INTRODUCTION:

Tamarind is the ripe fruit of the Tamarindus indica tree, which is used as a condiment, or more precisely as an ‘acidulant’ like amchur. Each and every part of the Tamarind tree, especially the fruit is beneficial for the society. The sweetish acidic pulp of the fruit is a product of commercial importance. Pulp of Tamarind fruit is an important component in chutneys, pickles, jams curries, sauces, ice cream, sharbat and “tamarind fish”, a special Indian seafood pickle. Tamarind is extensively used in the Indian system of medicine, Ayurveda. Tamarind preparations are universally recognized as refrigerants in fevers and as laxatives .The fruit pulp is used as digestive, a remedy in bile disorders, to alleviate sunstroke, in datura poisoning and alcoholic intoxication.

Tamarind drink is popular all over the world. The Tamarind has recently become popular as bonsai, in Indonesia, Taiwan and Philippines. The seeds are sometimes used by children in traditional board games such as Chinese checkers(China), Dhakon (Java) etc. Tamarind belonging to family Caesalpiniaceae is scientifically known as Tamarindus indica Linn a perennial evergreen tree with a spreading crown; feathery evergreen foliage and fragrant flowers. It grows widely in tropical and subtropical regions, yielding hard yellowish wood and long pods with edible chocolate-colored acidic pulp. It is cultivated almost in all the states of the country, except in Himalayas and western dry regions. Tamarind is not only a food item but its pulp, leaves and bark also medical applications. Tamarind fruit pulp has been an important culinary ingredient in India for a very long time. It is extensively used in Tamil Naidu, Karnataka and Andhra Pradesh cuisines, particularly in the preparation of Rassam, Sambhar, Vatthakuzhambu and Puliyogare. Tamarind trees provide shade on the country roads and severe also as ornamental trees^[1].

Medicinal Uses

· Laxative and constipation reliever(Ripe and Unripe fruit pulp, mixed with milk, honey or lemon juice, due to high amount of maleic acid, tartaric acid and potassium acid tartarate)

· Wound healer (Leaves and bark ,due to high amount of tannin, applied externally on the spot as a decoction or as powder)

· Antipyretic (Fresh Fruits)

· Antimalarial agent (Tamarind fruit pulp and leaves)

· Aphrodisiac (Different plant parts i.e. flowers, leaves, bark and fruit pulp)

· Abdominal pain reliever (Fresh bark or stem)

· Mouthwash

· Anti-inflammatory (Leaves and Bark ,due to high tannin content)

· Preservative (Fruit)

· Anthelmintic (Bark)

· Antibacterial (Fruit)

· Antidiabetic (Leaves)

· Antihypertensive

· Antiemetic

· Lotion and poultice of fresh leaves is applied to swellings, boils, cuts for relieving pain and inflammation^[2].

MATERIALS AND METHODS:

Collection and Authentication of Tamarindus indica Plant Seeds

The seeds of Tamarindus indica (IMLI) were collected from the Sandhu Nursery, Amritsar and authenticated by Dr. Adarshpal Vig (Dept. of Botanical and Environmental Science Guru Nanak Dev University Amritsar, Punjab), Voucher No:-1704 .

Plant Profile

Botanical Information

Botanical Name : Tamarindus indica

Family: Caesalpiniaceae

Genus : Tamarindus L.

Species : Tamarindus indica

Common Name : Imli

Vernacular Names

English : Tamarindus

Sanskrit: Tintiri

Hindi : Imlii

Bengali : Ambli

Marathi : Ambali

Telgu : Chinta

Kannada: Amla

Ladakh : Ambli ,Amli

Punjab: Imbli

Oryia : Kaniya ,Omlika

Parts used: Flower, Leaves, Root and Whole plant

Figure No.-1 Seeds and Fruit

Figure No.-2 Tamarindus Indica (IMLI Tree)

