12 Chapter2

REVIEW OF LITERATURE
Chapter 2
2.
INTRODUCTION
2.1. GENUS PREMNA: AN OVERVIEW

2.1.1. TAXONOMIC HISTORY OF THE GENUS PREMNA
2.1.1.1. INDIAN HISTORY OF THE GENUS PREMNA
2.1.1.2. PERPLEXITY IN CLASSIFICATION OF THE GENUS PREMNA
2.1.2. DISTRIBUTION OF THE GENUS PREMNA
2.2. PREMNA SERRATIFOLIA: AN OVERVIEW
2.2.1. TAXONOMIC HISTORY OF PREMNA SERRATIFOLIA
2.2.2. DISTRIBUTION OF PREMNA SERRATIFOLIA
2.2.3. POLYMORPHIC STATUS OF PREMNA SERRATIFOLIA
2.3. PREMNA SERRATIFOLIA: A POTENTIAL MEDICINAL PLANT
2.3.1. TRADITIONAL MEDICINAL USES OF PREMNA SERRATIFOLIA
2.3.2. PREMNA SERRATIFOLIA IN AYURVEDIC FORMULATIONS
2.3.3. DESCRIPTION OF PREMNA SERRATIFOLIA UNDER SANSKRIT NAMES IN
TRADITIONAL MANUSCRIPTS
2.3.4. CONTROVERSIAL STATUS OF PREMNA SERRATIFOLIA IN AYURVEDA
2.4. PHARMACOGNOSTIC, PHYTOCHEMICAL AND PHARMACOLOGICAL
STUDIES ON PREMNA SERRATIFOLIA
2.4.1. PHARMACOGNOSTIC STUDIES ON PREMNA SERRATIFOLIA
2.4.2. PHYTOCHEMICAL STUDIES ON PREMNA SERRATIFOLIA
2.4.3. PHARMACOLOGICAL STUDIES ON PREMNA SERRATIFOLIA
2.5. CONCLUSION
Chapter 2
2. Introduction
The taxonomic status of Premna serratifolia L. has been a topic of heated discussion
among taxonomists from the early period of Linnaeus due to extremely polymorphic
features exhibited by this species reported from different geographic regions of the
world. In the Indian traditional Ayurvedic system of medicine, the plant Agnimantha
(Sanskrit name) is described as a highly valuable ingredient of Dashamoolam. Two
different species viz., Premna serratifolia L. and Clerodendrum phlomidis L.f.
(Clerodendron phlomidis) are equated to the source plant Agnimantha, providing a
controversial drug status to these two medicinal plants. Consequently, for the
preparation of Ayurvedic formulations having Agnimantha as ingredient, Clerodendrum
phlomidis is used as the source drug in North India and Premna serratifolia is used in
South Indian States. However, in Kerala, various morphotypes of Premna serratifolia
are used as the source drug and Clerodendrum species viz., Clerodendrum inerme is
used as a substitute drug of Clerodendrum phlomidis. Hence, an attempt is made in the
present investigation to resolve the ambiguity regarding the taxonomic as well as
Ayurvedic status of Premna serratifolia using the most reliable and sophisticated tools
and techniques in pharmacognosy and phytochemistry. In the above context, a brief
review of the available literature on the genus Premna and the species Premna
serratifolia are presented under appropriate heads.
2.1. Genus Premna: An Overview
An overview of the genus Premna with special reference to its taxonomic history,
classification and distribution in the tropical and subtropical regions is presented in this
section. The genus, from its first report in 1771, was under the confused category due to
its indistinguishable taxonomic characters and diverse morphological features. Presently,
Review of Literature Chapter 2

it is one of the largest genera of the family Verbenaceae with two hundred species
worldwide which are mainly distributed in tropical and subtropical Asia, Africa,
Australia and the Pacific Islands (Harley et al., 2004). As this research work is
concerned with Premna serratifolia (type species of the genus Premna), it is relevant to
have a birds eye view of the genus and its taxonomic history.
2.1.1. Taxonomic History of the Genus Premna
The generic name Premna was derived from the Greek word Premnon meaning
tree stump, referring to the short and twisted tree trunks of the type species. Linnaeus
(1771) described the genus Premna with two species, viz., Premna serratifolia and
Premna integrifolia, the types of which were collected by Paul Hermann from Ceylon
and kept in Didynamia Angiosperma. This treatment was followed by Murray (1774),
Gmelin (1791), Willdenow (1800), Sprengel (1825), Roxburgh (1832), Blanco (1837)
and Dietrich (1843). Scopoli (1777) placed the genus in Personatae, which was later
recorded in Centuria Quarta by Gaertner (1788), in Vitices by Jussieu (1789), in
Plasyrgophyta by Necker (1790) and under the tribe Verbeneae of Labiatae by
Reichenbach (1828). In the year 1806, Jussieu referred it to the family, Verbenaceae
where it has been retained by majority of the botanists (Munir, 1984).
Dumortier (1829) was the first author to assign the genus Premna to a tribe,
Viticeae. In 1830, Bartling split Verbenaceae into two sections viz., Viticea and
Verbenea, and the genus Premna was kept in Viticea. Endlicher (1841) divided the
family into three tribes: Lippieae, Lantaneae and Aegiphileae and the genus Premna
was included in the tribe Lantaneae. In 1847, Schauer classified the family Verbenaceae
into three tribes: Verbeneae, Viticeae and Avicennieae, with the genus Premna in the
tribe Viticeae. The genus Premna was retained in this new tribe by majority of botanists
such as Bentham (1870), Bentham & Hooker (1876), Hooker (1885), King and Gamble
(1909) and Fletcher (1938). Schauer (1847) subdivided the tribe Viticeae into three
subtribes: Symphoremeae, Caryopterideae and Viticeae, with Premna in the subtribe,
15

