pharmacognostical and preliminary phytochemical investigations

Journal of Herbal Medicine and Toxicology 4 (2) 113-118 (2010)
ISSN : 0973-4643 Original Article
PHARMACOGNOSTICAL AND PRELIMINARY PHYTOCHEMICAL
INVESTIGATIONS ON VERBESINA ENCELIOIDES BENTH. ROOTS
Rakesh K. Sindhu*, Neeru Vasudeva and S.K. Sharma
Division of Phytochemistry and Pharmacognosy, Department of Pharmaceutical Sciences, Guru
Jambheshwar University of sci. &Technology, Hisar, 125001, India.
*Corresponding author email: E-mail: rakeshsindhu16@gmail.com
Received 4th Feb, 2010, Revised- 13th April, 2010, Accepted- 2nd May,2010
ABSTRACT: Phytochemical studies of the Verbesina encelioides Benth. Roots were
carried out in present study. Morphology of the plant, microscopy of the root. Total
ash, acid insoluble ash, water insoluble ash and sulphated ash were 7.86%, 5.45%,
4.32%, 7.80% respectively. The extractive values i.e. petroleum ether, chloroform,
ethyl acetate, ethanol and aqueous extract were 5.7%, 10%, 5.5%, 4.5%, 10.5%. The
alcoholic and aqueous extracts were screened for presence of amino acid and
carbohydrates. The extracts showed the presense of two amino acid viz. nor-lucine
and ornithine and two carbohydrates, D-ribose and sucrose. The preliminary
phytochemical screening of alcoholic and aqueous extracts was performed. The
presence of flavonoids, carbohydrates, saponins, phenolic compounds, and sterol in
alcoholic extract was observed. Flavonoids, carbohydrates, saponins, amino acids
and phenolic compounds were present in aqueous extract.
Key Words: Verbesina encelioides Benth., pharmacognostical, phytochemical screening,
INTRODUCTION
Pharmacognostical study is the preliminary step in
the standardization of crude drugs. The detailed
pharmacognostical evaluation gives valuable
information regarding the morphology, microscopical
and physical characteristics of the crude drugs.
Pharmacognostic studies have been done on many
important drugs, and the resulting observations have
been incorporated in various pharmacopoeias [1].
There are a number of crude drugs where the plant
source has not yet been scientifically identified. Hence
pharmacognostic study gives the scientific information
regarding the purity and quality of the plant drugs [2].
Verbesina is a small genus native to the Southwestern
United States and Maxican platueau, the species of
which occur primerly in the arid grass and scrublands.
Some species have ornamental and economical value.
Verbesina encelioides (Astracae) is weedy and has
extentedsed into several regions of the world. Other
species are i.e. (Verbesina boliviana, Verbesina
cinera, Verbesina eggersi, Verbesina gignonta,
Verbesina macrophylla etc.) Verbesina encelioides
[(Cav.) Benthum & Hooker fil. Gray], Synonym:
Ximenesia encelioides, Common name: Crown beard
[3, 4]. The plant is used traditionally to treat stomach
diseases, heamorrhoides. The roots are used for
retention of water, bladder inflammation and also as
a blood purifier [5]. Leaves are used as a poultice to
relieve sore legs to treat rheumatism and the juice is
used as a laxative [6]. The major phytoconstituents
from the different parts are terpenoids, flavonoids,
aromatic compounds etc.[7]. The present study was
for identificattion of plant and explored for the
phytochemical and screening of biological active
compounds.
MATERIALS
The plant of Vebesina encelioides Benth. root was
collected during the month of the July 2007 from Village
Dinod, Bhiwani (Haryana), North India.
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Journal of Herbal Medicine & Toxicology
Table 1 : Behaviour of root powders with different chemical reagents
Sr. No Treatment Color of powder
1 Powder as such Light brown
2 Powder + 1 N HCl Dark brown
3 Powder + 1N NaOH No change
4 Powder + Acetic Acid Light pinkish
5 Powder + 5% Ferric chloride Creamish
6 Powder + Picric acid Greenish yellow
7 Powder + HNO3 + Ammonia solution Light brown
8 Powder + 5% Iodine Dark brown
9 Powder + 1N HNO3 Blackish green
Table 2 : Fluorescence nature of root powder under ultra violet (UV) and visible radiations
Sr.
