Investigatory Project

San Miguel National High School Scuala St., San Juan, San Miguel, Bulacan

Phytochemical Screening of Papua Leaves (Polyscias fruticosa) An Investigatory Project Presented To The Department Of Science – San Miguel National High School

In Partial Fulfillment Of The Requirements In Technical Writing II

Submitted to: Mrs. Marinella P. Garcia Sy

Submitted by: Princess Fay S. Lopez Mark Ranniel M. Estarez IV- Science Class (B) October 2012

ABSTRACT The importance of plants is known to us well. The plant kingdom is a treasure house of potential drugs and in the recent years there has been an increasing awareness about the importance of medicinal plants. Drugs from the plants are easily available, less expensive, safe, and efficient and rarely have side effects. Papua Plant Leaves were air dried and washed with distilled water. It was cut into pieces and turned into smaller pieces using the domestic blender. The extract was obtained and transferred to Beaker using cheese cloth. The extract was then screened for the presence of nine phytochemicals which were alkaloids, carbohydrates, glycosides, saponins, phenols, tannins, anthocyanin, protein and flavonoids. The result revealed the abundance of flavonoids, saponins and proteins. There were traced amounts of carbohydrates, alkaloids, phenols and anthocyanin. However, glycosides and tannins were absent in the plant extract. Based on the results of the phytochemical screening, properties and uses of Papua Plant extract could be used in the industry such as foaming agents, wood and plastic production as well as other chemical processes.

ACKNOWLEDGEMENT

The researchers would like to express their deepest gratitude and heartfelt appreciation to the people who showed their support throughout the completion of this study and also to the people who shared their time and their God-given talents to accomplish this study. To name a few: Mrs. Marinella P. Garcia Sy, the research adviser, who has been there in every step of the research, for her strong encouragement to pursue the study even during the most critical times; Ms. Farrah O. Macapagal for extending their never- ending support and sharing every knowledge and for helping the researcher complete the study; Ms. Amelia G. Quizon, the class adviser, who has been there and always ready to help the researchers, and for her moral support and motivation; Mrs. Lucena T. Serrana and Mrs. Luchie Muico, for assisting the researchers and for giving them the permission to use the Bio-Laboratory; Mr. Rosauro A. Villanueva and Ms. Lydia V. Cheng for their encouragement and financial aids; Ms. Celine Kate T. Balin, Ms. Vernikka Naomi A. Mangaluz, Mr. Ahrby R. Estarez, Mr. Nathan Yosef V. De Jesus, Ms. Jeramy Raizza May D. Espinar, Ms. Meryll M. Manalastas, the co-researchers who have been there since the start of the study, always ready to help and for giving the researchers the support and encouragement they need; IV- Special Science Class (B) students for bringing back the joyful and encouraging spirit every time things got difficult, for lending a helping hand every time the researchers need some. Without them, the researchers would not have been inspired with what they are doing; The Parents of the Researchers who gave their unparalleled support until the completion of the study;

And above all, to the Almighty God, the Guiding Light, who gave every reason why the researchers continue the project despite circumstances, for guiding them and making things easy to carry, and also reminding them that success comes with big challenges.

TABLE OF CONTENTS Title Page ………………………………………………………………………………………….... …... i Table of Contents ……………………………………………………….…………………………........ ii Abstract.………………………………………………………………………….……….......................iii Acknowledgement ……………………………………………………………………………...…….....iv INTRODUCTION Background of the Study …………………………………………………………………..….. 1 Objective of the Study ……………………………………………………..…………………. .2 Significance of the Study ………………………………………………………………...…….2 Scope and Limitations …………………………………………..…………………………….. 3 Definition of Terms ………………………………………………………………………..…… 4 REVIEWOF RELATED LITERTURE Papua (Polyscias fruticosa)……..………………………………...……………….……..…. 6 Phytochemicals ………………………………………………………………………………… 8 Related Studies ………………………………………………………………..………………13 METHODOLOGY Research Model ……………………………………………………………………….…..…..18 Phytochemical Profiling………………………………………..…………………..….………20 Materials ………………………………………...…………………………………..... 20 Procedure ……………………………………………………………………………...20 Preparation of Extract …………………………………………..……...…...20 Preparation of Reagents …………………………………………….….….. 21 Phytochemical Screening ………………………………………………….30

RESULTS AND DISCUSSION Phytochemical Analysis of Papua Leaves Pure Extract …………………….……….……39 SUMMARY, CONCLUSION AND RECOMMENDATIONS ………………………………....…….41 BIBLIOGRAPHY………………………………………………………….........................................43 APPENDIX …………………………………………………………...............................................45

INTRODUCTION The importance of plants is known to us well. The plant kingdom is a treasure house of potential drugs and in the recent years there has been an increasing awareness about the importance of medicinal plants. Drugs from the plants are easily available, less expensive, safe, and efficient and rarely have side effects. The plants which have been selected for medicinal use over thousands of years constitute the most obvious choice of examining the current search for therapeutically effective new drugs such as anticancer drugs, antimicrobial drugs, antihepatotoxic compounds.

Plants would be the best source to obtain variety of drugs. About 80% of individuals from developed countries use traditional medicines, which has compounds derived from medicinal plants. However, such plants should be investigated to better understand their properties, safety, and efficiency. Medicinal plants contain some organic compounds which provide definite physiological action on the human body and these bioactive substances include tannins, alkaloids, carbohydrates, terpenoids, steroids and flavonoids. These compounds are synthesized by primary or rather secondary metabolism of living organisms. Secondary metabolites are chemically and taxonomically extremely diverse compounds with obscure function. They are widely used in the human therapy, veterinary, agriculture, scientific research and countless other areas. A large number of phytochemicals belonging to several chemical classes have been shown to have inhibitory effects on all types of microorganisms in vitro. Plant products have been part of phytomedicines since time immemorial. This can be derived from barks, leaves, flowers, roots, fruits, seeds. Knowledge of the chemical constituents of plants is desirable because such information will be value for synthesis of complex chemical substances.

Statement of the Problem This study on “Phytochemical Screening of Papua (Polyscias fruticosa) Leaves” aims to accomplish the following objectives: 1. To determine the presence or absence of alkaloids, carbohydrates, glycosides, saponins, phenols, tannins, anthocyanin, flavonoids, and protein in the pure extract of Papua leaves.