Viticeae. He further split the genus Premna into two sections, viz., Guimira and
Premnos, based chiefly on their calyx being regularly 4 or 5 toothed and also based on
the nature of corolla. Miquel (1858) and Bentham (1876) accepted the above
classification proposed by Schauer (Munir, 1984; Rajendran and Daniel, 2002).
Briquet (1895) while reclassifying Verbenaceae elevated the tribe Viticeae to a
sub-family Viticoidea with four tribes viz., Callicarpeae, Tectoneae, Vitaceae and
Clerodendreae and kept Premna in the tribe Viticeae. Authors namely, Dalla Torre and
Harms (1904), Lam (1919), Gardner (1931), Junell (1934), Moldenke (1959, 1971) and
Melchior (1964) followed the above classification of Briquet. Briquet (1895) in his
classification subdivided the genus Premna into five sections viz., Holopremna,
Odontopremna, Gumira, Premnos and Holochiloma, chiefly based on the size and
number of calyx lobes. The above division of the genus Premna was accepted by Dalla
Torre and Harms (1904). However, most of the later taxonomists did not recognize any
intrageneric divisions in Premna (Rajendran and Daniel, 2002).
2.1.1.1. Indian History of the Genus Premna
A brief history of the genus Premna is given in the taxonomic revision of the
Indian Verbenaceae by Rajendran and Daniel (2002). The first report of the genus
Premna (Linnaeus, 1771) was based on the material (Premna serratifolia) collected by
Konig from peninsular India (Nicolson et al., 1988). In the beginning of 18 th century;
Willdenow (1800) described Premna tomentosa based on Kleins collection from India.
Rottler (1803) described Premna corymbosa based on his collection at Tempakkam
near Madras. He also specified about Premna serratifolia in his work. Jussieu (1806)
described a species, Premna flavescens from Madras, which is now treated as a
synonym of Premna tomentosa. Roth (1821), reported Premna mollissima, which is
now treated as a variety under Premna latifolia Roxb. Roxburgh (1832) described 11
species of Premna from India. Graham (1839) reported four species viz., Premna
cordifolia, Premna integrifolia, Premna scandens and Premna nimmoniana from
16

western India. Walpers (1845) described 11 species of Premna from India in his
monograph. In another study, Schauer (1847) included 25 species from India of which 9
were new and they were based on Wallichs numerical list. From peninsular India,
Wight (1849) reported Premna glaberrima as a new species. Clarke (1885) described
Premna bengalensis, Premna coriacea, Premna khasiana, Premna milleflora and
Premna villosa as new species. Parkinson (1922) reported Premna integrifolia and
Premna divaricata from the Andaman Islands. In another study, Rao (1986) reported
the occurrence of Premna coriaceae, Premna parasitica, Premna pubescens and
Premna pyramidatus in Andaman Islands. Deshpande (1961) reported a new species,
Premna resinosa to the Indian flora. According to Santapau and Henry (1973), there are
25 species of Premna in India. However, according to Moldenke (1980), there are 31
species and 11 varieties of Premna in India. In the taxonomic revision of Indian
Verbenaceae, Rajendran and Daniel (2002), recognized 31 species and 6 varieties of
Premna.Of these three species, viz., Premna balakrishnanii, Premna debiana and
Premna mundanthuraiensis are new species reported from India. Recently, Prabhu
Kumar et al. (2013) reported the discovery of a new species Premna rajendranii from
Western Ghats (Chinnar and Madukkarai) of Kerala. Apart from this, a research team
comprising Robi, Augustin, Sasidharan and Udayan (2013) rediscovered an endemic
and rare species of Premna viz., Premna paucinervis (C. B. Clarke) Gamble from the
Vagamon hills along South Western ghats of Kerala after a lapse of 140 years of its
original type collection by R.H. Beddome (1872) from Anamalayas, Western Ghats
(Tamilnadu).
2.1.1.2. Perplexity in Classification of the Genus Premna
Perplexity in taxonomic classification usually occurs due to the usage of vague
characters as distinguishing features. In taxonomic history of Premna, such a confusing
situation occurred by the separation and consequent merging of genus Pygmaeopremna
with genus Premna. In 1910, Merrill described a new genus Pygmaeopremna, based on
two collections from Luzon, Philippines. The type species was named Premna humilis
17

Merr. and the genus was related to Premna and Vitex, and have shown
more
resemblance to the genus Premna. The genus Pygmaeopremna was distinguished from
Premna only by its very small size. Lam (1919) recognized Pygmaeopremna as a valid
genus, but at the end of his generic description noted: without regard to characteristics
of less consequence, the genus differs from Premna only by its extraordinary small size;
therefore perhaps we had better combine it with the genus. Further in 1921, Lam &
Bakhuizen suggested that Pygmaeopremna could not be distinguished from Premna
L. Based on taxonomic investigations, Munir (1984) stated that the flowers of all
available Pygmaeopremna collections were similar to those of Premna. The only
differential character that really holds is the dwarf habit of Pygmeopremna, and this
character is not good enough to recognize the genus. Hence, following Merrill (1910,
1923, 1951), Pygmaeopremna is now considered as a synonym of Premna (Munir,
1984).
2.1.2. Distribution of the Genus Premna
The genus Premna L. now contains about 200 species worldwide which are
mainly distributed in the tropical and subtropical regions of Asia, Africa, Australia and
the main distribution extends from India to Japan, Southward to Indochina, Malaysia,
and tropical Australia and eastward to Polynesia (Munir, 1984; Kirtikar and Basu,1992;
Harley et al., 2004; The Forest Herbarium, 2001; Kadareit, 2004). A study of the genus
in Thailand was first undertaken by Fletcher (1938), enumerating 30 species. Later,
Moldenke (1980) and Govaerts et al., (2008) reported the checklists of Premna with 39,
19 and 33 taxa, respectively. In another study, Rajendran and Daniel (2002), reported
31 species and 6 varieties of Premna from India.
2.2. Premna serratifolia: An Overview
An overview of Premna serratifolia with special reference to its taxonomic history
(nomenclature, species complexity & synonyms, polymorphic status and geographic
distribution) is described in this section.
18

