PHARMACOGNOSTICAL STANDARDIZATION ON THE ROOTS OF AGNIMANTHA-PREMNA SERRATIFOLIA LINN

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Abstract
Present communication deals with the pharmacognostical standardization on the roots of Agnimantha (Premna serratifolia L. (syn. P. integrifolia L.) belong to the family Verbenaceae. It is an important medicinal plant used in many Ayurvedic formulations. According to Ayurveda the roots and leaves are most useful parts of the plant. It is one among the Dashamula. Macro, microscopical and physicochemical studies were carried out as per the latest Ayurvedic Pharmacopoeia of India, part-1, volume-8 protocols to establish its Pharmacognostical standards. Studies revealed the presence of abundant, simple to compound starch grains with prominent hilum in the centre, cork cells filled with yellowish brown pigmented cells, starch grains in the medullary rays, abundant xylem vessels with bordered pits and tracheids with pits, prism shaped crystals in the cortex region. Phytochemical studies showed the presence of carbohydrates, alkaloids, phenols, tannins etc.
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Reddy et al., IJP, 2018; Vol. 5(7): 426-430. E- ISSN: 2348-3962, P-ISSN: 2394-5583
International Journal of Pharmacognosy 426
IJP (2018), Vol. 5, Issue 7 (Research Article)
Received on 01 March, 2018; received in revised form, 16 April, 2018; accepted, 17 June, 2018; published 01 July, 2018
PHARMACOGNOSTICAL STANDARDIZATION ON THE ROOTS OF AGNIMANTHA -
PREMNA SERRATIFOLIA LINN.
M. Prathapa Reddy *, T. R. Shantha, V. Rama Rao, N. Shiddamallayya and Amit Kumar Dixit
Drug Standardization Research Unit, Regional Ayurveda Research Institute for Metabolic Disorders,
(CCRAS, Ministry of Ayush, Government of India), Ashoka Pillar, Jayanagar, Bangalore - 560011,
Karnataka, India.
ABSTRACT: Present communication deals with the pharmacognostical
standardization on the roots of Agnimantha (Premna serratifolia L. (syn.
P. integrifolia L.) belong to the family Verbenaceae. It is an important
medicinal plant used in many Ayurvedic formulations. According to
Ayurveda the roots and leaves are most useful parts of the plant. It is one
among the Dashamula. Macro, microscopical and physicochemical
studies were carried out as per the latest Ayurvedic Pharmacopoeia of
India, part-1, volume-8 protocols to establish its Pharmacognostical
standards. Studies revealed the presence of abundant, simple to
compound starch grains with prominent hilum in the centre, cork cells
filled with yellowish brown pigmented cells, starch grains in the
medullary rays, abundant xylem vessels with bordered pits and tracheids
with pits, prism shaped crystals in the cortex region. Phytochemical
studies showed the presence of carbohydrates, alkaloids, phenols, tannins
etc.
INTRODUCTION: Agnimantha a sankskrit name
equivalent to Premna serratifolia Linn. (Synonym
P. integrifolia Linn.) is a large, thorny deciduous
shrub or a tree, up to 9 meters in height, common
along the Indian and the Andamans coasts, it also
occurs in the plains of Assam and in Khasi hills. In
Ayurveda the roots and leaves are most used parts
for many ailments. The roots are considered as
astringent, stimulant, liver tonic, laxative,
carminative and antibacterial. Leaves infusion with
pepper is useful in cold and fever, leaf sap is useful
to alleviate headache 1, 2.
QUICK RESPONSE CODE
DOI:
10.13040/IJPSR.0975-8232.IJP.5(7).426-430
Article can be accessed online on:
www.ijpjournal.com
DOI link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.5(7).426-30
Roots are useful in vitiated conditions of vata,
kapha, neuralgia, inflammations, cardiac disorders,
cough, asthma, bronchitis, leprosy, skin disorders,
dyspepsia, flatulence, constipation, fever, diabetes
and anorexia etc.3 In Vietnamese traditional
medicine, the leaves of S. chinensis are used to
treat indigestion, dysuria and dysentery; roots
against indigestion, stomach ache and fever 4. The
root methanolic extract showed better
immunomodulatory activity in experimental mice 5.