2. To determine qualitatively the phytochemical constituents present in the pure extract of Papua leaves as absent, traced amount, slightly detectable and strongly present. 3. To determine possible medicinal activities based on the presence of phytochemicals as revealed by the phytochemical screening. 4. To determine possible uses of Papua leaves in industrial processes based on the abundance of phytochemicals as revealed by the phytochemical screening.

Significance of the Study Plant-derived substances have recently become of great interest owing to their versatile applications. Plants with medicinal uses have been proven to be the richest source of traditional and modern medicines and drugs. An enormous variety of medicinal plants are used worldwide by about 80% of the world population, although in most cases no scientific studies have been done to prove the efficacy of these medicinal plants. Furthermore, plants have been also playing a role in industry as its phytochemical constituents are utilized to produce products like dye, foams, and plastics. The screening of the phytochemical components of Papua leaves can be a scientific standard to discover new plants that contains important bioactive components. The study’s aim is designed to assess the profile of Neem leaves when it comes to its phytochemistry.

Phenols are known to have a part in acting as an antimicrobial, anthelmintic or antidiarrheal and are important to chemical industry, engineering, and wood and plastic processing. Flavonoids, tannins, and alkaloids are also playing a role in inhibiting growth of microbes. On the other hand, saponins and glycosides are capable of being an antidiarrheal

agent. In industry, they are essential in providing foaming agents. Anthocyanin can be used as pH indicators while protein and carbohydrates are parts of a healthy cell. The results can contribute to the development of new generalizations regarding new plant that have important phytochemical constituents. It can also be a stepping stone for more sophisticated quantitative analysis of exact amount of the phytochemicals detected in the plant. Thus, providing recommendations for further study is a part of the goal of this project.

Scope and Limitation This study was limited to the use of natural Papua leaves as a source of pure extract to be tested. The phytochemical properties of this plant were tested to determine only the presence of alkaloids, carbohydrates, glycosides, saponins, phenols, tannins, anthocyanin, protein and flavonoids. Extraction of Papua leaves was conducted at San Miguel National High School’s Bio-laboratory, SEDP building, San Miguel, Bulacan. Chemicals needed for the screening were purchased from University of the Philippines Diliman at Quezon City. Preparation of the agents, as well as the phytochemical profiling was done at San Miguel National High School’s Bio-laboratory, SEDP building, San Miguel, Bulacan. The results were only to determine qualitatively the presence of each phytochemical component as absent, with traced amount, slightly detectable or strongly present.

Definition of Terms 

Alkaloids – highly reactive substances with biological activity in low doses and contains nitrogen. It is bitter tasting, generally white solids and give a precipitate with heavy metal iodides.

 Anthocyanins – versatile and plentiful flavonoids pigments found in red or purplish fruits and vegetables, including purple cabbage, beets, blueberries, cherries, raspberries and purple grapes. Within the plant they serve as key antioxidants and pigments contributing to the coloration of flowers.  Carbohydrate –most abundant class of organic compounds found in living organisms. They originate as products of photosynthesis, an endothermic reductive condensation of carbon dioxide requiring light energy and the pigment chlorophyll.  Glycoside – any compound containing a carbohydrate molecule (sugar), particularly any such natural product in plants, convertible, by hydrolytic cleavage, into a sugar and a nonsugar component (aglycone), and named specifically for the sugar contained, as glucoside (glucose), pentoside (pentose), fructoside (fructose), etc.  Phenols – class of chemical compounds consisting of a hydroxyl group (-OH) bonded directly to an aromatic hydrocarbon group.  Phytochemicals – chemicals that are produced by plants. Currently, the term is being used only for those plant chemicals that may have health-related effects but are not considered essential nutrients.  Phytochemical Screening – process of tracing plant constituents. There is a standard procedure which is usually involving color changes as indicators.  Protein – any of a group of complex organic compounds which are the most important constituents of protoplasm and is composed of amino acids combined in a large molecule.

 Saponins – glucosides with foaming characteristics. Saponins consist of a polycyclic aglycones attached to one or more sugar side chains.  Tannin – any of a group of pale-yellow to light-brown amorphous substances in the form of powder, flakes, or a spongy mass, widely distributed in plants and used chiefly in tanning leather dyeing fabric, making ink, and in various medical applications.  Sudorific Inhalant - a sudorific agent. Sweat glands are stimulated by cholinergic drugs. The alkaloid pilocarpine is a potent sudorific drug, but it is rarely used for that purpose in modern medicine.

REVIEW OF RELATED LITERATURE I.

Papua (Polyscias fruticosa)

A. Description An erect shrub native from India to Polynesia, growing from 1 to 2.5 meters high. Leaves are compound, pinnate, and up to 30 cm long. The pinnae are 6 to 10, the upper ones are shortedd. Leaflets and ultimate segments are very diverse, mostly lanceolate, 5 to 10 cm long; the terminal segments are usually larger than the others and more often lobed, pointed at the tip, sharply and irregularly toothed. Flowers are numerous, umbellately arranged, shortly stalked, borne on terminal inflorescences in the upper axils of the leaves, up to 15 cm long. Fruit is broadly ovoid, compressed and about 4 cm long. Papua needs full sun to partial shade to high interior lighting. In the greenhouse, we use a soil mix consisting of 2 parts peat moss to 2 parts loam to 1 part sand or perlite. The plants should be kept moist and never should be allowed to dry thoroughly. Plants are fertilized only 3 times during the growing season using a balanced fertilizer diluted to ½ the strength recommended on the label. Since the plants are fairly slow growers, very little pruning is needed to keep the desired form. During the winter months, water should be restricted, but the plant should never be allowed to dry out completely.

