NeuroQuantology | DEC 2022 | Volume 20 | Issue 19 | Page 2376-2383| doi: 10.48047/nq.2022.20.19.

NQ99199
Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

Standardization, Extraction and Phytochemical Evaluation of Tribulus

terrestris Fruits
Siva Ganesh Vakkalagadda, Rupesh Kumar M *
Department of Pharmacology, Sri Adichunchanagiri College of Pharmacy, Adichunchanagiri
University, B.G Nagar-571448, Karnataka, India.
Corresponding Author
E-mail: manirupeshkumar@[Link]
Tel. +917348828206.

ABSTRACT: Tribulus terrestris (TT) is a creeping herb which have plenty of medicinal values,
used in Ayurveda and folk medicine in India, China and other countries. The fruits are used to
treat various conditions in traditional Chinese medicine. Tribulus possess benefits to cure
various ailments. The standardization of plants are essential for the identity and quality of
crude drugs for further research. To standardize the plant material, few parameters were
estimated as recommended by World Health Organization. Dried and sieved powder of Tribulus
terrestris fruits were put through organoleptic and microscopic examination followed by
physicochemical evaluation. Later the plant material was defatted, extracted and subjected to
phytochemical screening. The identity and the purity of the plant material was indicated by
macroscopic characteristics and the powdered microscopy is used to determine the specific
internal structures. The quality of powdered raw material by physicochemical methods and
presence of various phytoconstituents by phytochemical analysis reveals the Pharmacognostic
standardization with respect to identity, quality and purity of crude drugs.
KEYWORDS: Tribulus terrestris; Ayurveda; folk medicine; physicochemical; and phytochemical
screening
DOI Number: 10.48047/nq.2022.20.19.NQ99199 NeuroQuantology2022;20(19): 2376-2383
up to one metre high off the ground.
INTRODUCTION
Tribulus is derived from the Grecian word
Tribulus terrestris (TT) is an herb with two "tribolos," indicates "spike fruit." They are
cotyledons that is native to Bharat, western employed to treat various conditions in
countries, South Africa, and Australia. It Chinese folk medicine (CFM), Ayurvedic
belongs to the Zygophyllaceae family, which medication in Bharat, and historical
includes Genus of twenty-five and medicine in Bulgaria [1, 2]. In Telugu,
approximately two hundred and fifty Palleru; In Sanskrit, Gokshur; in English,
species. TT is a creeping plant that highten puncture vine, caltrops; in hindi, it is

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Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

Gokharu; In Gujarathi, Nanagokharu; In Nellore district, Andhra Pradesh. The

Tamil, Nerinjil are some of the common Collected raw material was authenticated
names for TT. It is spread out over a large by Dr. Noorunnisa Begum, Curator,
geographic area. It can be found in Sri foundation for revitalization of local health
Lanka, Kashmir, and many places of India. It traditions, Bangalore-560064. FRLHT Acc.
is the most habitual creeper on road sides No. 5671. After Authentication, the fruits
and open fields particularly in sandy, dry were processed, shade dried for two
and hot regions of India, such as West months and powdered in mixer grinder,
Rajasthan and Gujarat [3]. TT extract packed and stored at room temperature in
contains numerous compounds with diverse air tight container for further study.
biological properties and chemical
structures, including Alkaloids, Carboxylic Chemicals and reagents
acids, Tannins, saponins and terpinoids [4].
The required chemicals and reagents in this
The spirostanol and furostanol saponins like
study were purchased from TCI Chemicals
Hecogenin, Neohecogenin, Tigogenin,
Pvt. Ltd. Kurumandapam, Trichy,
Neotigogenin, Gitogenin, Neogitogenin,
Tamilnadu, India and orbit scientific
Diosgenin, and Sarsapogenin are rich
products, Hyderabad, Telangana, India.
sources of Tribulus. Its extract contains a
steroidal saponin, protodioscin, influence Standardization and extraction
endogenous androgen synthesis by
elevating the release of luteinizing hormone According to WHO guidelines and other
[1, 5]. The whole herb have a variety of Pharmacopoeial specifications the
therapeutic properties, including powdered fruits and extracts were
aphrodisiac, anti-inflammatory, antioxidant standardized [7]. Organoleptic and
and anti-microbial activities. TT is typically microscopic evaluation, physicochemical
helps to treat male-infertility and libido. It parameters like presence of foreign matter,
could be used for immune-modulatory, moisture content, and ash value were
hepatoprotective, hypolipidemic, estimated in dry powder. Fluorescence, and
anthelmintic, and anti-carcinogenic extractive values were measured in various
purposes [6]. solvents [8, 9]. Extraction was performed by
In this article we discuss about the soxhlet apparatus using various solvents
standardization, extraction, and preliminary with change in polarities [10]. Preliminary
phytochemical evaluation of Tribulus phytochemical screening was carried out
terrestris fruits. for all the extracts [11].
MATERIALS AND METHODS Macroscopic and microscopic evaluation
Procurement of Plant material The organoleptic characteristics were noted
The Plant material collected from physically with naked eye compared with
Ulavapadu (Location 15.1667◦N 80.0◦E), the monograph. For the microscopic