A paste of its roots with water and butter fat is
given in Urticaria 6.
Ayurvedic Description of Properties: In terms of
Rsasapanchaka theory of Ayurveda, the properties
are as follows: Rasa (taste) - Tikta (bitter), kaṭu
(pungent), kaṣāya (astringent); Guṇa (quality) -
Rūkṣa (creates dryness), laghu (light for digestion);
Vīrya (potency) - Uṣṇa (Hot in potency); Vipāka
(digestive effect) - Kaṭu (pungent); Karma (action)
Keywords:
Agnimantha, Premna serratifolia,
Premna integrifolia, Pharmacognosy,
Root, Physicochemical
Correspondence to Author:
M. Prathapa Reddy
Research Assistant,
Regional Ayurveda Research
Institute for Metabolic Disorders,
(CCRAS, Ministry of Ayush,
Government of India), Ashoka
Pillar, Jayanagar, Bangalore -
560011, Karnataka, India.
Email: prathapanadri@gmail.com
Reddy et al., IJP, 2018; Vol. 5(7): 426-430. E- ISSN: 2348-3962, P-ISSN: 2394-5583
International Journal of Pharmacognosy 427
- śothahara (Anti inflammatory), vedanāsthāpana
(anodyne or which relieves pain); Doṣaghnatā
(effect on doshas) - Kapha-vātahara (Acts against
the diseases came due to Kapha dosha and Vata
dosha) 7.
As per the 3 volumes of Ayurvedic Formulary of
India, more than 40 formulations has been prepared
by using Agnimantha as one of the ingredient,
some of them are as follows: Amrtarista,
Dantyadyarista, Dasamularista, Agastya Haritaki
Rasayana, Cyavanaprasa, Brahma Rasayana,
Bharngiguda, Dasamula Katutrayakavthacurna,
Dasamulakvathacurna, Dasamulapancakoladikav-
thacurna, Darunagaradikavthacurna, Rasnadikav-
thacurna, Saptasarakavthacurna, Indukanta Ghrta,
Dasamulaghrta, Dasamulasaptalakaghrta, Dadhi-
kaghrta, Dhanvantaraghrta, Mahapancagavyaghrta,
Lasunadighrta, Sukumaraghrta, Dhanvantarataila,
Narayanataila, Prabhanjana Vimardhanataila,
Brha-tmasataila, Mahanarayanataila, Mahavisagar-
bhataila, Sahacaraditaila 8; Brhacchagaladyaghrta,
Guducy-aditaila, Dasamulataila, Madhyama
Narayanataila, Musikadyataila, Vajrakapata rasa,
Vatagankusa rasa, Sirahsuladivajra rasa 9;
Nalikerasava, Dasamularka, Vasisthaharita-
kyavaleha, Caturda-sanga-kvathacurna, Tagaradi-
kvathacurna, Bhunimbadi Astadasangakavthacurna,
Viratarvadiganakavtha-curna etc 10.
Chemical Constituents: Premnacorymboside A,
scutellarioside II, quercetin - 3-rutinoside, leonuriside
A were isolated from the stem bark 4. Alkaloid
contents like premnine, ganikarine, premnazole are
reported in root from different researchers 5. 1β, 3α,
-trihydroxy-pimara-15-ene; 6α, 11, 12, 16-
tetrahy-dxy-7-oxo-abieta-8, 1, 13-triene; 2α, 19-
dihydroxypri -mara-7,15-diene were identified in
root bark 11. It is known as follows in different
Regional languages in India.