B. Traditional medicinal use Gastrointestinal Diseases

The leaves in the form of a decoction are used to treat a variety of gastric maladies (tympanism, dyspepsia, flatulence and diarrhoea). In Malaysia it is commonly used to treat haemorrhoids where the leaves are finely powdered as a poultice and shaped into an elongated mass to be inserted per rectal at bedtime. The roots and twigs are used for cleaning gums, teeth and relieving mouth ulcers. Renal Disease The plant is belived to have diuretic properties and it is used in the treatment of kidney diseases especially stones.The part which is most active is the roots. For cleansing the kidneys Indonesian traditional practitioners recommend extracting the juice of the fresh leaves. Other Uses In China and Indo-China it is considered a sudorific and febrifuge and it used in the treatment of fever. It also has astringent and anti-inflammatory properties and for this it is used to help ally rheumatic pains and neuralgia. The leaves can be eaten raw or cooked as vegetable. It is also used to flavor meat and fish because it has the taste of parsley. Phytochemicals The study of phytochemicals that are derived from plants is known as phytochemistry. Description of various metabolic compounds can be useful for understanding plant functionality. As these secondary metabolites are known to protect plants from insects and various diseases affecting plants phytochemical studies are important. The extraction of phytochemicals and their isolation for use are common practices. The phytochemical properties of plants are helpful in treating medical conditions. They could be a part of the industry, as well. A Alkaloids

Alkaloids are found primarily in plants and are especially common in certain families of flowering plants. More than 3,000 different types of alkaloids have been identified in a total of more than 4,000 plant species. The function of alkaloids in plants is not yet understood. It has been suggested that they are simply waste products of plants’ metabolic processes, but evidence suggests that they may serve specific biological functions. In some plants, the concentration of alkaloids increases just prior to seed formation and then drops off when the seed is ripe, suggesting that alkaloids may play a role in this process. Alkaloids may also protect some plants from destruction by certain insect species. The medicinal properties of alkaloids are quite diverse. Certain alkaloids act as cardiac or respiratory stimulants. Quindine, which is obtained from plants of the genus Cinchona, is used to treat arrhythmias, or irregular rhythms of the heartbeat. Many alkaloids affect respiration, but in a complicated manner such that severe respiratory depression may follow stimulation. It also acts as antimicrobial, antihelmintic and antidiarrheal.

B Carbohydrates Carbohydrates are one of the main types of nutrients. They are the most important source of energy for your body. Your digestive system changes carbohydrates into glucose (blood sugar). Your body uses this sugar for energy for your cells, tissues and organs. It stores any extra sugar in your liver and muscles for when it is needed. Carbohydrates are called simple or complex, depending on their chemical structure. Simple carbohydrates include sugars found naturally in foods such as fruits, vegetables, milk, and milk products. They also include sugars added during food processing and refining. Complex carbohydrates include whole grain breads and cereals, starchy vegetables and legumes. Many of the complex carbohydrates are good sources of fiber. C Glycosides

Glycosides are believed to serve several purposes in the plant. Glycosides are bitter tasting, and it is believed that they help keep birds and insects from eating seeds and fruit before they are fully grown, by which time the glycosides have been converted to sweet sugars. Furthermore, glycosides are essential to industry. Among the important glycosides are indican, used for dyeing; digitalin, used in medicine; and the saponins, foaming agents used industrially and medicinally. D Saponins The name saponin is derived from the Latin word ’sapo’, which means the plant that consists of frothing agent when diluted in aqueous solution. Saponins comprise polycyclic aglycones. The sapogenin or the aglycone part is either a triterpene or steroid. The combination of sapogenin, hydrophobic or fat-soluble, hydrophilic or water-soluble sugar part enhances the foaming ability of saponins. Some toxic saponins are known as sapotoxin. Saponins are used on injection, for which it has a pharmacological reputation. It results in the lysis of the blood cells, haemolysis, like all detergents, and is therefore highly toxic. Saponins base are the basic of many arrow poisons. The best interesting part to be noted is that, saponins have always been toxic to cold-blooded creatures like snake and/or fish. Saponins have been also used as cleaning agents and as foam producers, notably in fire-extinguishing fluids. They have a bitter taste and when ingested orally are practically nonpoisonous to warm-blooded animals.

E.

Phenols In organic chemistry, phenols are a class of chemical compunds consisting of a hydroxyl

group (—OH) bonded directly to an aromatic hydrocarbon group. Phenols are found in the natural world, especially in the plant kingdom. In some cases of natural phenols, they are present in vegetative foliage to discourage herbivory. The class of phenols (or phenoles) is an

important raw material and an additive for industrial purposes in: laboratory processes, chemical industry, chemical engineering processes, wood processing, and plastics processing. E Tannins These are also tannin acid, common name applied to a group of vegetable products, amorphous and crystalline, obtained from various plants, and important commercially in the tanning of leather. Tannins have variable composition. Some, called condensed tannins, are phenols of moderately complex structure, and others are esters of glucose or some other sugar with one or more trihydroxybenzoic acids. The empirical formula, C 14H14O11, often given for common tannin, is only an average. Tannins occur in many trees, and the best sources include oak galls and the bark of sumac. Extraction with water, or water and alcohol, is the first step in the preparation of tannin. Settling, followed by evaporation at a low temperature, yields the commercial product. F Anthocyanins Anthocyanins

can

be

used

as pH

indicators because

their

color

changes

with pH.Anthocyanins are pink in acidic solutions (pH < 7), purple in neutral solutions (pH ~ 7), greenish yellow in alkaline solutions (pH > 7), and colorless in very alkaline solutions where the pigment is completely reduced. In photosynthetic tissues (such as leaves and sometimes stems), anthocyanins have been shown to act as a sunscreen, protecting cells from high-light damage by absorbing bluegreen and UV light. In addition to their role as light-attenuators, anthocyanins also act as powerful antioxidants. However, it is not clear whether anthocyanins can significantly contribute to scavenging of free radicals produced through metabolic processes in leaves, since they are located in the vacuole and, thus, spatially separated from metabolic reactive oxygen species. Some studies have shown that hydrogen peroxide produced in other organelles can be neutralized by vacuolar anthocyanin. G Proteins

Proteins are any of a group of complex organic compounds which are the most important constituents of protoplasm and are composed of amino acids combined in a large molecule. They are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. Protein is a part of every cell in the body, and no other nutrient plays as many different roles in keeping people alive and healthy. The importance of protein for the growth and repair of muscles, bones, skin, tendons, ligaments, hair, eyes and other tissues is proven since a very long time. Without it, the body would lack the enzymes and hormones needed for metabolism, digestion and other important processes. H Flavonoids Flavonoids are water soluble polyphenolic molecules containing 15 carbon atoms. Flavonoids belong to the polyphenol family. Flavanoids can be visualized as two benzene rings joined together with a short three carbon chain. One of the carbons of the short chain is always connected to a carbon of one of the benzene rings, either directly or through an oxygen bridge, thereby forming a third middle ring, which can be five or six-membered. The flavonoids consist of six major subgroups: chalcone, flavone, flavonol, flavanone, anthocyanins and isoflavonoids. Together with carotenes, flavanoids are also responsible for the coloring of fruits, vegetables and herbs. It has many health promoting effects. Some of the activities attributed to flavonoids include: anti-allergic, anti-cancer, antioxidant, anti-inflammatory and anti-viral. The flavonoids quercetin is known for its ability to relieve hay fever, eczema, sinusitis and asthma. Epidemiological studies have illustrated that heart diseases are inversely related to flavonoid intake. Studies have shown that flavonoids prevent the oxidation of low-density lipoprotein

thereby

reducing

the

risk

for

the

development

of

atherosclerosis.