2.2.1. Taxonomic History of Premna serratifolia
The type species of the genus (generic type) for the genus Premna is Premna
serratifolia L. The nomenclature and identity of this widespread and very polymorphic
species has been subjected for much heated discussions and negotiations among
taxonomists (Merrill, 1917; Fletcher, 1936; Meeuse, 1942; Fosberg, 1953). Many
authors have given different names viz., Premna integrifolia L., Premna serratifolia L.,
Premna corymbosa (Burm.f.) Merr., Cornutia corymbosa Burm.f. for this species
complex. Of the above mentioned species, the type of Premna integrifolia was collected
by Paul Hermann in Ceylon. This species was first described by J. Burman (1737) under
the name Sambucus zeylanica odorata aromatica and Linnaeus (1747) renamed it
using a single name Cornutioides. Later, N. L. Burman (1768) for the first time
renamed this Ceylon material by giving a binomial name, Cornutia corymbosa.
However, Linnaeus (1771) considering the complexity of the Ceylon material
being a mixture of more than one taxon, divided this into two species viz., Premna
integrifolia and Premna serratifolia based on the morphology of their leaf margin being
entire and serrate respectively. Linnaeus cited under Premna integrifolia N.L. Burmans
validly published binomial Cornutia corymbosa, thus making the binomial Premna
integrifolia an illegitimate name (Art67, Int.code Bot.Nom. 1978). Linnaeus cited his
invalid single name Cornutioides Fl.Zeyl.416 under the plant material with serrated
leaf margin, Premna serratifolia. According to Linnaeus, the serrate leaved plant in
Hermanns Ceylon material was different from Burmans Cornutia corymbosa. Since
the serrate-leaved segregate of the Ceylon material, on which Premna serratifolia was
based, did not involve the entire leaved type material of Cornutia corymbosa Burm.f.,
and would seem that Premna serratifolia is a legitimate name. According to Lourteig
(1966), the type of Cornutia corymbosa Burm.f., on which Premna integrifolia was
based, is preserved at the Institute de France, Paris (Munir, 1984). Based on the above
discussions, it can be concluded that taxonomically Premna integrifolia and Premna
serratifolia belongs to the same species, but nomenclaturally they are based on two
19

different types, segregated from Paul Hermanns Ceylon material. Schauer (1847)
pointed out these two as synonymous and hence united these two species under the
name Premna serratifolia L.
Rottler and Willdenow in 1803 described one of Rottlers collections from
Madras, India, as a new Premna species, Premna corymbosa. In the protologue, they
mentioned this as Premna corymbosa Nob., making no reference to Cornutia
corymbosa Burm.f. Rottler contrasted Premna corymbosa with Premna integrifolia L.
and in a footnote to his paper, Willdenow contrasted three other species of Premna with
Premna corymbosa, without mentioning Burmans Cornutia corymbosa. Here also,
taxonomically Premna corymbosa Rottl. & Willd. and Cornutia corymbosa Burm.f. are
identical, but nomenclaturally Premna corymbosa Rottl. & Willd. was based on a
specimen from India while Cornutia corymbosa Burm.f. on a specimen from Ceylon
respectively. Therefore, the combination of Premna corymbosa (Merrill, 1917) based
on Cornutia corymbosa Burm.f would be a later homonym and illegitimate. In 1917,
Merrill clearly mentioned that Premna corymbosa (Burm.f.) Rottl. & Willd. is the
correct name for the plant that Linnaeus named Premna integrifolia. He further pointed
out that all three (i.e. Premna integrifolia, Premna serratifolia and Premna
corymbosa) are typified by the same material. This view was later accepted by
Fletcher (1936) and Meeuse (1942). Howevever, Fosberg (1953) disagree with the
above argument and pointed out that Premna corymbosa Rottl. & Willd. (1803) was
described as a new species from India and it was not based on Cornutia corymbosa
Burm.f., the type of which came from Ceylon. Nevertheless, Fosberg (1953) did not
appreciate the fact that Paul Hermanns Ceylon material, on which Corntia corymbosa
Burm.f was based, had been treated by Linnaeus (1771) as a mixed collection; he
considered that both names were based on the type of Corntia corymbosa Burm.f., and
were therefore, illegitimate. The fact that Premna serratifolia was differently typified
and therefore legitimate have been missed by Fosberg (1953) and Moldenke (1971,
1980). They therefore, accepted Premna obtusifolia R. Br. (1810) which they
considered to be the next available name for this species. As it is evident from the above
20

discussion, Premna serratifolia L. (1771) is the oldest legitimate name in this species
complex for Premna, which comprises Premna serratifolia L., Premna integrifolia L.,
Premna corymbosa Rottl. & Willd., Cornutia corymbosa Burm.f. and Premna
obtusifolia R.Br.
Currently, Premna serratifolia L. is accepted as the oldest validly published
name and all other names viz., Premna integrifolia L., Premna obtusifolia R.Br. and
Premna corymbosa (Burm.f.) Rottl. & Willd. are treated as its synonyms (Munir,
1984).
2.2.2 Distribution of Premna serratifolia
This species is seen distributed in Papua New Guinea, New Britain, New Ireland,
East Africa, India, Southern China, Java, Moluccas, Malaysia, Philippines, Samoa, Fiji,
Indochina, Australia and Japan. According to Lam (1919) the species is seen distributed
from Madagascar, Mauritius, India to Malacca and Thailand, East Bengal, Ceylon,
Andaman, Nicobar, Hong Kong, Malaya, Philippines and Polynesia. Apart from these
localities Moldenke in 1971 recorded it from Hainan, Taiwan, Ryukyu Archipelago and
Melanesia.
In India, it is distributed in Assam, Khasi hills, Goa, Gujarat, Karnataka, Kerala,
Lakshadweep, Maharashtra, Orissa, Tamilnadu, West Bengal and coastal forests of
Andaman and Nicobar Islands (Kurz, 1974; Hooker, 1985; Rajendran and Daniel, 2002;
Khare, 2007). According to Gamble (1967) the species is seen distributed in Deccan
forests, Kodur in Cuddapah, Nellore, Chingleput and Madras to the Javadis near the
coast. It is reported from Sylhet and Coromandel of Bombay, South Kanara, South
Carnatic and Travancore-Cochin, where it grows under saline and mesophytic
conditions (Chopra et al., 1956; Somasundaram, 1963; Drury, 1858; Pharmacognosy of
Ayurvedic drugs, 1978; Varier, 1995). The plant is common in many parts of India
especially towards sea-coasts (Parkinson, 1922; Chopra and Chopra, 1955; Nairne
21

