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A Comparative Study of the Quality Assessment of Vitex negundo Linn Leaves

Collected from Three Different Geographical Locations

Article · January 2011

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 Asian J. Research Chem. 4(6): June, 2011

ISSN 0974-4169 www.ajrconline.org

RESEARCH ARTICLE

A Comparative Study of the Quality Assessment of Vitex negundo Linn

Leaves Collected from Three Different Geographical Locations
Ajay Kumar Meena1*, Uttam Singh2, Arjun Singh3, Sudeep Mishra3, M. M. Rao1, M. M. Padhi3,
Ramesh Babu3 and A. Gaurav2
1
National Institute of Ayurvedic Pharmaceutical Research, Patiala- 147001, Punjab, (India)
2
School of Pharmaceutical Sciences, Shobhit University, Meerut, UP,(India)
3
Central Council for Research in Ayurveda and Siddha (CCRAS), Janakpuri, Delhi-110058.
*Corresponding Author E-mail: ajaysheera@gmail.com

ABSTRACT:
Vitex negundo Linn (Five-leaved chaste tree), Hindi-Nirgundi, Sanskrit-Sindhuvara, Tamil and Malyalam-Vennochi,
Telugu-Nalla vavili and Punjabi-Marwan (Family: Verbenaceae), is an important medicinal plant found throughout
India. All parts of this plant especially the leaf and root extracts are being used in various formulations in Ayurveda
and Unani systems of medicine. Vitex negundo Linn is pungent, bitter and astringent in taste and according to
Ayurveda , it alleviates vata and kapha doshas, but aggravates pitta dosha. It possesses light and dry attributes. Its
leaves and seeds are widely used externally for rheumatism and inflammations of joints and also reported to have
insecticidal properties. Internally, decoction of its leaves is taken as diuretic, expectorant, vermifuge, tonic and
febrifuge. The chemical components of the essential oil of leaf isolated from the plant are used to treat colds and
coughing spells. Vitex negundo Linn is used in Classical Ayurvedic Preparations like Nirgundi kalka, Nirgundi ghrta,
Nirgundi kvatha, Vranasodhana taila, Visagarbha taila etc. The evaluation studies of Pharmacopoeial standards,
phytochemical parameters suggested that the observed physiochemical parameters are of great value in quality control
and formulation development of Vitex negundo Linn. Physicochemical parameters like preliminary characteristics,
toxic heavy metals, and aflatoxin analysis were carried out. The study revealed that the results on different parameters
of the crude drug will be useful in identification and control of adulterations.

KEYWORDS: Ayurveda, Vitex negundo Linn, Toxic metals, Aflatoxin and Quality control.

INTRODUCTION:
Vitex negundo Linn (Five-leaved chaste tree), Hindi-
Nirgundi, Sanskrit-Sindhuvara, Tamil and Malyalam-
Vennochi, Telugu-Nalla vavili and Punjabi-Marwan
(Family: Verbenaceae), is an important medicinal plant
found throughout India1,2. Grows gregariously in wastelands
and is also planted as a hedge-plant. It is an erect, 2–5 m in
height, slender tree with quadrangular branchlets distributed
throughout India. The leaves have five leaflets in a
palmately arrangement, which are lanceolate, 4–10 cm long,
hairy beneath and pointed at both ends. The bluish purple
flowers are numerous. The fruit is succulent, black when
ripe, rounded and about 4 mm in diameter.3,4
Received on 22.02.2011 Modified on 07.03.2011
Accepted on 12.04.2011 © AJRC All right reserved
Asian J. Research Chem. 4(6): June, 2011; Page 986-989
Vitex negundo Linn is pungent, bitter and astringent in taste
and according to Ayurveda , it alleviates vata and kapha
doshas, but aggravates pitta dosha. It possesses light and
dry attributes. It has antipyretic, anti-arthritic and anti-
inflammatory properties and is used in diseases like fever,
worms, dermatoses, adenitis and splenic diseases etc5
Phytochemical studies on Vitex negundo Linn have afforded
several types of compounds, such as volatile oils6-9,
lignans10,11, Flavonoids 12-14, iridoids15-17, terpenes
(triterpenes, diterpenes, sesquiterpenes)18-19, and steroids.20
The most Flavanoid, glycoside from Leaves of Vitex
negundo Linn of ethanolic extract is 5-hydroxy-3, 6,7-
trimethoxy-2-(3,4-dimtoxypheny)-4H-chrome-4-on and 5,
7-dihydroxy-2- (3,4- dihydroxyphenyl) -4H-chromen-4-
one.21 Methanolic extract also contains, Negundoside,
Agnuside, Vitegnoside22. Bark of Vitex negundo Linn. p-
Hydroxybenzoic acid and -sitosterol were isolated, and
identified from the methanol and hexane extracts of Vitex
negundo.

