A is another important diterpene. The triterpene  squalene obtainable from shark-liver oil, may be converted to cholesterol and many other steroids. The carotenoid pigments are the best known tetraterpenes.

1.10 AIM

        The aim of this work is to identify the phytochemicals present in cocos nucifera water extract.

1.11 Specific Objective

        The specific objectives are to carry out phytochemical analysis on coconut water and also:

1.     Collecting drying and pulverisationof coconut water.

2.     Extraction of coconut water using distilled water by maceration method.

3.     Determination of phytochemicals present in extract of the plant.