1976; Bapalal Vaidya, 1982; Manilal and Sivarajan 1982; Kartick, 1984; Sivarajan and
Indira, 1996; Nadkarni, 2007).
2.2.3. Polymorphic status of Premna serratifolia
The genus Premna was considered as an extremely difficult genus in which
flower features are almost as vague and little distinct as those of the extremely variable
leaves (Beer & Lam, 1936). As Bentham (1870) rightly pointed out there are a
number of forms including Premna integrifolia and Premna serratifolia of Linnaeus
which seem to pass into each other by numerous intermediates, and it would require a
much more detailed study of good specimens from different localities. Earlier
taxonomists like Schauer (1847), Schumann and Hollrung (1889) and H.J. Lam (1919)
stressed the importance of merging of related species into a large and very polymorphic
one, being called by the name of eldest, probably, Premna integrifolia. H.J. Lam
(1919) regarded Premna integrifolia as a very polymorphic species and stated that
for examining a large number of specimens, we found, that several other species were
inseparably united with one another by all possible intermediate forms and with Premna
integrifolia.. According to Kok (2013), Premna serratifolia shows significant
morphological variation across a wide geographical distribution, especially in leaf
shape, leaf margin, inflorescence size and calyx form. This was recognized and well
illustrated by Lam (1919) and Munir (1984). Nevertheless, Lam (1919) accepted that
two subspecies could be recognized viz., Premna integrifolia sub sp. truncatolabium
and Premna integrifolia sub sp. dentatolabium mainly based on the leaf shape. After
studying the specimens available, Lam & Bakhuizen van den Brink (1921) decided that
these sub species could no longer be recognized, and instead suggested five different
types: type I- integrifolia, type II- abbreviate, type III- cyclophylla, type IV- sambucina
and type V- foetida. These subdivisions were suggested based on a series of continuous
characters and ecological data (habitat; from seashore to inland; from shrub to tree;
lamina becoming bigger and more acuminate and calyx becoming more distinct and
toothed) which, according to the authors, overlapped and formed a cline. In 1919, H.J.
22

Lam expressed the difficulty of subdividing the species exclusively based upon the form
of calyx, which, as in some other species, is often inconstant/ variable. He also
commended on the wrong and unscientific way to discover the truths using old methods
of mere morphological examination and recommended the application of new branches
of science like genetics to solve the problems in systematic botany. In spite of such
serious attempts to study the wide range of variations in Premna serratifolia, no solid
evidences have been generated either to recognize or differentiate the different morphovariants of this taxon. However, when an extensive amount of material is examined the
variation is found to be continuous and might have resulted by the process of gradual
evolutionary changes.
2.3. Premna serratifolia: A potential Medicinal plant
Premna serratifolia L. is a well-known medicinal plant belonging to the family
Verbenaceae. Regardless of its morphological variations, this polymorphic species has a
unique status in traditional systems of medicines like Ayurveda and folklore medicines.
A brief review of literature pertaining to its traditional medicinal uses and traditional
medicine preparations are summarized below.
2.3.1. Traditional Medicinal uses of Premna serratifolia
The indigenous medicinal properties of Premna serratifolia have been reported
by many Indian researchers (Chopra et al., 1956; Chopra, 1969; Nadkarni, 1976;
Rathore et al., 1977; Kartick, 1984; Rastogi and Mehrotra, 1991; Warrier et al., 1994;
Lalithamma, 1996; Yoganarasimhan, 2000; George and Samuel, 2003; Thomas and
George, 2005; George et al., 2006; Prajapati et al., 2006). As reported by Kartick
(1984) and Sivarajan and Indira (1996), its root, stem/ stem bark and leaves are used for
different traditional preparations in medicine. Premna serratifolia, also known by the
name Agnimantha, is acrid, bitter, astringent, cardio tonic, carminative, alterative,
laxative, stomachic and tonic. It improves digestive power and is useful in constipation,
fever, heart diseases and neurological diseases. It overcomes kapha and vata disorders,
23

anemia, piles, oedema, poison, anasarca and abdominal diseases. Traditionally, the drug
is highly valued for its anti-inflammatory property (Kolammal, 1979; Kurup et al.,
1979). It is pungent and hot in action and used to treat weak digestion and excessive
fatty condition of the body (Pharmacognosy of Ayurvedic drugs, 1978). An
ethnobotanical study conducted among traditional Ayurvedic vaidyans of Kerala by
George (2006), revealed many hitherto unreported indigenous uses of Premna
serratifolia. According to this study, the leaves have more medicinal applications
(55.55%) followed by roots (36.11%) and bark (8.34%). As reported in this study,
Premna serratifolia is specific to anemia, weak digestion, piles, constipation,
amavatam, accumulation of fluid in the tissues or in body cavities and also effective to
reduce serum cholesterol level and eliminating poison/ heavy metals from the body
parts. Premna serratifolia is traditionally used in the treatment for beriberi, vaginal
irritation, to relieve headache and as eye lotion (Dassanayake and Fosbergeds, 1980;
Padua, 1999).
Traditional healers consider leaf as one of the useful parts for internal as well as
external uses. The leaves are galactagogue, and are useful in agalactia, catarrah,
rheumatalgia and tumours (Varier, 1995). Leaves are used as an external application to
piles and tumours (Ayurveda). Leaf decoction of the species is used for bathing infants.
Leaves are also used for the treatment of arshas (hemorrhoids) in the form of Avagaha
sweda (sitz bath) to reduce inflammation and pain. Sugared decoction of Premna
serratifolia leaves with the juice of calamansi (a citrus fruit) helps to loosen up phlegm
and effective for coughs. Decoction of leaves is used as curative medicine for fever,
blisters of the lips and stomach aches (Sturtevant, 1972). Leaves when applied over the
urinary bladder facilitate urination (Kumar and Jain, 2002). The leaves are eaten by the
inhabitants of the Coromandel Coast (Whitelaw, 1984). In Peninsular Malaysia and
Indonesia, boiled young leaves are consumed as vegetable. The leaves are also used for
the preparation of natural jin and sherri (Hussain et al., 1992; Steven, 2005).The
leaf is diuretic and indicated in dropsy (Agarwal, 1997). The leaf decoction is reported
to be good for the treatment of colic, rheumatism and neuralgia (The Wealth of India,
24

