Physicochemical and Preliminary Phytochemical Studies on the Fruit of Tribulus terrestries Linn.

A.K. Meena¹, Kiran Sharma²*, Vikas Jain², Bhavana Pal², AjitK.¹, Uttam Singh², R. Singh and M.M. Rao¹

¹National Institute of Ayurvedic Pharmaceutical Research, Patiala– 47001, Punjab, (India).

²Shobhit University, Meerut, UP

ABSTRACT:

The present communication attempts to evaluate the physicochemical and preliminary phytochemical studies on the fruit of Tribulus terrestries Linn. Zygophyllaceae family.

Tribulus terrestries is an annual or perennial, prostrate herb with many slender, spreading branches and silky-villous young parts. The plant grows wild throughout India, the shrub thrives in well irrigated black soil upto attitudes of 3000m. Tribulus terrestries is traditionally used in the formulation for antiurolithiatic activity, anthelmintic activity, anti-inflammatory activity, cardiac activity, aphrodisiac activity etc.

As there is no detailed standardisation work reported on fruit, the physicochemical parameters, preliminary phytochemical constants, toxic heavy metals, pesticide residue, and aflatoxin analysis are carried out. The study revealed specific identities for the particular crude drug which will be useful in identification and control to adulterations of the raw drug.

KEYWORDS: Extractive values, Ayurvedic drug, Toxic metals, physicochemical studies,

INTRODUCTION:

Since origin of human’s life, plants continue to play a curative and therapeutic role in preserving human health against disease and decay. The widespread use of herbal remedies and healthcare preparations, such as those described in ancient texts like the Vedas and the Bible have been traced to the occurrence of natural products with medicinal properties^1,2. In this context, India being a subtropical country is a good repository of plants that are widely used in the preparation of herbal therapies.

Tribulus terrestries Linn. is also known as Land-caltrops, Puncture-vine, Gokharu in Hindi, Negalu, Sennanegalu in Kannada, Nerinnil in Malayalam, Nerinci in Tamil, Goksurah in Sanskrit, Palleru, Cinnapalleru in Telugu.

Tribulus terrestries is an annual or perennial, prostrate herb with many slender, spreading branches and silky-villous young parts. Leaves are abruptly simple, pinnate and opposite. Leaflets almost sessile, rounded or oblique at the base, mucronate at the apex, flowers bright yellow, solitary, pseudo axillary or leaf opposed. Fruits are 5 angled or winged spinous tuberculate woody schizocarp, separating into five cocci, each coccus having two long, stiff, sharp divaricate spines towards the distal half and two shorter ones nearer the base, seeds one or more in each coccus. The plant grows wild throughout India, the shrub thrives in well irrigated black soil up to attitudes of 3000m ^3-4.

The roots and fruits of Tribulus terrestries Linn. are sweet, cooling, antiurolithiatic activity⁵, aphrodisiac⁶, emollient, appetiser, digestive, anthelmintic, expectorant, anodyne, anti-inflammatory, alterant, laxative, cardiotonic⁷, styptic, lithotriptic and tonic. They are useful in gonorrhoea, inflammation, menorrhagia, strangury, leprosy, skin diseases, verminosis and general weakness^8-10.

The ash of the whole plant is good for external application in rheumatic-arthritis^11-12. The diuretic properties of the plant are due to the large quantities of the nitrates present as well as the essential oil which occurs in the seeds¹³.

The major constituents of this plants are steroidal saponins1 namely terrestrosins A, B, C, D and E, desgalactotigonis, F-gitonis, desglucolanatigoneis, gitnin etc., which on hydrolysis yield jdiosgenis, hecogenis and neotigogenin^14-17 etc. There are other minor constituents like alkaloids (uncharacterised)¹⁸, common phyto sterols namely, b-sitosterol, stigmasterol, a cinnamic amide derivative - terrestiamide and 7-methylhydroisdamone.

In Ayurvedic system of medicine Tribulus terrestries is used in treatment of Dysuria, polyuria, oedema, bronchial asthma, piles cardiac diseases, urinary calculi, anorexia, diseases of nervous system. In Siddha system of medicine Nerunjil is used in oliguria, fever, urinary calculi, diseases of vatam, leucorrhoea, thirst, retention of urine, aphrodisiac¹⁹.

Tribulus terrestries Linn. has threats due, to harvest for medicines loss of habitat and trade. Therefore, the present paper attempts to evaluate the physicochemical parameters, preliminary phytochemical screening and heavy metal analysis of the fruits of Tribulus terrestries Linn. for identification of the drug in dry form and control the adulterants.