The contribution of flavonoids to the total antioxidant activity of components in food can be

very

high

because

daily

intake

can

vary

between

50

to

500

mg.

Red wine contains high levels of flavonoids, mainly quercetin and rutin. The high intake of red

wine (and flavonoids) by the French might explain why they suffer less from coronary heart disease then other Europeans, although their consumption of cholesterol rich foods is higher (French paradox). Many studies have confirmed that one or two glasses of red wine daily can protect against heart disease. Tea flavonoids have many health benefits. Tea flavonoids reduce the oxidation of lowdensity lipoprotein, lowers the blood levels of cholesterol and triglycerides.Soy flavonoids (isoflavones) can also reduce blood cholesterol and can help to prevent osteoporis. Soy flavonoids are also used to ease menopausal symptoms.

II.

Related Studies

“On The Antipyretic, Anti-Inflammatory, Analgesic And Molluscicidal Properties Of Polyscias Fruticosa (L) Harms” Bensita Mary Bernard, Nilani Pakianathan, And Madhu C. Divakar The n-butanol extract of the leaves Polyscias fruticosa (L) Harms (Araliaceae) was tested for its anti-inflammatory activity plethismometrically in egg white induced paw oedema in rats, antipyretic activity and analgesic activity by writhing method phenyl butazone, paracetamol and aspirin were used as positive controls for anti-inflammatory, antipyretic and analgesic activity screening studies receptivity. It as observed that the n-butanol fraction mainly contains terpenoid type of saponins and designated as NBES fraction- (n –butanol extract containing

saponins). Molluscicidal screening studies proved the effectiveness of NBES to control certain kind of snails which are considered as the primary host of fluke worms. “A Pharmacognostic Report On The Leaf And Root Of Polyscias Fruticosa (L.) Harms” Bensita Mary Bernard, Nilani Pakianathan, R. Venkataswamy, And Madhu C. Divakar The major diagnostic characters and qualitative chemical and physical tests responsible for the pharmacognostic identity of the leaf and of root of Polyscias fruticosa (L) Harms have been reported. Literature survey showed the absence of any systematic pharmacognostic studies for this plant. Polyscias Fruticosa-Medicinal Tonic

The leaves are deeply lobed and double-serrate margins irregularly. Place the leaves are fragrant. The inflorescence is a short form blocks, including multiple scattering chùy collaborations. Each bearing numerous small flowers foliage has short-stalked. Flowers white egg-shaped, 5 petals and 5 stamens with long filaments recharge with a short and slender, elected under the two fields have white pale edges. Fruit Oblate silvery 3conductor 0.4 mm long, thick, carries hose exist. About 40-50 years, many scientists around the world have to pay attention to effects increase the force, added body of many trees along with ginseng plants. Some trees in the family for the tonic is well-known and used from people in longstanding us national team, such as ginseng, polyscias fruticosa also has additional effects such as many relatives to it.

Old days in Duvall often organized wrestling, before wrestling wrestlers or decision leaves polyscias fruticosa with water to drink for increased toughness, perennial indefatigable. Parts used to make drugs mainly roots, taking in the crops from 3 years upwards. People often dig taking root tree polyscias fruticosa in autumn or winter because at this active focus on the roots and roots softer. Root digging about bringing clean sandy soil, finely chopped and then drying, or drying. Years ago scientists of our country (National Institute of military medicine) there are also many studies use the polyscias fruticosa do drugs boost, increasing the likelihood of labour for persons with good results. Do supplements cause appetite, slept well, gaining weight, helps the body quickly recovered after surgery, ill. Polyscias fruticosa used quite safely. The average dose is 0.25-0, 50 g once, on drinking 2-3 times, taking the form of drug powder (star Jasmine, kibble, Sieve flour smooth), pill, drug or alcohol. In addition a number of studies also showed polyscias fruticosa is a sedative and increases the effect of anti-malarial. Among the people there were also used to treat coughs, polyscias roots through minor, milk and curing dysentery. Or use leaves polyscias fruticosa giã ragged to fill wounds. Song notes distinguish small leaf or tree crops polyscias fruticosa gỏi cá above with some trees similar: polyscias fruticosa leaves rounded, polyscias fruticosa bloom ... tonic effects, not appointed.

Studies on The Adaptogenic and Antibacterial Properties of Polyscias Fructicosa (L) Harms M.B. Bensita, P.Nilani, S. Sandhya M Pharmacognosy And Photochemistry Laboratory, College Of Pharmacy, Sripms, Coimbatore, Tamil Nadu – 641 04.

Received: 8th February, 1998 Accepted: 11th August, 1998 In the present study the adaptogenic activities of the saponin fractions of the leaves and roots of Polyscias fruticosa (L) Harms (Araliaceae) were studied in comparision wit white panax ginseng root saponins. The antibacterial activities of the polyacetylenic compound in leaves were also studied. The adaptogenic activity studies showed that polyscias fruticosa leaf and root saponins have effective abtistress activity as compared with the white Panax ginseng root saponins. The antibacterial study revealed that the polyacetylenic fraction present in polyscias fruticosa leaf and root saponins have effective abtistress activity as compared with the white panax ginseng root saponins. The antibacterial study revealed that the polyacetylenic fraction present in polyscias fruticosa leaves has got better antibacterial property compared to the saponin fraction. Mainly adaptogenic drugs are used to enhance immunity against diseases, reduce mental stress and strain, impart a euphoric effect, retard ageing processes etc., sometimes these drugs are used as a tonic to gain non-specific resistance against various ailments. Members of the araliaceae family contain triterpenoid saponin. Many chemical investigations on the triterpenoid saponins of the members of the members of the family revealed that the saponin content in these plants play an important role in pharmacological activities like stimulation of CNS. Reduction of fatigue ad enhancement of non-specific resistance. Literature survey showed that five polyactylenic alcohols are present in the root of ployscias frusticosa7,8. We have isolated these polyacetylenic fraction and screened its antimicrobial activity. The n-butanol fraction (mainly containing triterpenoid type of saponins) was screened for adaptogenic activity by set oexperiments like forced locometer activity, beavioural despair test, righting reflux test, swimming performance test, hypoxia test, hypethermia test immobilization stress ulceration, anabolic effect and immunostimulant activity. These activities of polyscias fruticosa saponins were compared with those of the root saponins of white panax gingseng.