1948). The plant is mentioned as appel in Hortus Malabaricus by Rheede (1678-1693),
and the decoction of' its leaves is suggested for pains and wind in the stomach.
According to Burkill (1966), specifically the leaves of Premna obtusifolia are used in
folklore medicines. George (2006) also reported that, of the four types of Premna
serratifolia, the leaves of Neelan munja are effective for marma treatment, neuralgia
and as an analgesic to treat vata disorders. The tender plant is used for neuralgia and
rheumatism. Leaves rubbed along with pepper administered in colds and fevers; in the
form of a decoction given for flatulence; in form of soup used as stomachic and
carminative. Infusion of the leaves (1 in 10) is used in eruptive fevers, colic and
flatulence, in doses of 1 to 2 ounces (Nadkarni, 2007). The whole plant is given in
decoction for pains in the head and body, as well as, in rheumatism. This plant is an
essential ingredient of Medhahara kwata which is often prescribed by traditional
vaidyans for obesity. According to George (2006), the leaf extract of Premna
serratifolia can be effectively utilized to evolve an eco-friendly pest-management
strategy to control chicken fleas, coconut pest (Oryctes rhinoceros) and a variety of
vegetable pests.
The root has an agreeable aromatic odour (Dey, 1980). The root forms an
ingredient of dasamula, a preparation often prescribed by the native physicians in
obstinate fevers (Kirtikar and Basu, 1992). The root is pungent, useful in chyluria, and
swellings. It is thermogenic, anodyne, alexeteric, expectorant, depurative, febrifuge and
antibacterial. The roots are useful in cardiac disorders, hepatopathy, cough, asthma,
bronchitis, leprosy, migraine, jaundice, malaria, skin diseases, dyspepsia, inguinal
hernia, kidney stones, rhinitis, rheumatoid arthritis, abscess haemorrhoids and is also
indicated in shotha (inflammation), vatavyadhi (neurological disorders), prameha
(diabetes mellitus), medoroga (obesity) and agnimandya (loss of appetite). In Unani,
the root is suggested good for liver complaints. The species is known to have various
folklore usages (Nadkarni, 1985; The Wealth of India, 2005; Fairchild, 1943; USDA,
2007). This root is prescribed in decoction, as a gentle cordial and stomachic in fevers.
For fevers, a quantity of half a tea-cupful, twice daily is prescribed. Decoction of root is
25

good for liver complaint. Decoction of the root (1 in 10) or about 4 ounces in a pint of
water and boiled for 15 minutes, is given in doses of 2 to 4 ounces twice daily as a
stomachic and a tonic, and also in gonorrhea and during convalescence from fevers.
Leaves are also used for the same purpose. Root forms an ingredient of Dasamula and
thus used in a variety of afflictions. Root rubbed into a paste with water is
recommended to be taken with clarified butter in urticaria and roseola for a week
(Nadkarni, 2007). The root boiled in salt water is used in gout, which is externally used
(Drury, 1858). Traditionally, it is used in bone fractures and also to reduce pains in
bones and rheumatic aches. In various parts of Indonesia, an infusion of the leaves and
roots are used against fevers and shortness of breath and to promote breast-milk
production in women. In Indo-China, the leaves and roots are used in traditional
medicine as a diuretic, stomachic and febrifuge (Hussain et al., 1992).
Decoction of stem bark decreases force of contraction of heart and produces
dilation of the pupils (Chopra et al., 1956). The alkaloids in the species have a
sympathomimetic action (Chopra and Chopra, 1955). On Guam, in the Pacific Ocean, a
tea made from the boiled bark is used to treat neuralgia. It is very effective for the
treatment of kidney disease, liver problems, and constipation (Hussain et al., 1992).
2.3.2. Premna serratifolia in Ayurvedic formulations
Premna serratifolia is one of the drugs that constitute the drug groups (gana)
such as Dasamula, Brahatpanchamula, Viratarvadi, Varunadi gana (Sushruta);
Sothahara, Sitaprasamana and Anuvasanopaga (Charaka) (Pandey, 2002). In Ayurveda
various drug formulations like arishtam, rasayanam, kwatham, ghritham, and thailam
are used as a part of its treatment strategy. In Arishtams (naturally fermented herbal
decoctions)
such as Amritharishtam,
Dandyarishtam,
Dasamoolarishtam
and
Balarishtam, Premna serratifolia is used as a major constituent. The major rasayanams

(nutritional juices of medicinal plants) having Premna serratifolia as an essential
constituent are Agasthyarasayana, Chyavanaprasam, Dasamoolarasayanam and
26

Brahmarasayanam. In Ayurvedic medicines, kwatham is an unfermentated form of
herbal decoction prepared by adding water in specified proportion to the coarsely
powdered herbal material followed by further boiling and filtering processes. The main
kwathams prepared
from the
powdered roots of Premna
serratifolia
are
Indukanthakwatham, Dhanwantharakwatham, Varanadikwatham, Sapthasarakwatham,

Medhahara kwatham, Maharasnadi kwatham and Luhuna Kolla kwatham.Ghritham are
ghee preparations of plant extracts used for internal consumption. Inducantha ghritham,
Sukumara ghritham, Dhanwanthara ghritham, Dasamoola shatpala ghritham and
Medhahara ghritham are some of the major ghrithams containing Premna serratifolia
as major ingredient. Thailams are mainly the oil preparations having herbal extracts
which is usually used for external applications. Dhanwanthara thailam, Prabhanjana
thailam, Vimordhana thailam, Narayana tailam and Sahacharadi thailam are prepared
with poly herbal extracts containing Premna serratifolia as a key constituent (API,
2001). Premna serratifolia is also used in preparations like Dasamoola hareetaki,
Agnimantha-mulkalka and Agnimantha-kasaya (Dey, 1980).
2.3.3. Description of Premna serratifolia under Sanskrit names in Traditional
Manuscripts
Premna serratifolia is described in the traditional Ayurvedic manuscripts under
many Sanskrit and vernacular names depicting its morphological and diverse medicinal/
therapeutic uses. The most commonly used Sanskrit vernacular name attributed to this
medicinal plant is Agnimantha. The name Agnimantha is derived from its use during
Vedic period, where its stem/ sticks were used to produce fire (The Wealth of India,
1972). Other Sanskrit names of Agnimantha viz., Vahnimantha and Havirmantha also
indicates, that the tree was used to produce fire in the sacrificial ceremonies by rubbing
the sticks together (Sivarajan and Indira, 1996).
The different uses of Agnimantha were mentioned in Rig-Veda and AtharvaVeda. In Astanga Sangraha, the plant is described by the name Arani which is the
synonym of Agnimantha. The different medicinal properties of Agnimantha were
27

described in Laghutrayis. In Nighantukaras, besides medicinal properties, its
morphological characters were explained in detail. The major manuscripts having the
description of this species are: Bhavaprakasa, Saligrama Nighantu, Dhanwanthari
Nighantu, Madanapala Nighantu, Nighantu Ratnakara, Abhidhanamanjari, and Raja
Nighantu (Pharmacognosy of Ayurvedic drugs, 1978). In these traditional Sanskrit texts,
many synonyms such as Agnibijaka, Agnimantharkari, Ananta, Arani, Aranika,
Araniketu,
Ganakasika,
Ganikaarikaa,
Ganikasika,
Gankarika,
Girikarnika,
Harimantha, Havirmantha, Jaya, Jayanthi, Jayee, Jyotishka, Kanika, Karnika, Kethu,