RESULT AND DISCUSSION:

The fruits of Tribulus terrestries Linn. was collected and analysed the various standardisation parameters. Preliminary phytochemical results showed the presence or absence of certain phytochemicals in the drug. The tests performed using n-Hexane, Chloroform, Ethyl acetate, alcoholic and water extracts. Phytochemical test revealed the presence, Alkaloid, steroidal glycoside, saponins, flavonoids, polysaccharides, Tannin and results are given in Table1.

Cadmium and Lead were analysed in the sample, the concentration of all the heavy metals were below the WHO/FDA permissible limits^20-22. The presence of pesticide residue organochlorine pesticide, organophosphorous pesticides and Pyrethroids were not detected in the sample.

Table 1. Preliminary phytochemical tests for different solvent extract of fruit of Tribulus terrestris Linn.

S. No.	Natural product	Test performed	Result
1.	Alkaloid	Dragendorff’s test	+ve
2.	Coumarin	Alkaline test	+ve
3.	Flavone	Shinoda test	+ve
4.	Steroid	Liebermann-Burchard reagent	+ve
5.	Tannin	Neutral FeCl₃	+ve
6.	Glycoside/Sugar	Molisch’s test	+ve
7.	Terpenoid	Noller’s test	+ve
8.	Saponin	NaOH solution	+ve

Physio-chemical parameters of the fruits of Tribulus terrestries Linn. are tabulated in Table 2. The pH value of 10% w/v aqueous solution is acidic. Deterioration time of the plant material depends upon the amount of water present in plant material. If the water content is high, the plant can be easily deteriorated due to fungus. The loss on drying at 105°C in fruits was found to be 1.52 %. Total ash value of plant material indicated the amount of minerals and earthy materials attached to the plant material. Analytical results showed total ash value content was 11.16 %. The negligible amount of acid-insoluble siliceous matter present in the plant was 1.43 %. The water-soluble extractive value was indicating the presence of sugar, acids and inorganic compounds. The alcohol soluble extractive values indicated the presence of polar constituents like phenols, alkaloids, steroids, glycosides, flavonoids the results given in Table 2.

Table 2: Physico-chemical parameters of fruit of Tribulus terrestries Linn.

S.No.	Parameters	Results
1.	Description	Pale Brown colour
2.	Loss on drying at 105 ⁰C	1.52% w/w
3.	Total Ash	11.43 % w/w
4.	Acid-insoluble ash	1.43 % w/w
5.	Water-soluble extractive	23.00% w/w
6.	Alcohol-soluble extractive	25.50 % w/w
7.	pH (10 % w/v aqueous suspension)	3.10
8.	Bulk density	0.476 gm/ml
9.	Tap density	0.58 gm/ml
10.	Heavy metals
	Mercury	BDL
	Lead	BDL
	Cadmium	BDL
	Arsenic	BDL

*BDL=Below Detectable limit

Thin layer chromatographic technique was used to separate the chemical compounds present in the drug. Various solvent systems were checked to separate the maximum number of chemical compounds in the drug. TLC of the rectified spirit (90%) extract developed in the mobile phase of Toluene: Ethyl acetate (5:1.5 v/v) (Figure. 1). After air drying the plate was not visualized in UV 254 and 366 nm. The plate was then dipped in Vanillin -Sulphuric acid and heated in air oven at 105°C till the spots appeared. After derivatization in visible light various spots at R_fvalue 0.24(grey colour), 0.35 ( grey colour),0.51(grey colour),0.54(bluish grey colour), 0.68(grey colour), 0.78(grey colour), 0. 86 (grey).

TLC of Gokshura (Tribulus terrestris Linn. After derivatization in visible light; Solvent system: Toluene:Ethyl acetate (5:1.5v/v)

CONCLUSION:

Preliminary phytochemical as well as various aspects of the fruits sample were studied and described along with physico-chemical, toxic heavy metal, aflatoxin and TLC studies in authentification adulteration for quality control of raw drugs. The fruits of Tribulus terrestries Linn. exhibits a set of diagnostic characters, which will help to identify the drug in dried condition.

It has been concluded from this study that estimation of heavy metals and pesticides residue and aflatoxin is highly essential for raw drugs or plant parts used for the preparation of compound formulation drugs. The periodic assessment is essential for quality assurance and safer use of herbal drugs.

ACKNOWLEDGEMENT:

The authors are very grateful to Director General, CCRAS, New Delhi and for providing encouragement and facilities for carrying out this work.

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Received on 11.03.2010

Accepted on 15.05.2010

Research J. Science and Tech. 2(2): March –April. 2010: 31-33