Sanskrit: Agnimantha, Ganiparnika, Jjaya,
Vaijayanti, Arani
Bengali: Bhut-bhiravi
Gujarathi: Mothi-arni
Hindi: Agetha
Kannada: Agnimanda, Eegigida
Malayalam: Munna
Tamil: Munnay
Telugu: Pomantai, Peddanarva, Gaebbunelli;
Urdu: Arani 1.
MATERIALS AND METHODS: The roots of
Agnimantha were collected from Budnal village,
Huballi taluk, Dharwad district, Karnataka,
authenticated by Survey of medicinal plant unit,
Regional Ayurveda Research Institute for
Metabolic Disorders, Bangalore. The roots were
shade dried, some are pulverized by mechanical
grinder to get coarse powder and stored in a closed
vessel, to carryout microscopical studies, powder
studies, physicochemical and preliminary phyto-
chemical analysis as per the standard protocols.
Macroscopical, microscopical and powder studies
were carried out as per the standard procedures 12, 13.
Hydro-Alcoholic Extract: 100 g of coarse powder
was taken in an extractor and mixed with 50%
aqueous alcohol, thrice the quantity of raw material
and refluxed for 3 - 4 h. The extract was filtered
through a whatmann no.1 filter paper. The process
was repeated three times and each time the extract
was filtered into the same vessel. The combined
filtrates were concentrated to syrupy consistency
and dried by using a rotary evaporator 14.
Water Extract: 100g of coarse powder was taken
in an extractor and was added with thrice quantity
of distilled water and heated at a temperature 80 oC
for 3 - 4 h. The extract was filtered through a
whatmann no.1 filter paper. The process was
repeated three times and each time the extract was
filtered into the same vessel. The combined filtrates
were concentrated to syrupy consistency and dried
using a rotary evaporator 14.
Thin Layer Chromatography: Shade dried roots
coarse powder was extracted with methanol, water-
alcohol and water by reflux method. TLC studies of
these extracts were carried out by using,
commercially available precoated plates at room
temperature by following standard procedures 14.
Physicochemical Analysis: The powdered drug
was used to carry out various physicochemical
parameters such as total ash, acid-insoluble ash,
loss on drying at 105 OC, water soluble and alcohol
soluble extractive values. The water and water-
alcoholic extracts were dried and subjected to
various tests like total ash, acid-insoluble ash, pH
of aqueous solution and total soluble solids as per
the latest Ayurvedic Pharmacopoeia of India
protocols 14.
Reddy et al., IJP, 2018; Vol. 5(7): 426-430. E- ISSN: 2348-3962, P-ISSN: 2394-5583
International Journal of Pharmacognosy 428
Preliminary Phytochemical Analysis: The
powdered drug was extracted with water, alcohol
and petroleum ether and methanol for carrying out
different preliminary phytochemical tests for
alkaloids, carbohydrates, proteins, phenols, tannins,
saponins, starch, flavonoids and steroids etc. by
using Standard testing protocols 15, 16.
RESULTS:
Macroscopic Characteristics: Roots light brown
or yellowish brown, woody, 6 - 8 cm long and 4 - 5
mm in diameter, brown from outside and yellow
from within, having a short fracture. Roots contain
yellow pigment tannin. Roots are light brown,
woody, branched and cylindrical. Outer surface is
exfoliated at some places and shows longitudinal
striations and wrinkles. Roots possess bland taste
and are slightly aromatic, woody, branched and
tortuous to cylindrical in shape. Surface gets
exfoliated easily and shows prominent longitudinal
striations and wrinkles Fig. 1 and 2.
FIG. 1: LIVE PLANT FIG. 2: DRIED ROOT PIECES
Microscopic Characteristics: The transverse
section shows cork, consisting of 8 - 10 rows of
tangentially elongated and radially arranged
suberised cells. Cork is followed by cortex made up
of 10 - 15 layers of thin walled parenchymatous
cells, closely arranged with little intercellular
spaces. Cortex shows two to three discontinuous
layers of stone cell that are thick walled, lignified
and lodged with prisms of calcium oxalate.