Kshudragnimantha, Manthanam, Mathana, Nadeyi, Nadija, Nathy, Pavaka, Pavakarini,
Pittamata, Rakthangam, Shriparna, Shriparni, Tanutvaka, Tarkari, Tejomantha,
Vaataghni, Vahnimantha, Vaijayantika, Vanhimula, Vijaya, Vijayantika and Vyganthika
were given to Agnimantha to describe either its medicinal attributes/ properties or its
morphological features.
Premna serratifolia is also known by different names in different states of India.
Its common names in different regional languages are as follows.
Hindi
: Arni, Agetha, Arani, Ganiari
Tamil (Tamilnadu)
: Munnai, Munnay, Munney, Munni, Munni-vayz, Pasumunnai,

Peyminnay.
Telugu (A.P.)
: Gabbunelli, Ghebunelli, Kanika, Karnika, Nagura, Nelli,

Nelli chettu, Padmaka, Pinnanelli, Tukkadu.
Malayalam (Kerala) : Munja, Munna, Appa, Appel, Kozhychedy.

Kanada (Karnataka) : Agnimandha, Naravalu, Takkila.
2.3.4. Controversial status of Premna serratifolia in Ayurveda
In the Ayurvedic system, there is considerable disagreement regarding the
identity of genuine medicinal plants since many of the drug plants are not described
with scientific precision in classic Ayurvedic texts. Hence, many unauthentic plants or
28

plant parts are being used for the preparation of standard classic Ayurvedic medicine in
different places and sometimes even in the same locality. Premna serratifolia, is one
such medicinal plant coming under controversial drug plants of India. The review of
related literature revealed that there is much disagreement among the commentators of
modem Nighantus regarding the identity of genuine drug, Agnimantha. When a few
scholars did not refer to different source plants for Agnimantha, majority of the
Ayurvedic scholars (Purandare,1896; Sthana, 1916; Acharya, 1950; Nadkarni et al.,
1954; Vaidya,1965; Krishnamurthy et al.,1972; Meulenbeld,1974; Chaturvedi et
al.,1983; Jain, 1986; Handa and Kaul, 1996; Sharma, 1996; Meulenbeld and Wujastyk,
2001; Puri, 2002; Bishnupriya et al., 2003; Nair, 2004) raised doubts regarding the
identity of genuine source drug of Agnimantha.
A very serious limitation of the Ayurvedic system is the difficulty in ascertaining
the identity of the genuine medicinal plants prescribed by the founders of the system. In
the original texts, the descriptions of the medicinal plants were given in poetic language
and hence very often, lack scientific precision. Hence, the interpretation of the
description in Sanskrit is largely influenced by the views of the interpreter. A short
review of the views of different authors is compiled below.
Two types of Agnimantha viz., Laghu (small) and Brihat (big) having somewhat
similar properties were mentioned in Nighantus. Charaka and Sushrutha have
mentioned them separately as Agnimantha and Tarkari respectively. In Sushrutha
Samhita, Agnimantha is mentioned as one among Brihat panchamula and it is equated
to Premna integrifolia by many commentators. Sushruta while explaining Varunadi
gana, Tarkari and Agnimantha were mentioned separately, indicating these two as
separate drugs. In Charaka Samhitha also has mentioned about two varieties viz.,
Agnimantha and Tarkari and are described as separate trees. However, Charaka
described these two, Agnimantha and Tarkari, together in a similar context. Later
authors have equated Clerodendrum phlomidis to Laghu and Premna corymbosa
29

(Premna serratifolia, Premna integrifolia) to Brihat as the respective sources of the two
varieties of Agnimantha (Bapalal Vaidya, 1982; Chunekar, 1982).
The texts and lexicons of Ayurveda mentioned about two varieties
of Agnimantha, namely Brihat Agnimantha (big variety) and Laghu Agnimantha or
Ksudragnimantha
(small
variety)
which
have
been
correlated
to Premna
integrifolia and Clerodendrum phlomidis respectively by many scholars. While writing

vimarsha for the sloka,Tarkari dwayam' in Ashtanga samgraha, the commentator
explains that Agnimantha is of two varieties Tarkari and Agnimantha; Tarkari is also
called as Laghu Arani and it is botanically equated to Premna integrifolia, while
Agnimantha is called as Vruddha Arani and is botanically equated to Clerodendrum
phlomidis. Among the Nighantus, Amarakosha considered Agnimantha and Tarkari as
different plants. Sodhala (1994) mentioned Tarkari and Agnimantha (Arani) as two
different varieties (Clerodendrum phlomidis and Premna integrifolia as the botanical
sources of Tarkari and Agnimantha respectively). Some of the modern time scholars of
Dravyaguna consider Tarkari as Laghu Agnimantha. Bhavamishra mentioned only one
variety i.e., Agnimantha for which Tarkari is the synonym. In Nighantu Ratnakara,
Agnimantha is described as two types viz., Laghu (smaller) and Brihat (bigger). This
text explains that Laghu variety has better sothahara property than the Brihat variety.
Kaiyyadeva Nighantu also quoted Agnimantha as a better sothahara and vatahara drug.
In Dhanwanthari Nighantu two varieties of Agnimantha are mentioned, Agnimantha
and Kshudragnimantha or Kshuragnimantha. Commentator considered Clerodendrum
phlomidis as Agnimantha and Premna integrifolia as Kshudragnimantha. In Priya
Nighantu, Arani and Agnimantha are used synonymously. The commentator considered
Premna integrifolia as Agnimantha. In Raja Nighantu and Saligrama Nighantu, two
varieties are mentioned: Agnimantha and Kshudragnimantha. Both these are mentioned
under Prabhadradivarga in Raja Nighantu. The synonyms of Agnimantha are also
given for Jayanti, included under Satahvadivarga. For these the commentators cite the
Latin name Clerodendrum phlomidis, though Tarkari is regarded as a synonym of
Agnimantha. According to Pandey (1998), Agnimantha is of two types viz., Brihat
30