DIURETIC ACTIVITY OF Polyscias Fruticosa (L.) HARMS *R. Varadharajan and D. Rajalingam *Kamalakshi Pandurangan College of Pharmacy, Ayyampalayam, Tiruvannamalai, Tamil Nadu 606 603.

Polyscias fruticosa (L.) Harm (Family: Araliaceae) is possibly native to Malaysia, but now widely cultivated in tropical areas and as a greenhouse plant. In Fiji, the root is used as a diuretic. The juice from the bark is taken for thrush and an ulcerated tongue or throat. A poultice made from the bark is used on syphilitic sores. The present study was undertaken to investigate diuretic effect of petroleum ether extract of the Polyscias fruticosa (L.) Harm (PEPF) in albino rats. Acute oral toxicity study was performed as per OECD guidelines. In acute oral toxicity study, mortality was not observed up to 2000 mg/kg bodyweight. PEPF were administered at the doses of 250 and 500 mg/kg, p.o. Furosemide (500 mg/kg, p.o) was used as positive control in study. The diuretic effect of the extract was evaluated by measuring urine volume, sodium and potassium content. Urine volume is significantly increased at two doses of PEPF 250 & 500 mg/kg body wt in treated rats. The excretion of sodium, Potassium levels was also increased by the PEPF. The diuretic effect of the extract was similar to furosemide. The PEPF had the additional advantage of chloride conserving effect. This study concludes that PEPF produced notable diuretic effect which appeared to be comparable to that produced by the standard diuretic furosemide. The present study provides a quantitative basis for explaining the folkloric use of Polyscias fruticosa (L.) Harm as a diuretic agent. The Leaf Volatile Oil of

Polyscias Fruticosa (L.) Harm

Oliveros-Belardo, Luz et al

The study was conducted to identify the components of the volatile oil of the leaves of N. frticosum, particularly sesquiterpenes for which the oil gave a remarkably positive test. Hydrosteam distillation of the fresh leaves of N.fruticosum gave a 0.32% yield of volatile oil. The oil was light yellow in color with a grassy note scent. Its refractive index was 1.5001 at 25oC.Tests revealed that the oil is negative for the presence of sulfur and nitrogen but highly positive for the presence of sesquiterpene. Gas liquid chromatography coupled with mass spectroscopy analysis of the oil showed the presence of ∞-bergamotene, oxygenated sesquiterpene C15H16O2 1-ethyl-1-methyl-1-2-(1-methyl1-ethenyl)-4-(1-methylethylidene)cyclohexane, ∝-elemene, β-bourbonene, β-cubebene, β-bisabolene, ∝-farnesene, ∝-elemene, β-elemene, ∝-cadinene, ∝-elemene, ∝-copaene. Further toxicity tests were recommended in order to determine its feasibility as a legitimate spray pesticide for house insect pests

METHODOLOGY

Block Diagram

Gathering of Leaf Source (Papua Tree)

Air Drying of Papua Leaves

      

Mayer’s Reagent Molisch’s Reagent Acetic Acid Ferric Chloride Distilled Water Ferric Chloride Solution Sodium Hydroxide



Solution Nitric Acid

Preparation of Pure Extract



Washing the Leaves

 

with Distilled Water Cutting of Leaves Pounding the leaves



using mortar and pestle Extraction of Leaves



using Cheese Cloth Filtration of Extract



using Filter Paper Preparation of Reagents for Phytochemical Profiling

I. Phytochemical Profiling A. Materials Papua Plant Leaves Distilled Water Scissors Domestic Blender Cheese Cloth Beaker Medicine Dropper B. Procedure

Test Tubes Graduated Cylinder Petri dishes Mercuric Chloride Potassium Iodide Naphthol Ethanol

Acetic Acid Ferric Chloride Sodium Hydroxide Nitric Acid Hydrochloric Acid

B.1. Preparation of Pure Extract, Reagents and Solutions Papua Plant Leaves were air dried and washed with distilled water. It was cut into pieces and turned into smaller pieces using the domestic blender. The extract was obtained and transferred to Beaker using cheese cloth. q

Fig 6 Pure Papua Plant Extract B.2. Preparation of Reagents, Solutions and Chemicals for Phytochemical Profiling B.2.1 Prepare the Chemicals Needed

Fig. 7 Chemicals Needed B.2.2 Mayer’s Reagent (Reagent for Alkaloid Test) 0.4 g of Mercuric Chloride (HgCl2) is weighed and dissolved in 15 mL of water (H2O). On one hand, 1.5 g of Potassium Iodide (KI) is dissolved in 2.5 mL of water (H 2O). Sufficient water is added to make the volume 25 mL A fraction of the reagent was obtained and placed in a test tube.

Fig. 8 0.4 g Mercuric Chloride

Fig. 9. Mercuric Chloride Aqueous Solution

Fig. 10 1.5 g Potassium Iodide

Fig. 11. Potassium Iodide Aqueous Solution

Fig. 12 Mayer’s Reagent B.2.3 Molisch’s Reagent (Reagent for Carbohydrate Test) A mass of 3.8 g of naphthol was obtained and dissolved in 25 mL of ethanol. A fraction of the reagent was transferred into a test tube.

Fig. 13 3.8 g of Naphthol

Fig. 14 Naphthol to Water Fig. 15. Molisch’s Reagent B.2.3 Acetic Acid and Ferric Chloride (Chemicals for Glycoside Test) 2.5 mL of acetic acid were transferred into a small beaker. In a separate container. 1.5 g of ferric chloride was also prepared. The chemicals were set aside for further use.

Fig 16 Ferric Chloride (left) and Acetic Acid (right)

B.2.4 Distilled Water (Chemical for Saponins Test) 10 mL of distilled water was measure and set aside for further use.

Fig. 17 10 mL Distilled Water B.2.5 Distilled Water and Ferric Chloride (Chemicals for Phenols Test) 5 mL of distilled water were transferred into a small beaker. In a separate container, 1.5g of ferric chloride was also prepared. The chemicals were set aside for further use.