986
 Asian J. Research Chem. 4(6): June, 2011
From the seed of Vitex negundo Linn the following
compound Isolated are -acetoacetate fraction, two
phenylnaphtha- lene-typelignans and identified as 6-
hydroxy-4-(4-hydroxy-3-methoxy-phenyl)-3-hydroxy-
methyl-7-methoxy-3,4-dihydro-2-naphthaldehyde and
vitedoamine A, both of which have been previously
reported and isolated from the seeds of Vitex negundo.
Powdered roots are used for piles and as a demulcent for
dysentery. It also used in dyspepsia, colic, rheumatism,
worms, boils and leprosy.23 The roots are used as an
antidote to snake venom which contain a physiochemical evaluation. Powder of the samples was
furanoeremophilane, Tyrosinase inhibitory lignin’s from the
methanol extract.24
Hepatoprotective activity of Vitex negundo Linn was
investigated against hepatotoxicity produced by
administering a combination of three anti-tubercular drugs
isoniazid -7.5 mg/kg, rifampin-10 mg/kg and pyrazinamide-
35 mg/kg from the ethanolic extract of leaves. Vitex
negundo Linn contains many polyphenolic compounds,
terpenoids, glycosidic iridoids and alkaloids. Since
polyphenolic compounds have high antioxidant potential,
the antioxidant potency of Vitex negundo was investigated
by employing various established in vitro systems, such as
2, 20-azino-bis 3-ethyl benzothiazoline- 6-sulfuric acid
/Lipid Peroxide /Superoxide/Hydroxyl radical scavenging
and iron ion chelation. The anti-inflammatory, analgesic
and antihistamine properties of mature fresh leaves of Vitex
negundo Linn claimed in the Ayurvedic medicine were
established in Rats by orally treating water extract of the
leaves to rats. The carrageenan-induced rat paw oedema
was significantly suppressed in inversely dose-dependent
manner. anti-hyperglycemic activities.25 Vitex negundo
exhibited significant activity against E. coli, K. aerogenes,
P. vulgaris and P. aerogenes at all dosages. Extract of leaves
of V. negundo showed activity against bacteria.26 The
antiasthmatic activity was evaluated by various
experimental models like mast cell degranulation. 27 Anti-
implantation activity was found in the methanolic extract of
leaves of Vitex negundo Linn, 28 Xanthine oxidase
inhibitory activity was assayed from Vitex negundo.
The methanolic root extracts of Vitex negundo Linn. and
Emblica officinalis Gaertn. were explored for the first time
for anti snake venom activity. The plant (Vitex negundo and
Emblica officinalis) extracts significantly antagonized the
Vipera russellii and Naja kaouthia venom induced lethal
activity both in vitro and in vivo studies.29 Petroleum ether
extracts of the leaves of Vitex negundo were evaluated for
larvicidal activity against larval stages of Culex
tritaeniorhynchus in the laboratory.30 Crude aqueous extract
of Vitex negundo Linn leaves are investigated for laxative
activity.31 Ethanolic extracts of Vitex negundo were taken
for anathematic activity against Indian earthworm Pheritima
posthuma.
987
MATERIALS AND METHODS:
Plant material:
The Leaves of Vitex negundo Linn Were collected in
September, 2009 from Gwalior M.P. (Sample S1), National
Institute of Ayurvedic Pharmaceutical research, Patiala,
Punjab (Sample S2) and Chennai (Sample S3) specimen
were identified and authenticated at the National Institute of
Ayurvedic Pharmaceutical Research (NIAPR), Patiala
(Punjab). Leaves were washed in running water and air-
dried. The fresh leaves were then studied for
used for chemical analysis. Physicochemical studies like
total ash, water soluble ash, and loss on drying at 105°C,
TLC and extractive values were carried out as per the
WHO/AOAC guidelines.32,33
RESULTS AND DISCUSSIONS:
Preliminary phytochemical study:
Preliminary phytochemical results showed the presence or
absence of certain phytochemical constituents in the Vitex
negundo Linn sample. Hexane extract showed the presence
of Fixed oils and fats, alcohol extracts gave positive results
for Carbohydrate, Alkaloid, Flavanoid, steroid, tannins,
Phenolic compounds, Saponins and gave negative results
for Anthraquinones, Fixed oils and fats. Water extract gave
positive results for Alkaloid, Flavanoid, steroid, tannins,
Phenolic compounds and gave negative results for
Carbohydrate, Saponins, Fixed oils and fats shown in Table
1.
Physico-chemical parameters of the leaves of Vitex negundo
Linn are tabulated in Table 2. 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
easily be deteriorated due to fungus. The loss on drying at
105 °C in samples S1, S2 and S3 were found to be 9.47, 8.5
and 8.0 % w/w respectively. 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 samples S1, S2 and S3 content were
8.0, 6.4 and 8.2 % w/w respectively. The amount of acid
insoluble siliceous matter present in the plant samples were
0.31, 0.81 and 0.87 % w/w respectively. The water-soluble
extractive value was indicating the presence of sugar, acids
and inorganic compounds. The water soluble extractive
value in the drug sample were 28.0 (S1), 28.34(S2) and
28.76(S3) % w/w and alcohol soluble extractive value were
16.44 (S1), 17.19 (S2) and 16.87 (S3) % w/w. The alcohol
soluble extractive values indicated the presence of polar
constituents like phenols, alkaloids, steroids, glycosides,
flavonoids and secondary metabolites present in the plant
sample. Other parameters, like pH, are given in Table 2.
Aflatoxin B1, B2, G1, G2 is also not detected in the drug
sample Heavy metals and microbial contaminations are
below permissible limit in the samples.
 Asian J. Research Chem. 4(6): June, 2011