Agnimantha and Kshudragnimantha and are equated to Clerodendrum phlomidis and
Premna integrifolia respectively.
Unscientific nomenclature of medicinal plants followed in classical traditional
Sanskrit texts is a serious defect of Ayurvedic system. In Ayurveda, unlike modern
botany, there is no precise and uniform system of nomenclature. Dozens of names may
be found given to one and the same plant, each name indicative of one minor attribute
or diagnostic property of the plant. The loose, intuitive and unscientific way by which
ancient authors have named plants is the source of much confusion today, because the
qualitative names are very often applicable to more than one plant. Similarly, there are
also cases of different species of plants (taxonomically related or unrelated species)
having common medicinal properties, owing to the presence of same organic
compounds. In the case of Agnimantha, the two species suggested are Premna
serratifolia and Clerodendrum phlomidis, which are botanically closely related species
belonging to the same family Verbenaceae (Singh et al., 1972). In South India,
particularly in the state of Kerala, Premna serratifolia has been used as the genuine
drug. However, in North India Clerodendrum phlomidis is used instead of Premna
serratifolia. The properties attributed to these two species are considered to be the
same. The property of Laghu Agnimantha is same as Brihat Agnimantha but Laghu is
better for lepa and poultice in swelling.
Most of the regional names of Agnimantha such as Gineri, Agethu, Tekara or
Tankali are actually distorted forms of the original Sanskrit names- Ganikarika,
Agnivadhu and Tarkari. There is not much difference in the tree sizes of the two kinds
of Agnimantha and thus any attempt to differentiate them as Vrhat (large) and Ksudra
(small) kinds appears untenable. Their separation on the basis of plant size may,
however, be limited to different species of Premna only. It is, however, reasonable and
useful for the sake of field identification to name the Premna species as
Ksuragnimantha (thorny Agnimantha) and Clerodendrum species as Aksuragnimantha
(without thorns). Arani and Agnimantha may be treated as common names for both the
31

species but Tarkari may be accepted as a permanent name for Clerodendrum species.
The justification for accepting the other names as synonyms for Agnimantha and its
varieties has been discussed based on their properties and uses. It may be noted that the
differences in uses and properties of the two varieties have not been pointed out either
in the Nighantus or in the preparations (Yogas) of the texts. Instances are not inadequate
where Agnimantha and Tarkari have been treated as equivalents in the identical Yogas
of different texts.
Several authors have correlated Agnimantha to other species of the genus
Premna. Kolammal (1979) and Nair (2004) correlate Agnimantha to Premna
serratifolia L. and Premna latifolia Roxb. as a substitute. Warrier et al. (1994) and
Sivarajan and Indira (1996) correlate Agnimantha to Premna corymbosa Rottl. Sharma
(2006) mentions two varieties: Agnimantha (bigger variety) correlated to Premna
mucronata Roxb. and Tarkari (smaller variety) equated to Clerodendrum phlomidis.
Premna serratifolia L. and Premna spinosa Roxb. are the other species correlated to
Agnimantha. Premna latifolia Roxb. var. mucronata C.B.Clarke (a botanical synonym
of Premna
mucronata Roxb.)
has
also
been
considered
by
some
authors
as Agnimantha.
In most of the traditional texts of Kerala except Bhavaprakasham and
Madanapala Nighantu, two types of Munna are mentioned and their synonyms are
often used interchangeably. The two types of Munna mentioned in Madanapala
Nighantu are Munna and Kattumunna. P.V. Sharma (1998) in Dravyagunavinjan has
considered Agnimantha to be Valiya munna (Premna mucronata) and Tarkari to be
Ceriya munna (Clerodendrum phlomidis). However, throughout Kerala Premna species
is used for Agnimantha and Tarkari (Varier, 1995). In Ayurveda Vishwa Vijnan Kosham
three types of Munja viz., Puzhamunja, Munja and Cherumunja were described.
There are various views regarding the genuine source plant and substitute of
Agnimantha. According to Data base on medicinal plants used in Ayurveda,
32

Clerodendrum phlomidis is considered as the genuine drug and Premna serratifolia as
the substitute since these two plants have similar medicinal properties (i.e., in Guna
karma they are treated identical). Chunekar (1982) in his commentary on
Bhavaprakasa Nighantu also opined these two as substitutes for each other, since they
have similar medicinal properties. A few authors like Nadkarni (1976), Kamat and
Mahajan (1972) pointed out Clerodendrum inerme (L.) Gaertn. as the source plant of
Agnimantha. However, majority of the authors like Kirtikar and Basu (1918), Vaidya
(1936), Kurup et al. (1979), Dey (1980) and Mooss (1980) equate the drug with Premna
serratifolia L.
In the first edition (Part-I) of Ayurvedic Formulatory of India Clerodendrum
phlomidis Linn.f. is mentioned as the authentic botanical source and Premna
integrifolia Linn. and Premna mucronata Roxb. as a substitute (AFI, 1978). In its
second edition (Part-I), Premna integrifolia has been mentioned as the authentic plant
source and Clerodendrum phlomidis and Premna mucronata as the substitutes (AFI,
2000)a. In the first edition (Part-II) of Ayurvedic Formulatory of India also
Clerodendrum phlomidis. Linn.f. has been listed as the authentic Agnimantha and
instead of Premna integrifolia Linn., Premna obtusifolia R. Br. and
Premna
mucronata Roxb. are listed as the substitutes (AFI, 2000)b. As Aparna et al. (2012),
rightly pointed out, the basis for this variation in listing of botanical sources
for Agnimantha are not provided and hence not clear.
As it is evident from the above discussions, it is very difficult to resolve the
controversy regarding the identity of Agnimantha as the available evidences are
inadequate to equate this drug plant to either Clerodendrum phlomidis L./
Clerodendrum. inerme (L.) Gaertn. or Premna serratifolia L. Based on the available
evidences listed in classical texts and Nighantus, it is not easy to judge the authenticity
and exact identity of the source drug Agnimantha. Based on comparative morphological
characters described in classical texts, it is quite likely that both plants (Premna
serratifolia and Clerodendrum phlomidis/ Clerodendrum inerme might have identical
33