Fig. 18 5 mL Distilled Water (left) and 1.5 g Ferric Chloride (right)

B.2.6 Ferric Chloride Solution (Solution for Tannin Test) 1.5 g of Ferric Chloride was weighed. Meanwhile, a volume of 10 mL of water was measured in a graduated cylinder. The ferric chloride was dissolved in water. A fraction of the solution was transferred into a test tube.

Fig. 19 1.5 g of Ferric Chloride

Fig. 20 10 mL Distilled Water

Fig. 21 Ferric Chloride Solution

B.2.7 Sodium Hydroxide Solution (Solution for Anthocyanin Test)

0.8 g of Sodium Hydroxide was weighed while a volume of 20 mL of water was measured in a graduated cylinder. The sodium hydroxide was dissolved in water.

Fig. 22 0.8 g of Sodium Hydroxide

Fig. 23 20 mL Distilled Water

Fig. 24 Sodium Hydroxide Solution

B.2.8 Nitric Acid (Chemical for Protein Test) Nitric acid was purchased and prepared and set aside for further use.

Fig. 25 Nitric acid

B.2.9Sodium Hydroxide Solution (Solution for Flavonoids Test) 0.8 g of Sodium Hydroxide was weighed while a volume of 20 mL of water was measured in a graduated cylinder. The sodium hydroxide was dissolved in water.

Fig. 26 0.8 g Sodium Hydroxide

Fig. 27 20 mL of Distilled Water

Fig. 28 Sodium Hydroxide Solution

Table1. Reagents and Solutions to be used in Phytochemical Screening Reagents/Solutions Phytochemicals

/ Chemicals Needed

Procedure in Making the Reagent 0.4 g of HgCl2 is dissolved in 15 mL of H2O and

Alkaloid

Mayer’s Reagent

poured into a solution of 1.25 KI in 2.5 mL of H2O. Sufficient H2O was added to make 25 mL 1.5 g of Naphthol was dissolved in 10 mL of

Carbohydrates

Molisch’s Reagent Acetic Acid + Ferric

ethanol. Few drops of FeCl3 was mixed with 2.5 mL of

Chloride Mixture

CH3CO2H

Distilled Water

N/A

Glycosides Saponins Phenols

Distilled Water Ferric Chloride Ferric Chloride

N/A Dissolve 13.5 g of FeCl3 in 10 mL water with 0.25

Tannins Solution Sodium Hydroxide Anthocyanin

mL of concentrated HCl. Dilute to 100 mL Dissolve 0.4 g of NaOH in 10 mL of H2O

Solution Protein

Nitric Acid

N/A

Sodium Hydroxide Flavonoids

N/A Dissolve 0.4 g of NaOH in 10 mL of H2O Solution

B.2. Phytochemical Profiling Using Color Change as Indicator Test for Alkaloids. 5 mL of Mayer’s reagent were added to 5 mL of the pure extract. Formation of yellow precipitates indicates the presence of alkaloids. Test for Carbohydrates. 2.5 mL of Molisch’s reagent were added to 5 mL of the pure extract. Formation of red or purple ring indicates presence of carbohydrates. Test for Glycosides. 2.5 mL of acetic acid and ferric chloride mixture was added to 5 mL of the pure extract. Presence of blue-green color indicates the presence of glycosides. Test for Saponins. 10 mL of distilled water was added to 5 mL of the pure extract. The mixture was shaken in a graduated cylinder for 15 minutes. Presence of layer of foam or bubbles indicates presence of saponins. Test for Phenols. 5 mL of water was added to 5 mL of the pure extract, then, ferric chloride drop was also added, formation of blue or light green color indicates presence of phenols. Test for Tannins. 5 mL of ferric chloride solution was added to 5 mL of the pure extract. Formation of dark blue or greenish-black color indicates presence of tannins. Test for Anthocyanin. 2.5 mL of Sodium hydroxide solution was added to 5 mL of the extract. Formation of blue or green precipitates indicates presence of anthocyanin. Test for Protein. 2.5 mL of nitric acid was added to 5 mL of the pure extract. Formation of yellow color indicates presence of protein. Test for Flavonoids. 5 mL of Sodium hydroxide solution was added to 5 mL of the extract. Formation of orange or intense yellow color indicates the presence of flavonoids.

Phytochemical Analysis of Snake Plant Pure Extract

I.

Alkaloids

Fig 35 Absence of Alkaloids

To test the presence of alkaloid, the Mayer’s reagent test was used. Mayer’s reagent (see figure 12) was a mixture of mercuric chloride solution and potassium iodide solution. As observed, no reaction had taken place.

II.

Carbohydrates

Fig 36 Traces of Carbohydrates Molisch’s reagent (see figure 15) was used to test the presence of carbohydrates. It is composed of naphthol dissolved in distilled water. Formation of reddish or purplish ring was the indicator that carbohydrates are present in the extract. Fig. 36 shows that reaction took place. There was a change in color. When the reagent was added to the green extract, the resulting mixture became reddish in color with areas colored with purple. There are reddish rings that are unclearly seen. Since unclearly reddish rings were formed, carbohydrates must be present in the plant. Carbohydrates play an important role in the human body. It includes sugars added during food processing and refining. Complex carbohydrates are also good sources of fiber. With the presence of carbohydrates in the plant, further studies could be employed to determine its feasibility as a fiber. As to industry, carbohydrates in the form of starch could be a component of starch-based plastics. III. Glycosides

Fig 37Absence of Glycosides Fig. 37 indicated that glycosides are not present and no reaction had taken place.

IV. Saponins

Fig 38Presence of Saponins

In testing for the presence of saponins, distilled water was added to a volume of the extract. It was shaken for fifteen minutes. To conclude the strong presence of saponins, there should be a formation of 1 cm layer of foam. In Fig. 38, it could be seen that frothy bubbles or foam was formed until one centimeter above the extract. Thus, saponins were present in the plant. Saponins, like glycosides play a vital role in foaming industry. Since both glycosides and saponins were present even glycosides were only detectable, succeeding researches could also determine the feasibility of this bioactive constituent as a foaming agent.

V. Phenols

Fig 39Traces of Phenols Fig. 39 revealed traces of phenols in the extract. Phenols are phytochemicals that could act as an antimicrobial, antidiarrheal, and antihelmintic agent. In industry, phenols are of great help to chemical processes, and to wood and plastic making. Thus, the feasibility of the plant to be a component of a bioplastic could be evaluated.

