Standardization and Preliminary Phytochemical Investigation of the Fruits of Grewia asiatica Linn.

Hayat M. Mukhtar*, Harjas Kaur, Sandeep Singh, Manwinder Singh

Natural Product Research Laboratory, Department of Pharmacognosy and Phytochemistry, Shaheed Bhagat Singh College of Pharmacy, Patti, Amritsar, Punjab. 143 416.

ABSTRACT:

Grewia asiatica is a small deciduous tree consisting of ripe fruits, seeds, flowers, bark and leaves of the plant belonging to family Tiliaceae. In present study, various standardization parameters of fruits like macroscopical, microscopical characters, physicochemical parameters like total ash (5.16%), water soluble ash (2.5%), acid insoluble ash (1%), sulphated ash (0.66%), loss on drying (8.3%), swelling index (0.1), foaming index (less than 100), successive extractive values in various solvents like petroleum ether (1.6%), benzene (2.1%), chloroform (2.6%), ethyl acetate (3.7%), methanol (30.9%). Cold Maceration values, petroleum ether (3%), benzene (3.3%), chloroform (4%), ethyl acetate (4.3%), methanol (45.6%). Behaviour on treatment with different chemical reagents of the dried fruits, fluorescence characters under visible and ultralviolet light were studied to fix some pharmacognostic parameters that vary from region to region. Preliminary phytochemical studies on different extracts were reported.

KEYWORDS: Grewia asiatica, solvent extracts, UV chamber, Tiliaceae, fruits.

INTRODUCTION:

It is a genus of about one hundred and fifty species and forty-two species are native to India. It is the only genus in the family that yields edible fruits. It is commonly known as Phalsa. It is a small deciduous tree up to 7m tall with grey rough bark. The branches are long, slender and drooping. The fruit is a globose drupe that is 1.8-2.2 cm in diameter. When ripe it is red or purple, finely warty and with stellate hairs. The soft flesh is greenish white and stained with purplish red¹. Plant is reported to possess radioprotective², antibacterial³, hepatoprotective⁴, antipyretic⁵, antiemetic⁶ and antidiabetic⁷ activity. The fruit is astringent, cooling, stomachic and used in heart and blood disorders and fevers. The fruits are aphrodisiac; they allay thirst and burning sensations, remove biliousness, cure inflammation⁸. An infusion of the bark of the plant is used as demulcent. The sandals are reported to use the root bark for rheumatism. The leaf extract is applied to pustular eruptions⁹. The fruit is also good against throat trouble. The leaves are also used medicinally, chiefly for external applications¹⁰.

MATERIAL AND METHODS:

Plant material:

The Fruits of Grewia asiatica were collected from Amritsar district of Punjab in the month of June. The plant was identified and authenticated by botanist, Dr. Adarash Pal Vig, Department of Botanical and Environmental Sciences, Guru Nanak Dev University, Amritsar. Its voucher number (1260) specimen was deposited in the same department for reference.

Processing of Plant material:

After authentication, fruits were dried at room temperature until they were free from the moisture and subjected to physical evaluation for different parameters.

Reagents:

All the reagents were of Analytical grade and purchased from S.D Fine- Chem. Ltd., Mumbai, India.

Table 1: Organoleptic characters of the fruits of Grewia asiatica

Organoleptic Characters	Observations
Type	Simple
Colour	Red or dark purple
Odour	Pleasant
Taste	Acidic
Size	1.8-2.2 cm in diameter
Shape	Globose

Table 2. Physicochemical parameters of fruits of Grewia asiatica

Parameters	Values obtained (% w/w)
Total ash	5.16
Acid insoluble ash	1
Water soluble ash	2.5
Sulphated ash	0.66
Swelling index	0.1
Foaming index	Less than 100
Loss on drying	8.3

Table 3. Extractive values of fruits of Grewia asiatica

Solvents	Successive extractive value (% w/w)	Cold maceration value (% w/w)
Petroleum ether	1.6	3
Benzene	2.1	3.3
Chloroform	2.6	4
Ethyl acetate	3.7	4.3
Methanol	30.9	45.6

Methods:

The organoleptic characters including colour, odour, taste and external features of fruits of Grewia asiatica were observed and the results were recorded in Table1. The extractive values were determined by successively starting from petroleum ether, benzene, chloroform, ethyl acetate and methanol by using soxhlet extraction apparatus. The extractive values were obtained after evaporation of solvent under reduced pressure using vaccum rotary evaporator. The behaviour of the powdered fruits with different chemical reagents were studied and fluorescence characters were also observed under ultraviolet and visible light (compared with Nerolac Synthetic and Puja Satin Enamel Colour Reference Card). Preliminary phytochemical tests of different extracts were performed with specific reagents^11,12.

RESULTS AND DISCUSSION:

The dried fruits of Grewia asiatica were subjected to standard procedures according to WHO Guidelines for the determination of various physicochemical parameters. The following parameters were determined:

Ash Values:

The determination of ash value is meant for detecting low-grade drugs, exhausted drugs, sandy or earthy matter. The results of ash values were recorded in Table 2.