Morphology

Tamarindus indica is a large evergreen tree up to 30 m tall, bole usually 1-2 m, up to 2 m diameter; crown dense, widely spreading, rounded; bark rough, fissured, greyish-brown. Leaves alternate, compound, with 10-18 pairs of opposite leaflets; leaflets narrowly oblong, 12-32 x 3-11 mm, petiole and rachis finely haired, midrib and net veining more or less conspicuous on both surfaces; apex rounded to almost square, slightly notched; base rounded, asymmetric, with a tuft of yellow hairs; margin entire, fringed with fine hairs. Stipules present, falling very early. Flowers attractive pale yellow or pinkish, in small, lax spikes about 2.5 cm in width. Flower buds completely enclosed by 2 bracteoles, which fall very early; sepals 4, petals 5, the upper 3 well developed, the lower 2 minute. Fruit a pod, indehiscent, subcylindrical, 10-18 x 4 cm, straight or curved, velvety, rusty-brown; the shell of the pod is brittle and the seeds are embedded in a sticky edible pulp. Seeds 3-10, approximately 1.6 cm long, irregularly shaped, testa hard, shiny and smooth. As the dark brown pulp made from the fruit resembles dried dates, the Arabs called it ‘tamar-u’l-Hind’, meaning ‘date of India’, and this inspired Linnaeus when he named the tree in the 18th century. Tamarindus is a monospecific genus^[3].

Phytochemical Studies

1. Extraction

Phytochemicals are the chemical substance of organic nature, which are formed in plants through the activity of their individual cells for serving different purposes. The medicinal value of any plant drug, however, depends on the nature of the chemical constituents in it and is referred to as active principle.

1.1 Preparation of plant Extracts

The plant leaves were shade dried and were powdered, the coarse powder was subjected to extraction with petroleum ether and alcohol (70%) individually in Soxhlet apparatus and also drug powder was subjected to aqueous extraction.

1.1.1 Petroleum Ether Extract

The Powdered seeds weighing 1kg (1000g) were packed in 5000ml flat bottom and added 4 liters of petroleum ether and kept for 1 week for cold maceration with intermittent shaking. After that the plant extract was filtered and concentrated under vacuum with the help of distillation assembly.

1.2.2 Aqueous Extract

The Powdered seeds weighing 1kg (1000g) were packed in 5000ml flat bottom and added 4 liters of distilled water and 10ml chloroform as preservative and kept for 1 week for cold maceration with intermittent shaking. After that the plant extract was filtered and concentrated under vacuum with the help of distillation assembly.

1.1.2 Ethanol Extract

The Powdered seeds weighing 1kg (1000g) were packed in 5000ml flat bottom and added 4 liters of ethanol and kept for 1 week for cold maceration with intermittent shaking. After that the plant extract was filtered and concentrated under vacuum with the help of distillation assembly.

Table No: 1 The nature and extractive values of seed extracts of Tamarindus indica (Pet ether, ethanolic, Aqueous extract of Tamarindus indica)

S. No.	Extracts	Colour	Yield (%w/w)
1.	PETI	Yellowish	4.4
2.	EETI	Yellowish somewhat sticky	9.2
3.	AETI	Yellowish, bit sticky	15.3

1.2 Qualitative phytochemical analysis

The petroleum ether,aqueous and ethanol extract of the seeds of Tamarindus Indica was subjected to various chemical tests for identification of its different active constituents and the results were recorded in Table No.-2.

1.2.1 Detection of carbohydrates

Extracts were dissolved individually in 5 ml of distilled water and filtered. The filtrates were used to test the presence of carbohydrates.

(A) Molisch’s Test

Filtrates were treated with 2 drops of alcoholic a- naphthol solution in a test tube and 2ml conc. Sulphuric acid were added carefully along the sides of the test tubes. Formation of violet ring at the junction indicates the presence of carbohydrates.

(B) Benedict’s Test

Filtrates were treated with benedict’s reagent and heated on water bath. Formation of an orange red precipitate indicated the presence of reducing sugars.

(C) Fehling’s Test

Filters were hydrolyzed with dilute hydrochloric acids, neutralized with alkali and heated with Fehling’s A and B solutions. Formation of red precipitate indicates the presence of reducing sugars.

1.2.2 Detection of alkaloids

(A) Mayer’s Test: Alkaloids give cream colour precipitate with Mayer's reagent (potassium mercuric iodide solution).

(B) Dragandroff’s Test: Alkaloids give reddish brown precipitate with Dragandroff’s reagent (potassium bismuth iodide solution).

(C) Wagner's Test: Alkaloids give a reddish brown precipitate with Wagner's reagent (Solution of iodine in potassium iodide).

(D) Hager's Test: Alkaloids give yellow colour precipitate with Hager's reagent (saturated solution of picric acid).

1.2.3 Detection of glycosides

Extracts were hydrolyzed with dilute hydrochloric acid, and the hydrolysate was subjected to glycosides tests.

(A) Modified Borntrager's Test: Boiled 200 mg of the test material with 2 ml of dilute sulphuric acid, 2 ml of 5% aqueous ferric chloride solution for 5 min and continued the test as above. As some plant contain anthracene aglycone in a reduced form, if ferric chloride was used during the extraction, oxidation to anthraquinones took place, which showed response to the Borntrager's test.