VI.Tannins

Fig 40Abscence of Tannins To test the presence or absence of tannins, ferric chloride solution was added to a volume of the extract. The color did not change that indicated the absence of tannins. Fig. 40 revealed the absence of tannins in the plant extract.

VII. Anthocyanin

Fig 41Traces of Anthocyanin The sodium hydroxide solution was used to test the presence or absence of anthocyanin. To say that it is present, the resulting reaction should make the original extract have green or white precipitates. In Fig. 41 it is clearly shown that there were precipitates formed in the extract when the sodium hydroxide solution was added. Thus, the phytochemical anthocyanin was present in the extract. Anthocyanin could be used as pH indicators to determine the acidity or basicity of a substance. Thus, further study and researches could be done to test its effectiveness as a pH indicator.

VIII.

Protein

Fig 42Presence of Protein Nitric acid was used to test the presence of protein. Formation of yellow, yellow-green or light green color is an indicator that protein is present. Fig. 42 shows that a reaction took place. There was a change in color. When the nitric acid was added to the green extract, the resulting mixture became yellowish in color with areas colored black or brown. Since, this was what happened, then, there is a protein present in the extract. Protein was actually exhibiting antimicrobial property, but since the extract did not inhibit the majority of the organisms, it can be concluded that this phytochemical has no such activity as part of the plant’s phytochemistry.

IX.Flavonoids

Fig 43Presence of Flavonoids

To test the presence of flavonoids, sodium hydroxide solution was used. The indicator of the presence of flavonoids is the formation of intense yellow color. The Papua Plant pure extract was originally green in color before NaOH solution was added. When the solution was then put into the extract, there is color change that takes place, as seen in Fig. 43. Thus, the phytochemical flavonoids was present in the extract of pure Papua Plant Leaves. Like alkaloids, flavonoids were important to antimicrobial properties.

Table 2. Phytochemical Screening of Snake Plant Leaves Pure Extract Phytochemicals Reaction (Color Change, from green pure Presence of Screened

extract of Snake Plant )

Phytochemical

Alkaloids Carbohydrates

No Reaction. +brownish Molisch’s reagent

* +

Glycosides Saponins

=purple to unclearly seen reddish rings No Reaction +colorless water + 15 minutes shaken

+++

Phenols

= formation of 1 cm frothy bubbles/foams +colorless water + orange ferric chloride

+

Tannins

=yellow green at some parts No Reaction

-

Anthocyanin

+colorless sodium hydroxide

+

=formation of green precipitates on minimal Protein

area +colorless Nitric acid

+++

Flavonoids

= intense yellow +colorless sodium hydroxide solution

+++

= intense yellow *Absent (-), Traced Amount (+), Present (++), Strongly Positive (+++)

Possible Industrial Uses of Papua Plant based on its Phytochemistry The following table shows the possible uses of Papua Plant in industry: Table 3. Possible Uses of Papua Plant (Polyscias fruticosa) based on its Phytochemistry Phytochemicals Presence Possible Uses Alkaloids + (traced amount) N/A Carbohydrates + (traced amount) Starch-based Plastics Fibers

Glycosides Saponins Phenols

-(negative) +++ (strongly positive) + (traced amount)

Tannins Anthocyanin Protein Flavonoids

-(negative) + (traced amount) +++ (strongly present) +++(strongly positive)

N/A Foaming Agents Laboratory Processes Chemical Engineering Processes Wood and Plastic Making Tanning of Leather pH Indicators Chemical Processes in food Industry Used in food industry (chocolate)

SUMMARY, CONCLUSION AND RECOMMENDATIONS Evaluation of the phytochemical constituents of plants, known as its phytochemistry, has been studied to provide basis for possible biological and industrial uses of plants. Several properties of plants have been associated to the bioactive components it has. This inducted the

researchers to arise the idea of determining the phytochemical profile of Papua Plant (Polyscias fruticosa) Leaves. In this study, extracts of Papua Plant was obtained. The fresh leaves of this plant were gathered from Bulualto, San Miguel, Bulacan. The leaves were air dried and washed with distilled water. It was cut into pieces and turned into smaller pieces using the domestic blender. On the other hand, small pieces of Papua Plants leaves were manually extracted using cheese cloth and later filtered using a filter paper for the phytochemical screening. The extract was then screened for the presence of nine phytochemicals which were alkaloids, carbohydrates, glycosides, saponins, phenols, tannins, anthocyanin, protein and flavonoids. The phytochemical profiling was conducted to determine the presence of the nine mentioned phytochemicals based on color changes after reactions took place. Reagents were prepared for the phytochemical screening. The types of observation of presence of these bioactive components were divided into four degrees. These are negative (-); traced amount (+); present (++); and strongly positive (+++). The result revealed the abundance of flavonoids, saponins and proteins. There were traced amounts of carbohydrates, alkaloids, phenols and anthocyanin. However, glycosides and tannins were absent in the plant extract. Based on the results of the phytochemical screening, properties and uses of Papua Plant extract could be predetermined. Phytochemicals have been proven to have significant relation to different biological activities and several processes in chemistry and industry. As a summary, conclusions could be enumerated as follows: 1. The phytochemistry of Papua Plant (Polyscias fruticosa) includes flavonoids, saponins and proteins (strongly positive). There were traced amounts of carbohydrates, alkaloids, phenols and anthocyanin. No traces of tannins and glycosides. 2. In industry, abundance of saponins indicated the potential of the plant to serve as foaming agents. Traced amount of phenols also suggested possible usage of Papua

Plant in wood and plastic production, as well as other chemical processes. Traced amount of carbohydrates may be a paradigm for making bioplastic using Papua Plant. Based on the findings of this investigation, it is hereby recommended that: 1. The quantitative analysis of the nine phytochemicals tested qualitatively in this study should be conducted to test exact amounts of the phytochemical components in the plant. 2. The presence of other phytochemicals such as lectins, steroids, terpenoids, etc must also be determined through phytochemical screening or other more sophisticated analysis. 3. The usage of other extracting media must be considered to determine if there are other solvents that are more compatible with Papua Plant. 4. Possible uses of Papua Plant in chemical industry should be investigated. 5. Phytochemistry of other plants, maybe of the same genus or family as that of the Papua Plant should also be analyzed.