A few cortical cells contain yellowish brown
pigment. Phloem shows thin walled cells and in
between the phloem cells stone cells with prisms of
calcium oxalate crystals are present. The xylem
vessels are of varying size, lignified, found isolated
or in the group of 2 - 3. Medullary rays are 2 - 3
seriate and the cells are pitted and lignified. Starch
is present in the medullar ray cells Fig. 3.
T. S. OF ROOT (DAIGRAMMATIC) T. S. OF ROOT
FIG. 3: MICROSCOPY OF PREMNA SERRATIFOLIA LINN.
Ck: cork, Cam: cambium, ct: cortex, mr: medullary ray, ph: phloem, prc: prismatic crystal of calcium oxalate, stc: stone cell, sg:
starch grains, xyv: xylem vessel
Powder Microscopy: Powdered root is light
brown, with slightly aromatic and bland taste.
Starch is abundant, simple, spherical and cup
shaped. Stone cells are rectangular to oblong and
lodged/ loaded with 3 - 8 prisms of calcium oxalate
which are found scattered also. The vessels and
fibers of the xylem are lignified and found in the
groups of interlocking cells. The xylem vessels
shows bordered pitted thickenings Fig. 4.
Reddy et al., IJP, 2018; Vol. 5(7): 426-430. E- ISSN: 2348-3962, P-ISSN: 2394-5583
International Journal of Pharmacognosy 429
Diagnostic Characters:
Presence of abundant starch grains in the cortex
region.
Presence of prism shaped crystals and stone
cells in the cortex region.
Presence of yellowish brown pigmented cells in
the cork region.
Presence of starch grains in the medullary ray
cells.
Presence of abundant xylem vessels with
bordered pits.
FIG. 4: POWDER STUDY OF PREMNA SERRATIFOLIA L.
Physicochemical Analysis: Physicochemical
parameters such as total ash, acid-insoluble ash,
loss on drying at 105 OC, water soluble and water-
alcohol soluble extractive values were carried out
and results were given in Table 1. The water and
water-alcoholic extracts were dried and subjected
to various tests like total ash, acid-insoluble ash,
pH of aqueous solution and total soluble solids as
per the latest Ayurvedic Pharmacopoeia of India
protocols and results were given in Table 2.
Preliminary Phytochemical Analysis: Preliminary
phytochemical analysis have been carried out for
alkaloids, carbohydrates, proteins, phenols, tannins,
saponins, starch, flavonoids and steroids etc and the
results were given in Table 3.
TABLE 1: PHYSICOCHEMICAL PARAMETERS P.
INTEGRIFOLIA LINN. ROOT POWDER
Name of the parameter
Description
Loss on drying at 105 oC
Total ash
Acid-insoluble ash
Water-soluble extractive
Alcohol-soluble extractive
Water-Alcohol (50:50) soluble
extractive
TABLE 2: PHYSICOCHEMICAL PARAMETERS OF P.
SERRATIFOLIA L. ROOT POWDER EXTRACTS
Name of the
parameter
Values (%) w/w
Hydro-alcoholic
extract
Water
extract
pH (5 % w/v aq. solution)
5.05
4.85
Loss on drying at 105 oC
9.38
10.86
Total ash
5.97
13.28
Acid-insoluble ash
0.25
0.20
Total soluble solids
95.39
92.93
TABLE 3: PRELIMINARY PHYTOCHEMICAL TESTS FOR
DIFFERENT EXTRACTS OF P. SERRATIFOLIA L. ROOT
Natural product
group
Test for natural
products
Presence (+)/
Absence (-)
Alkaloids
Dragendorff’s test
+
Hager’s test
+
Mayers’s test
+
Wagner’s test
+
Carbohydrates
Anthrone test
++
Benedict’s test
++
Fehling’s test
++
Molisch’s test
++
Flavonoids
--
+
Phenols
Ferric chloride test
+
Lead acetate test
+
Proteins
Biuret’s test
+
Millon’s test
+
Saponins
--
+
Steroids
Salkowski reaction
-
Tannins
Ferric chloride test
+
Lead acetate test
+
Thin Layer Chromatography: Thin layer
chromatography was carried out on a precoated
silica gel 60254 plate. Few drops of test solution
applied on a TLC plate as band. Developed the
plate to a distance of 8 cm from the line of
application by using Hexane: Ethyl formate (8:2) as
mobile phase (solvent system).