(B) Legal’s Test: The extracts were treated with sodium nitro prusside in pyridine and methanolic alkali.The formation of pink to red colour indicate the presence of cardiac glycoside.

(C) Keller Killiani Test (Test for deoxy sugars):

Extracted the drug with chloroform and evaporated it to dryness, added 0.6 ml of glacial acetic acid containing a trace amount of ferric chloride. Transferred to a small test tube; added carefully 0.5 ml of concentrated sulphuric acid by the side of the test tube, blue color appears in the acetic acid layer if cardiac glycoside was present.

1.2.4 Detection of saponins

(A) Froth Test: Placed 1 ml solution of drug in water in a semi micro tube shake well and note the stable froth.

(B) Libermann Buchard Test: Extract treated with few drops of acetic anhydride, boil and cool, cone. Sulphuric acid is added from the sides of the test tube, shows a brown ring at the junction of two layers and the upper layer turns green which shows the presence of steroids and formation of deep red colour indicates the presence of triterpenoids.

1.2.5 Detection of phytosterols

(a) Salkowski Test: Treat extract in chloroform with few drops of conc. sulphuric acid, shake well and allow to stand for some time, red colour appears at the lower layer indicates the presence of steroids and formation of yellow coloured lower layer indicates the presence of triterpenoids.

1.2.6 Detection of fixed oils and fats

(a) Soap Test: The extracts were heated on water bath with 0.5 N alcoholic potassium hydroxide solutions. Formation of soap indicates the presence of fixed oils and fats.

1.2.7 Detection of Phenolic compounds and Tannins

(A) Gelatin Test: Extract with 1% gelatin solution containing 10% sodium chloride gives white precipitate.

(B) Ferric chloride Test: Test solution gives blue green color with ferric chloride.

(C) Vanillin hydrochloride Test: Test solution when treated with few drops of vanillin hydrochloride reagent gives purplish red color.

(D) Shinoda Test: To the test solution add few fragments of magnesium ribbon and add conc. hydrochloric acid drop wise, pink scarlet, crimson red or occasionally green to blue colour appears after few min.

(E) Alkaline Reagent Test: To the test solution add few drops of sodium hydroxide solution; formation of an intense yellow colour, which turns to colourless on addition of few drops of dil. acid, indicates presence of flavonoids.

1.2.8 Detection of proteins and amino acids

(A) Millons Test: Test solution with 2 ml of Millons reagent (mercuric nitrate in nitric acid containing traces of nitrous acid), white precipitate appears, which turns red upon gentle heating.

(B) Ninhydrin Test: Amino acids and proteins when boiled with 0.2% solution of ninhydrin (Indane 1, 2, 3 trione hydrate), violet colour appears.

(C) Biuret Test: The extracts were treated with 1ml of 10% sodium hydroxide solution and heated . A drop of 0.7% copper sulphate solution to the above mixtures was added . The formation of purplish violet color indicates the presence of proteins^[4].

Table No. 2 Data Showing Results of Chemical Tests For Ethanol Extract, Aqueous extract and Pet. Ether extract of Tamarindus indica:-

Tests

Ethanol extract

Aqueous extract

Pet ether extract

Alkaloids

a. Dragondroff’s Test

b. Wagner’s Test

c. Mayer’s Test

d. Hager’s Test

Carbohydrates

a. Molish’s Test

b. Fehling’s Test

c. Benedict’s Test

Proteins

a. Biuret Test

b. Xanthoprotein Test

c. Lead Acetate Test

Amino Acid

a. Ninhydrin Test

Glycoside

a. Legal Test

b. Baljet Test

Steroids and Sterols

a. Liberman Burchard Test

b. Salkowsky Test

Anthraquinones

a. Borntrager’s Test

Flavonoids

a. Extract+Tin+HCl

Tannins and Phenols

Triterpenoids

Saponin Test

a. Foam Test

Fixed oils

a. Spot Test

Note: (+) sign indicates Presence (-) sign indicates Absence

RESULT:

The results of the extractive values and phytochemical screening of different extracts of Tamarindus indica seeds are shown in Table-1. Seed extract of Tamarindus Indica were selected for the present study. About 500g of dried seeds were powered and extracted with Ethanol, Aqueous and Pet Ether Extract. Various chemical tests were performed like detection of Alkaloids, Flavonoids, Carbohydrates, Proteins, Amino Acids, Glycosides, Steroids, Sterols, Anthraquinones, Tannins, Phenols, Triterpernoids, Saponins and Fixed Oils .The test for Alkaloids was negative for all the three extracts. The test for Carbohydrates was positive for all the three extracts. Protein test were also positive for all the three extracts. The test for Amino Acids was positive for all the three extracts. The test for Steroids and Sterols was positive for ethanol extract and negative for aqueous and petroleum ether extract . The test for Anthraquinones was negative for all the three extracts. The phytochemical analysis was carried out on ethanolic seed extract of Tamarindus indica showed the presence of some bioactive compounds in the plant, but alkaloids and gylcosides are totally absent in the ethanolic, pet ether and water extract of plant. Most of chemical constituents are present in ethanolic extract of Tamarindus indica Shown in (Table-1). Evaluation of phytochemical study further help in evaluation of pharmacological activity depending upon chemical constituents present in the plant seeds extracts.

DISCUSSION:

The various qualitative tests indicate that the seeds of Tamarindus indica contain Steroids, Sterols, Tannins, Phenols, Triterpernoids , Proteins, and Fixed Oils present in aqueous extract and ethanolic extract. Different extract of Tamarindus indica plant were subjected to phytochemical screening for evaluation of chemical constituents. Phytochemical screening revealed the presence of carbohydrates, proteins, saponins, tannins, triterpenoids in ethanol, pet ether and water extract of plant. The plant Tamarindus indica locally known as Tetul belongs to the family Fabaceae. The plant is extensively grown in all over the Bangladesh, is widely used all over Tropical Africa, Sudan, India, Pakistan for different purposes. Different parts of this plant are used in the indigenous systems of medicine for the treatment of a variety of human ailments^[5],[6]. Tamarindus indica is widely used in traditional medicine in Africa for the treatment of many diseases such as fever, dysentery, jaundice, gonococci and gastrointestinal disorders^[7].. Pharmacological investigations on Tamarindus indica extracts reported them to have antibacterial, antifungal hypoglycaemic, cholesterolemic cytotoxic effects etc^[8]. Plants have the capacity to synthesize a diverse array of chemicals and understanding how phytochemicals function in plants may further our understanding of the mechanisms by which they benefit humans. Evaluation of photochemical further help in evaluation of pharmacological activity. From the present study we revealed that carbohydrates, proteins, tannins sapponins were present in the ethanolic, water and pet ether extracts of seeds of plant.

REFERENCES:

1 Chopra RN, Nayar SL, Chopra IC. Glossary of Indian Medicinal Plants. J. Bio. Sci. Vol-1;1958; 75-78

2. Ibrahim E and Abbas SA. 1995 Chemical and biological evaluation of Tamarindus indica L. growing in Sudan. ActaHortic. J. Bio.SciVol.(1); 42–59.

3. Coutino RR, Hernandez CP, Gillis RH. Lectins in fruits having gastro-intestinal activity J Ethanopharmacol (2); 2001; 271–77.

4. Khandelwal KR. Practical phramcognosy techniques and experiments, 19th edition. Nirali prakashan : 149-153.

5. Nikkon F, Saud ZA, Rahman H. and Haque ME. In vitro antibacterial activity of the compound Isolated from Chloroform Extract of Moringa oleifera Lam. 6(22); 2003; 1888-1890.

6. Rahman, M.M., Wahed, M.I.I., Biswas, M.H., Sadik, G.M. and Haque, M.E. In vitro antibacterial activity of the compounds of Trapa bispinosa Roxb. Science. 1; 2001; 214-216.

7. Ferrara, L. Antioxidant activity of Tamarindus indica L., Ingredient Alimentary. 4(6); 2005; 13-15.

8. Khanzada SK, Shaikh W, Sofia W, Kazi T, Usmanghani K, Kabir A and Sheerazi TH. Chemical constituents of Tamarindus indica. Medicinal plant in sindh; Pak. J. Bot., 40(6); 2008; 2553-2559.

Received on 22.03.2014 Modified on 28.03.2014

Res. J. Pharmacognosy & Phytochem. 6(2): April-June 2014; Page 75-79

Medicinal Uses

Phytochemical Studies

1. Extraction

1.1 Preparation of plant Extracts

1.1.1 Petroleum Ether Extract

1.1.2 Ethanol Extract

Table No: 1 The nature and extractive values of seed extracts of Tamarindus indica (Pet ether, ethanolic, Aqueous extract of Tamarindus indica)

S. No.

Extracts

Colour

Yield (%w/w)