BIBLIOGRAPHY Related Studies “On The Antipyretic, Anti-Inflammatory, Analgesic and Molluscicidal Properties Of Polyscias Fruticosa (L) Harms” Bensita Mary Bernard, Nilani Pakianathan, and Madhu C. Divakar “A Pharmacognostic Report on the Leaf and Root of Polyscias Fruticosa (L.) Harms”

Bensita Mary Bernard, Nilani Pakianathan, R. Venkataswamy, And Madhu C. Divakar Studies On The Adaptogenic and Antibacterial Properties Of Polyscias Fructicosa (L) Harms M.B. Bensita, P.Nilani, S. Sandhya M Pharmacognosy And Photochemistry Laboratory, College Of Pharmacy, Sripms, Coimbatore, Tamil Nadu – 641 04. Received: 8th February, 1998 Accepted: 11th August, 1998 DIURETIC ACTIVITY OF Polyscias Fruticosa (L.) HARMS *R. Varadharajan and D. Rajalingam *Kamalakshi Pandurangan College of Pharmacy, Ayyampalayam, Tiruvannamalai, Tamil Nadu - 606 603. The Leaf Volatile Oil of Polyscias Fruticosa (L.) Harm Oliveros-Belardo, Luz ET. Al. http://www.plantneurobiology.org/phytochemistry.php http://wiki.answers.com/Q/What_is_phytochemical_screening http://www.plantneurobiology.org/phytochemistry.php http://www.tjclarkminerals.com/phytochemicals/phytochemicals.htm http://en.wikipedia.org/wiki/phytochemicals http://www.friedli.com/herbs/phytochem/alkaloids/alkaloid1.html http://www.britannica.com/EBchecked/topic/15672/alkaloid http://www.madsci.org/FAQs/anthocyanins.html http://en.wikipedia.org/wiki/anthocyanin http://www2.chemistry.msu.edu/faculty/reusch/VirtTxtJml/carbhyd.htm http://www.nlm.nih.gov/medlineplus/carbohydrates.html http://medical-dictionary.thefreedictionary.com/glycoside http://en.wikipedia.org/wiki/Glycoside http://en.wikipedia.org/wiki/Phenols http://www.thefreedictionary.com/phenols http://en.wikipedia.org/wiki/Proteins http://www.thefreedictionary.com/protein http://www.phytochemicals.info/phytochemicals/saponins.php http://www.herbs2000.com/h_menu/saponins.htm http://www.britannica.com/EBchecked/topic/582701/tannin http://en.wikipedia.org/wiki/Tannin

APPENDICES

APPENDIX A Structural features and activities of various phytochemicals from plants Phytochemical s Phenols and Polyphenols

Quinones

Structural features

Example(s)

Activities

C3 side chain, - OH groups, phenol ring

Catechol, Epicatechin, Cinnamic acid

Antimicrobial, Anthelmintic, Antidiarrhoeal

Aromatic rings, two ketone substitutions

Hypericin

Antimicrobial

Abyssinone Chrysin, Quercetin, Rutin Totarol

Antimicrobial Antidiarrhoeal

Antimicrobial, Anthelmintic, Antidiarrhoeal Antimicrobial

Flavones Flavonoids Flavonols

Phenolic structure, one carbonyl group Hydroxylated phenols, C6-C3 unit linked to an aromatic ring Flavones + 3-hydroxyl group

Tannins

Polymeric phenols (Mol. Wt. 500-3000)

Ellagitannin

Coumarins

Phenols made of fused benzene and αpyrone rings

Warfarin

Terpenoids and essential oils

Acetate units + fatty acids, extensive branching and cyclized

Capsaicin

Antimicrobial Antidiarrhoeal

Alkaloids

Heterocyclic nitrogen compounds

Berberine, Piperine, Palmatine, Tetrahydropalmatine

Antimicrobial, Anthelmintic, Antidiarrhoeal

Lectins and Polypeptides

Proteins , Mannose-specific agglutinin Sugar + non carbohydrate moiety Amphipathic glycosides

Fabatin

Antimicrobial

Amygdalin

Antidiarrhoeal

Vina-ginsenosides-R5 and -R6

Antidiarrhoeal

Glycosides Saponins

APPENDIX B Mechanism of action of some phytochemicals Phytochemicals Quinones

Activity Antimicrobial

Mechanism of action Binds to adhesins, complex with cell wall, inactivates enzymes

Flavonoids

Antimicrobial

Complex with cell wall, binds to adhesins

Antidiarrhoeal

Inhibits release of autocoids and prostaglandins, Inhibits contractions caused by spasmogens, Stimulates normalization of the deranged water transport across the mucosal cells,

Polyphenols and

Antimicrobial

Inhibits GI release of acetylcholine Binds to adhesins, enzyme inhibition, substrate

Tannins

Antidiarrhoeal

deprivation, complex with cell wall, membrane

Anthelmintic

disruption, metal ion complexation Makes intestinal mucosa more resistant and reduces secretion, stimulates normalization of deranged water transport across the mucosal cells and reduction of the intestinal transit, blocks the binding of B subunit of heat-labile enterotoxin to GM1, resulting in the suppression of heat-labile enterotoxin-induced diarrhea, astringent action Increases supply of digestible proteins by animals by forming protein complexes in rumen, interferes with energy generation by uncoupling oxidative phosphorylation, causes a decrease in G.I.

Coumarins Terpenoids and

Antiviral Interaction Antimicrobial

metabolism with eucaryotic DNA Membrane disruption

essential oils

Antidiarrhoeal

Inhibits release of autocoids and prostaglandins

Alkaloids

Antimicrobial

Intercalates into cell wall and DNA of parasites

Antidiarrhoeal

Inhibits release of autocoids and prostaglandins

Anthelmintic

Possess anti-oxidating effects, thus reduces nitrate generation which is useful for protein synthesis, suppresses transfer of sucrose from stomach to small intestine, diminishing the support of glucose to the helminthes, acts on CNS causing

Lectins and Polypeptides Glycosides Saponins

Steroids

Antiviral

paralysis Blocks viral fusion or adsorption, forms disulfide

Antidiarrhoeal Antidiarrhoeal

bridges Inhibits release of autocoids and prostaglandins Inhibits histamine release in vitro

Anticancer

Possesses membrane permeabilizing properties

Anthelmintic

Leads to vacuolization and disintegration of

Antidiarrhoeal

teguments Enhance intestinal absorption of Na+and water