no. Treatment
114
Long UV
(365 nm)
Short UV
(254 nm)
Visible
1 Powder as such Dark brown Light brown Light brown
2 Powder + 1N HCl Light brown Light brown Light brown
3 Powder + 1N NaOH Light brown Light yellow Light brown
4 Powder + 50% HNO3 Light green Light brown Light brown
5 Powder + Acetic acid Light brown Light yellow Light brown
6 Powder + Picric acid Light brown Dark brown Light brown
7 Powder + 1N NaOH in methanol Light greenish Dark brown Light brown
8 Powder + FeCl3 Creamish Light brown Light brown
9
Powder + 1N NaOH in methanol + Nitrocellulose in
amyl acetate
Light yellowish Light greenish Light brown
10 Powder + 1N HCl + Nitrocellulose in amyl acetate Light greenish Brown Light brown
11
Powder + 1N NaOH + Nitrocellulose in amyl
acetate
Table 3 : Determination of Ash Values of Verbesina encelioides Benth
Yellowish green Light green Light brown
S. No. Ash type Percentage of Ash
1. Total ash 7.8% w/w
2. Acid insoluble ash 5.32% w/w
3. Water soluble ash 3.94% w/w

Table 4 : Extractive values of root powder with various solvents
115
Sindhu et al.
Sr. No. Solvent Extraction period (h) Colour of extract Extractive value (%) w/w
1. Petroleum ether (60-80°C) 24 Light brown 5.7
2. Chloroform 24 Light brown 10
3. Ethyl acetate 24 Yellowish brown 5.5
4. Ethanol 24 Brown 4.5
5. Aqueous 24 Dark brown 10.5
Table 5 : Amino acids analyzed by paper chromatography
Sr. No. Amino Acids Rf Value Amino acids detected
1. Alanine 0.32 –
2. 2-Aminobutyric acid 0.70 –
3. Arginine 0.15 –
4. Aspartic acid 0.16 –
5. Glutamic Acid 0.18 –
6. Glycine 0.16 –
7. Histidine 0.14 –
8. Lysine 0.18 –
9. Methionine 0.37 –
10. Norleucine 0.10 +
11. Ornithine 0.10 +
12. Phenylalanine 0.25 –
13. Tyrosine 0.47 –
+ve : Detected; –ve : Not detected Solvent system : n-butanol : water: glacial actic acid (4:1:1, upper phase)
Spray reagent : Ninhydrin
Table 6 : Carbohydrates analyzed by paper chromatography
Sr. No. Carbohydrates Rf Value Carbohydrate Detected
1. Galctose 0.646
2. Maltose 0.60 -
3. Lactose 0.622 -
4. D-Ribose 0.525 +
5. Sucrose 0.629 +
6. Fructose 0.666 -
7. D-Xylose 0.711 -
8. L-Arabinose 0.666 -
+ve : Detected; –ve : absent Solvent system : n-butanol : glacial acetic acid : water (2:1:1)
Spray reagent : Anisaldehyde in sulphuric acid

Journal of Herbal Medicine & Toxicology
Table 7 : Prelimenry phytochemical screening
Sr. no. Plant Constituents Test / Reagent Ethanol extract Aqueous extract
1. ALKALOIDS
Mayer’s reagent
Dragendroff’s reagent
Wagner’s reagent
2. GLYCOSIDES
Killer-Killani test
Sodium nitropruside test
Borntrager test
3. CARBOHYDRATES
Molisch’s reagent
Fehling solution
4. STEROLS
Liebermann-Burchard’s test
Salkowski test
Hesses reaction
Hersch reaction
5. SAPONINS
Foam test
Sodium bicarbonate test
6. PHENOLIC COMPOUNDS & TANNINS
Ferric chloride solution
Lead acetate solution
7. PROTEINS & AMINO ACIDS
Biuret test
Millon’s reagent
Ninhydrin reagent
8. FLAVANOIDS
Shinoda/Pew test
Ammonia test
+ve: Present; -ve: Absent
The plant material was taxonomically identified and
authenticated by Dr. H.B. Singh, Head, Raw materials
Herbarium and Museum division, with
ref.no.NISCAIR/RHMD/Consult/2007-08/890/74.
The voucher specimen has been deposited in the
116
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herbarium section of the Pharmacognosy Division,
Department of Pharmaceutical Sciences, Guru
Jambheshwar University of Science and Technology,
Hisar for further reference. The roots were dried
under shade, sliced into small pieces, pulverised using
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a mechanical grinder and stored in an air tight
container for further use.
METHODS
The crude drug was evaluated for organoleptic
properties shape, size. colour, odour, taste, fracture
and texture were noted as per Wallis [8]. Quantitative
microscopy was determined by methods prescribed
by Trease and Evans [9].
The ash values, extractive values with various reagents
and were determined as per the WHO guideline [10].