Table 1. Preliminary phytochemical tests of various solvent extract of Vitex negundo Linn.
S. No. Phytoconstitunts Successive extraction of Sample, S1, S2, S3
n-Hexane Alcohol Water
1. Carbohydrate -ve +ve -ve
2. Alkaloid -ve +ve +ve
3. Flavanoid -ve +ve +ve
4. Steroid -ve +ve +ve
5. Tannin -ve +ve +ve
6. Phenolic compounds -ve +ve +ve
7. Saponins -ve +ve -ve
8. Anthraquinones -ve -ve +ve
9. Fixed oils and fats +ve -ve -ve

Table 2. Analysis of physico chemical parameters of Vitex negundo Linn

Parameters S1 ( Gwalior) S2 (Patiala) S3 ( Chennai)
pH (10% w/v aqueous solution) 4.8 5.0 4.8
Total Ash (%w/w) 8.0 6.4 8.2
Acid insoluble ash (%w/w) 0.31 0.81 0.87
Water soluble extract (%w/w) 28.0 28.34 28.76
Alcohol soluble extract (%w/w) 16.44 17.19 16.87
Loss on drying at 105°C (%w/w) 9.47 8.5 8.0

2gm of sample was soaked overnight in 20 ml of 90%
ethanol. The solutions were continuously stirred for 6 hrs
and kept for next 18 hrs., next day filtered, dried and made
10% solution. The solution was applied on Merck
Aluminium pre-coated plate with silica gel 60 F254 of 0.2
mm thickness. The plate was developed in Toluene: Ethyl
acetate (9: 2 v/v). After air dry the plates were visualized in
UV 254 and 366 nm. The plates were dried and then
dipped in Vanillin- Sulphuric acid and heated at 105 ° C till
the spots appeared.
TLC of the ethanol extract developed in the mobile phase of
Toluene: Ethyl acetate (9: 2 v/v) and observed under UV
254 nm 3 spots at Rf value 0.24,0.35 (Pale yellow) and 0.47
(Brown); under UV 366 nm showed 3 spots at Rf 0.24, 0.35
and 0.47 (Pale red), and after derivatization with Vanillin-
Sulphuric acid, showed 5 spots at Rf 0.37 (Pale green), 0.43
(Pale blue), 0.50 (Pale green), 0.62 (Blue) and 0.68 (Pale
blue). Heavy metals contamination is below permissible
limit in the samples and Aflatoxin B1, B2, G1, G2 are also not
detected in the samples.
CONCLUSION:
Physiochemical parameters that are loss on drying at 105ºC,
Total ash, Acid insoluble ash and extractive values on
comparison revealed that extractive value is highest in
sample S3. Highest extractive values show the highest
active component in sample S2, S3 comparison with sample
S1. It was observed that there is variation in the values
obtained on analysis of the various samples of Vitex
negundo Linn collected from different regions. It has been
concluded from this study that estimation of physico-
chemical parameters, heavy metals and aflatoxin is highly
essential for raw drugs or plant parts used for the
preparation of compound formulation. The periodic
assessment is essential for quality assurance and safer use
of herbal drugs, So that study can be considered as the
Pharmacopoeia standard and helpful to confirm the
988
genuineness of the single plant drug to be used for the
preparation of effective Ayurvedic formulation.
ACKNOWLEDGEMENT:
The authors wish to thank the Director, Shobhit University
Meerut for providing encouragement to complete the work
successfully.
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