A B C D E F
FIG. 5: THIN LAYER CHROMATOGRAPHY
A: Powder ext., B: water ext., C: Wat.-Alc. ext. under UV254
nm; D: Powder ext., E: water ext., F: Wat-Alc. ext. after
spraying with 10% H2SO4).
Reddy et al., IJP, 2018; Vol. 5(7): 426-430. E- ISSN: 2348-3962, P-ISSN: 2394-5583
International Journal of Pharmacognosy 430
Dried the plate in air and examined under 254 nm,
recorded the Rf values. Sprayed the TLC plate with
10% sulphuric acid reagent. Heated the plate at
110 OC for about 5 min and recorded the Rf values.
The chromatogram obtained with powder test
solution shows major Rf values under UV-254nm ~
0.06, 0.175, 0.21, 0.30, 0.56 and after spraying at ~
0.06, 0.175, 0.21, 0.30, 0.375, 0.50, 0.56, 0.93. The
hydro-alcoholic extract test solution shows major
Rf values under UV-254 nm at ~ 0.06, 0.175, 0.21
and after spraying at ~ 0.06, 0.175, 0.21, 0.375.
Water extract test solution shows major Rf values
under UV-254 nm at ~ 0.21 and after spraying at ~
0.06, 0.21 Fig. 5.
CONCLUSION: Agnimantha is an important
Ayurvedic drug used in several Ayurvedic
preparations. Studies carried out on the roots of
Agnimantha - Premna integrifolia Linn. helps in
the identification/authentication of different species
of P. integrifolia and also different sources of
Agnimantha. Since agnimantha is also used under
different botanical names like Clerodendrum
phlomidis Linn. f., belonging to the same family
Verbenaceae, this type of studies helps in
differentiating the other sources of agnimantha,
which will be more useful for researchers, students
and Ayurvedic Physicians for identifying the
correct drug and it may also be included in the
Ayurvedic Pharmacopoeia of India.
ACKNOWLEDGEMENT: Authors are thankful
to the Director General, CCRAS, New Delhi for
giving encouragement and facilities to carry out the
work successfully.
CONFLICT OF INTEREST: Nil
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Reddy MP, Shantha TR, Rao VR, Shiddamallayya N and Dixit AK: Pharmacognostical standardization on the roots of agnimantha -
Premna serratifolia Linn. Int J Pharmacognosy 2018; 5(7): 426-30. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.5(7).426-30.
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    • P V Sharma
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    Sharma PV and Vijnana D: Chaukambha Bharati Academy, Varanasi, Edition Reprint, Vol. 2, 2009; 221.
  • The Ayurvedic Formulary of India. Department of Indian Systems of Medicine and Homoeopathy, Ministry of Health and Family Welfare
    • Anonymous
    Anonymous. The Ayurvedic Formulary of India. Department of Indian Systems of Medicine and Homoeopathy, Ministry of Health and Family Welfare, Government of India, Vol. 2, 2000; 1-427.
  • Anonymous: The Ayurvedic Formulary of India. Department of Ayush, Ministry of Health and Family Welfare
    Anonymous: The Ayurvedic Formulary of India. Department of Ayush, Ministry of Health and Family Welfare, Government of India, Vol. 3, 2011; 1-710.
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