The behaviour of powdered roots with various
chemical reagents was studied [11]. The fluoroscence
characters of the powder with various acids were
observed under visible light and UV light as per the
procedure [12]. Extractive values were performed
with various solvents like petroleum ether, chloroform,
ethyl acetate, alcohol and water was performed as
per Indian Pharmacopoeia [14]. Amino acids analyzed
by paper chromatography. Preliminary phytochemical
tests of the extracts were performed using specific
reagents through standard procedures [15] .
RESULTS AND DISCUSSION
The pharmacognostical studies i.e. morphological
study of the root of the plant was performed. The
root is a typical tap root. The fresh roots were light
yellowish color which turned light brown on drying.
The bark of the root was thin. The roots are long,
thick, hard and cylindrical about 1 mm to 1.2 cm in
diameter having smooth surface with secondary
rootlets. The root breaks with short fracture when
dry. It has characteristic odor and in transverse section
it shows the features of a dicoat root (Figure 1). The
microscopy of root revealed the presence of Cortex,
Cutical, Epidermis, Primary phloem, Xylem
parenchyma, Xylem vessels (Figure 2). Behaviour
of root powders with different chemical reagents
(Table 1). Fluorescence nature of root powder under
ultra violet (UV) and visible radiations (Table 2). Total
ash, acid insoluble ash, water insoluble ash and
sulphated ash were 7.86%, 5.45%, 4.32%, 7.80%
respectively (Table 3). The extractive values i.e.
petroleum ether, chloroform, ethyl acetate, ethanol and
aqueous extract were 5.7%, 10%, 5.5%, 4.5%, 10.5%
(Table 4).. The alcoholic and aqueous extracts were
screened for presence of amino acid (Table 5) and
carbohydrates (Table 6). The extracts showed the
presense of two amino acid viz. nor-lucine and
ornithine and two carbohydrates, D-ribose and
sucrose. The preliminary phytochemical screening of
alcoholic and aqueous extracts was performed. The
presence of flavonoids, carbohydrates, saponins,
phenolic compounds, and sterol in alcoholic extract
117
Sindhu et al.

Journal of Herbal Medicine & Toxicology
was observed. Flavonoids, carbohydrates, saponins,
amino acids and phenolic compounds were present
in aqueous extract (Table 7). This is first ever
pharmacognostical study carried out on the roots of
Verbesina encelioides Benth.
REFERENCES
[1]. Sharma SK (2004). Recent approach to herbal
formulation development and standardization, http/
/pharmainfo.net.
[2]. Dhanabal SP, Suresh B, Sheeja E and Edwin E.
Pharmacognostical studies on Passiflora
quadrangularis. Indian Journal of Natural
Product,s (2005) 21(1): 9-11.
[3]. Ball WS, Crafts AS, Robbins. Principal of weeds.
Weed of califonia, California agricultural extension
service. California. (1951) 13-15.
[4]. Parker KF. An illustrated guide to Arizona
weeds.university of Arizona press. Arizona (1972)
322-323.
[5]. Bhattacharjee SK. Handbook of medicinal plants,
pointer publication Jaipur, first edn. (1998) 371.
[6]. Parrota JA. Healing plants of penisular India CABI
publishinghouse, USA, (2001), 155-156.
“ERRATUM”
118
[7]. Bohlmann F, Grenz M, Gupta RK, Dhar AK, Ahmed
M, King RM and Robinson H [1980]. Eudesmane
derivative from Verbesina species. Phytochemistry,
19 : 2391-97
[8]. Wallis TE [1989]. Text Book of Pharmacognosy (CBS
publishers and Distributors, Delhi 356 – 549.
[9]. Evans WC [1996]. Trease and Evans.
Pharmacognosy, WB Sacenders Company Ltd.
London, 14 th Edn. pp. 194.
[10]. WHO [1998]. Quality Control methods for medicinal
plants material. World Health Organization, Geneva,
AIT US publisher and distributor Delhi.
[11]. Brain KR and Turner TD. The practical evaluation of
phytopharmaceuticals, Wright-Scientechnia, (1975)
6, 81.
[12]. Kokoshi J, Kokoski R and Slama FJ. Fluorescence
analysis of powered vegetable drugs under
ultraviolet radiation , J Am Pharm Assoc, (1958) 47,
75-77.
[13]. Sharma SK and Ali M. Amino acid and carbohydrate
composition of stem bark of some cultivars of
Mangifera indica (Mango). Journal of Indian
Chemical Society, (1992) 69, 891-892.
[14]. Peach and Tracey MV. Modern Methods of Plant
Analysis, Vol. III (Springeand Verlag, Berlin) (1955)
321-322.
The Editorial board of JHMT regrets the publishing of manuscript entitled
“Neuroprotective effects of Asparagus racemosus Linn. root extract : An
experimental and clinical study”; as it has been published elsewhere. This mistake
had been due to over sight.