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Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

examinations coarse powder is taken and of rudimentary substance these values are
examined under laboratory microscope essential.
[12].
Fluorescence analysis
Foreign matter
A pinch of Tribulus fruit powder was placed
Foreign matter include bacteria, fungal on a glass slide focused under the
molds, animal waste, vehicle pollution, microscope and observed in day light and
insects other than the plant material. The UV light. The powdered sample was placed
presence of foreign matter in the plant in various reagents and observed for
sample were recorded in the form of change in color [15].
percentage weight by weight (% w/w).
Extractive values
Moisture content
The amount of active constituents extracted
The proportions of active ingredients exist with selected menstruums from accurate
in the crude sample were dogged on the quantity of fruit powder determines the
basis of dry weight. Hence the moisture extractive value. Precisely weighed fruit
content must be resoluted. The laid back powder was treated with solvents like n-
technique to determine moisture content is Hexane, n-Butanol, ethanol, methanol, and
to heat the prerequisite amount of drug till water. The extracts were collected,
it gets a persistent weight at 100 ◦C. The subjected to rotavap for solvent recovery
crude compound which go through except water (aqueous) extract [16].
physicochemical change with consequent
loss of weight at 100 ◦C, other methods Phytochemical screening
were employed [13].
Identification of the plant metabolites in
Ash value the extract is one of the preliminary tests
for phytochemical screening. The presence
From the air dried sample, the ash (acid of alkaloids, carbohydrates, flavonoids, fats,
insoluble ash, total ash, and water soluble glycosides, proteins, saponins, sterols, and
ash) values were resoluted by the method tannins [17-20].
described in Indian pharmacopoeia [14].
Ash value is used to determine the ratio of RESULTS
inorganic materials (calcium, carbonates, Macroscopic and microscopic evaluation
magnesium, phosphates, potassium Tribulus terrestris is a silky hirsute shrub
silicates and sodium) exist in the crude with opposite leaves with equal number of
compound. To govern the quality and purity leaflets on both sides of midline with yellow
flowers having characteristic odour. The

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Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

height of plant is approx. 70 cm when fruits and its powder were represented in
measured. The fruits are small, nearly figure 1. The powdered microscopy
round shaped with 2-5 sharp spikes consists contains numerous epidermal trichomes
of seeds within them. Seeds are wet and parenchyma cells, polygonal thick-walled
oily in nature. The spiky fruits appears as cells, epidermal cells, sclerenchymatos cells,
cow hoof therefore it is also called as fibre sclereids, and few crystals. The
gokshura [3]. Roots appears as slender with structures of few cells were depicted in
bunch of rootlets with cylindrical shape. The figure 2.
macroscopic characteristics of plant, dried

Figure 1: Macroscopy of Tribulus terrestris herb. A: Whole plant of Tribulus terrestris. B.

Dried fruits of Tribulus terrestris. C. Tribulus terrestris fruits powder.

Figure 2: Powdered microscopy of Tribulus terrestris fruits. A: Fibre tracheids and Brachy
sclereids. B. Calcium oxalate crystals. C. Trichomes.
Physicochemical parameters
Physicochemical parameters like foreign matter, moisture content, and ash values (Total ash,
Acid insoluble ash, and Water soluble ash) were indicated in table 1.
Table 1: Foreign matter, Moisture content (loss on drying), ash values (Total ash, Acid insoluble
ash, and Water soluble ash).
Physicochemical parameters (% w/w)
Foreign matter 0.26

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Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

Loss on drying 3.84

Total ash 7.36
Acid insoluble ash 1.2
Water soluble ash 4.1

Fluorescence analysis
The change in the color of powdered fruit sample with various solvents were reported in table
2.
Table 2: The change in color under day light and UV light.
TREATMENT Day light Tribulus terrestris fruit powder under UV
Short UV (264 nm) Long UV (366 nm)
Fruit Powder Pale Brown Dark Green Pale Green
as such
Fruit Powder + Dark Green Pale Green Greenish Brown
1N sodium
hydroxide
Fruit Powder + Greenish Black Pale Black
1N sulphuric acid Brown
Fruit Powder + Dark Brown Pale Green Light Green
1N hydrochloric acid
Fruit Powder + Yellow Dark Green Black
1N nitric acid Brown
Fruit Powder + Pale Brown Dark Brown Greenish Brown
acetic acid
Fruit Powder + Dark Green Pale Green Purplish Green
5% iodine

Extractive values
The extractive values of each extract in individual solvent were expressed in table 3.

Table 3: Extractive values of Tribulus terrestris fruit powder in various solvents.

S. No Solvent (% w/w)
1 n-Hexane 28.66
2 n-Butanol 15.68

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Siva Ganesh Vakkalagadda / Standardization, Extraction and Phytochemical Evaluation of Tribulus terrestris Fruits

3 Ethanol 9.56
4 Methanol 8.52
5 Water 7.1

Preliminary phytochemical screening

The presence of various phytoconstituents were tabulated in table 4.

Table 4: Phytochemical screening Tribulus terrestris fruits.

Tests Methanolic ethanolic extract Aqueous
extract extract
Alkaloids + ++ ++
Carbohydrates ++ ++ +
Saponins ++ ++ -
Steroids ++ ++ +
Tannins + ++ -
(++) Highly present (+) present (-) absent
DISCUSSION precise identity and purity of a basic
The macroscopic and microscopic material. This can be done by evaluating its
characteristics reveals the identity and organoleptic, microscopic, and other
purity of the plant material. where as the parameters. The concurrent study provides
physico-chemical properties are indicted in many qualitative and quantitative principles
(% w/w). The presence of foreign matter is of Tribulus terrestris fruits. This
0.26, loss on drying is 3.84, total ash 7.36, standardization will provide referential data
acid insoluble ash 1.2 and water soluble ash on the identity, quality, and purity of the
4.1. The fluorescence for powder was crude drugs, preventing them from being
observed as such, under short and long UV. degraded by a similar substandard product.
The extractive values of n-Hexane is 28.66 , Acknowledgements
for n-Butanol 15.68, Ethanol 9.56, Methanol This research was supported by Sri
8.52 and Water 7.1 and various Adichunchanagiri College of Pharmacy,
phytochemicals like presence of alkaloids, Adichunchanagiri University.
carbohydrates, saponins, steroids and Conflicts of Interest
tannins were confirmed. The authors declare no conflict of interest.
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