Extractive Values:

This method determines the amount of active constituents extracted with solvents from a given amount of medicinal plant material. It is employed for materials for which as yet no suitable chemical or biological assay exists. The air dried, accurately weighed drug was treated with solvents: petroleum ether, benzene, chloroform, ethyl acetate and methanol. The values were recorded in Table 3.

Fluorescence analysis:

The powdered drug was examined under ordinary light, short UV (254 nm) and long UV (366 nm) light using different solvents and compared with Nerolac Synthetic and Puja Satin Enamel Colour Reference Card. The results were recorded in Table 4.

Table 4. Fluoresence analysis of powder of fruits of Grewia asiatica with various chemical reagents under visible light, short and long wave length* .

Solvents	Visible light	UV (254 nm)	UV(366 nm)
Drug Powder	Pumpernickel 2824	Steel grey	Brown
Powder drug+ conc. HCl	Cherry	Espresso 2847	Brown
Conc.HCl +H2O	Leaf brown	Espresso	Brown
Conc. HNO3	Golden brown	Cherry	Brown
Conc. HNO3+ H2O	El greco bronze 2063	Mint green	Brown
Picric acid	Cinnamon tea	Olive green	Brown
Picric acid+ H2O	Mid buff	Olive green	Brown
Ethyl acetate	Golden brown	Safe grey	Brown
Methanol	Espresso 2847	Deep green	Brown
Ethanol	Pebble 2787	Bufftone	Brown
Chloroform	Cherry	Espresso 2847	Brown
Pet. Ether	Cherry	Espresso 2847	Brown
Distilled water	Light Biscuit 6023	Cherry	Brown
10% NaOH	Cherry	Espresso 2847	Brown
Iodine	Espresso 2847	Deep green	Brown

*Compared with Kansai Nerolac synthetic and Puja Satin enamel a subsidiary of Kansai Paint Co. Ltd., Japan, S-2011

Table 5. Preliminary Phytochemical screening of fruits of Grewia asiatica

Phyto constituents	Petroleum ether extract	Benzene extract	Chloroform extract	Ethyl acetate extract	Methanol extract
Alkaloids	-	-	+	+	+
Glycosides	-	-	-	-	+
Proteins and Amino acids	+++	+++	+++	+++	+++
Carbohydrates	-	-	-	+	+
Tannins	+++	+++	+++	+++	+++
Fats and Fixed oils	+	+	-	-	+
Acids	+	-	+	-	-
Saponins	+	+	+	-	-
Steroids	-	-	-	-	-
Gums and mucilages	+	+	+	+	+

Table 6: Behavioural analysis of powdered fruits of Grewia asiatica with various chemical reagents.

TREATMENT WITH CHEMICALS	OBSERVATIONS
Powder	Brown
Powder + conc. HCl	Leaf brown
Powder + conc. H2SO4	Espresso 2847
Powder + conc.HNO3	Reddish brown
Powder + Acetic acid	Greenish brown
Powder + Picric acid	Golden brown
Powder + iodine solution	Yellowish brown
Powder + 10% NaOH	Brown
Powder + FeCl3	Brown
Powder + Methanol	Brown
Powder + Ethanol	Espresso
Powder + Ammonia solution	Brown
Powder + Chloroform	Cherry

Phytochemical screening:

The plant material was subjected to preliminary phytochemical screening for the detection of various plant constituents. The extracts obtained from successive solvent extraction were subjected to qualitative test for the identification of various plant constituents like alkaloids, carbohydrates, glycosides, proteins and amino acids, saponins, steroids, acidic compounds, mucilage and fixed oils and fats¹³. The results were recorded in Table 5.

Powdered drug reaction with various chemical reagents:

The powdered drug was treated with various chemicals like concentrated hydrochloric acid, sulphuric acid, nitric acid, 10% NaoH, picric acid, iodine solution, methanol, ethanol, acetic acid, chloroform, petroleum ether, ferric chloride, ammonia solution. The behavioural change of the drug was determined¹⁴. The results were recorded in Table 6.

CONCLUSION:

Generated data can be used for determining correct identity and purity of plant parts and for the detection of adulteration. Botanical authentication and physicochemical parameters will give an idea about the quality of drug. All these parameters which are being reported could be useful in identification of distinctive features of the drug. From the preliminary phytochemical study, it was concluded that only selected species of Grewia asiatica Linn. contained the reported phytoconstituents. Hence, detailed screening may be done to isolate the active constituents so that it may be scientifically proved to access the pharmacological responses of the plant to ascertain its folklore uses.

ACKNOWLEDGEMENT:

The authors wish to thank SBS College of Pharmacy, Patti, providing necessary facilities for carrying out the research.

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Received on 11.05.2012

Modified on 24.05.2012

Accepted on 30.05.2012

© A&V Publication all right reserved

Research Journal of Pharmacognosy and Phytochemistry. 4(4): July- August 2012, 212-214