therapeutic properties as evidenced from their time tested ethno medical uses. Marked
discrepancy in pharmacognostical parameters of these plants were reported by different
authors. However, it is expected that in depth studies based on modern tools and
techniques in pharmacognosy and phytochemistry will resolve the existing ambiguity
and may provide additional evidences for establishing the identity and purity of the real
source plant.
2.4. Phamacognostic, Phytochemical and Pharmacological studies on
Premna serratifolia
A brief report of the studies done with respect to Phamacognostic, Phytochemical
and Pharmacological studies of Premna serratifolia L. is presented below under
appropriate heads.
2.4.1. Pharmacognostic studies on Premna serratifolia
Pharmacognosy is one of the indispensable tools in distinguishing medicinally
important plants from its substitute or adulterant counter parts. In botanical
pharmacognosy prime importance is given to the botanical aspects of the plant and its
growing habitat, harvesting and processing conditions rather than for its chemistry for
ensuring the plants medicinal efficiency. The interest in traditional herbal medicines is
reemerging worldwide, precisely because many modern drugs, whether synthetic or
derived from nature, are failing to serve the health care needs of majority of the world
population. The classical botanical pharmacognosy mainly deals with and concentrates
in maintaining the quality of the plant, the environment in which it grew, and its myriad
compounds and actions that are of importance and most appropriate in the development
of traditional herbal medicines that people worldwide rely upon in self healing and
traditional healing systems. The pharmacognostic studies carried out with special
reference to Premna serratifolia are summarized below.
Studies on pharmacognostic, ethnobotanical and phytochemical aspects of
different ecotypes of Premna serratifolia in Kerala were initiated under the leadership
34

of George in the year 2003, in Department of Botany, C.M.S. College, Kottayam. He
conducted an extensive survey among the traditional Ayurvedic practitioners of Kerala.
He collected many morpho-variants of Premna serratifolia from different regions and
the germplasm of these morpho-variants were maintained in C.M.S. College herbal
garden and also in his experimental home garden at Karunyna lane, Kalathilpady,
Vadavathoor P. O., Kottayam - 686010. A project (funded by UGC) entitled
Ethnobotanical, Phytochemical and Pharmacognostic studies on Premna serratifolia
L. was successfully completed and submitted to University Grants Commission
(George, 2006). In another study entitled Pharmacognostic studies on Agnimantha,
George et al. (2006) identified four different ecotypes of Agnimantha viz., Kozhimunna
(Ecotype-1),
Chemparathimunna
(Ecotype-2),
Cherumunna
(Ecotype-3)
and
Neelanmunna (Ecotype-4) from different parts of Kerala. The study also revealed the
existence of different ecads for Kozhimunna and Chemparathimunna. As revealed in
this study, the existence of different ecotypes and ecads of Premna serratifolia in the
different geographical areas of Kerala shows the high degree of plasticity possessed by
Agnimantha.
The macro and microscopical characters of the roots of Premna integrifolia and
Clerodendrum phlomidis were studied by Gokani et al. (2008). Morphologically the
roots of both species resemble each other except for their colour and size.
Microscopically they can be differentiated by noting the presence of rhytidoma in roots
of Premna integrifolia. Starch grains were found distributed only in the xylem
parenchyma and xylem rays in Clerodendrum phlomidis, whereas in Premna
integrifolia starch grains were detected in all tissues except cork. In another study,
Rajendran and Susheela (2010) conducted a preliminary investigation to standardize
certain pharmacognostical parameters viz., physico-chemical, phytochemical and
fluorescence analysis of stem bark and stem wood of Premna serratifolia collected from
Tamilnadu State. The anatomical features of the root and stem of Premna serratifolia L.
were reported by earlier researchers (Pharmacognosy of Ayurvedic drugs, 1978;
Lalithamma, 1996; George, 2006).
35

Thirumalai et al. (2013) studied the morphological and microscopical characters
of Premna herbacea, a related species of Premna serratifolia. In this study, the roots
and root powder of Premna herbacea were subjected to morphological (colour, odour,
taste, shape and texture) and microscopic structural evaluation. Coarse powder samples
of the plant roots were used to perform physiochemical studies such as total ash, acid
insoluble ash, water soluble ash, extractable matter, loss on drying, foaming index and
swelling index. Root powder samples were treated with different reagents and observed
for fluorescence under visible light and under UV light of short and long wavelength.
They exhibited fluorescence. The physiochemical parameters of the plant were within
the limits. Phytochemical analysis of the root extracts Premna herbacea in different
solvents (ethanol, chloroform, petroleum ether and water) revealed the presence of
triterpenoids and alkaloids with trace amounts of carbohydrates and flavonoids. TLC
and HPTLC analysis of the various extracts also yielded satisfactory results.
2.4.2. Phytochemical studies on Premna serratifolia
Plants serve as the primary sources of medicine from ancient period and the
search for novel plant derived lead molecules of therapeutic importance has gained
momentum in recent years. A brief description of the phytochemical compounds
reported from different parts of Premna serratifolia along with different approaches
adopted for identification, screening, isolation, characterization and in vitro production
of active compounds are summarized in this section.
Major Phytochemical compounds in Premna serratifolia
Many phytochemical compounds were reported from Premna serratifolia by earlier
researchers. Preliminary screening of phytochemical compounds in Premna serratifolia
revealed the presence of sterols and triterpenes (Debelmas et al., 1973), resin, alkaloids
[premnine (Basu and Dandiya, 1947), ganikarine (Basu and Joneja, 1949), premnazole
(Barik et al.,1992) and ganiarine], Flavonoid-luteolin (Dasgupta et al., 1984),
glycosides, tannins, phenolic compounds, carbohydrates, amino acids and some
36

unsaturated aromatic hydrocarbons (Chopra and Chopra, 1955; Chopra et al., 1956;
Nadkarni, 2007; Mali and Bhadane, 2010). According to Kartick (1984), this drug plant
contains a volatile alkaloid to the extent of 0.05% and some resins soluble in alcohol.
The alkaloid premnine decreases the force of contraction of heart and produces dilation
of the pupils (Dey, 1980). Yuasa et al. (1993) isolated a new phenylethanoid, and other
phenolic compounds from its stem. As reported by Rastogi and Mehrotra (1990), a
compound with mp 155 was isolated from root bark of Premna integrifolia and it was
active against gram-positive organisms. Rastogi and Mehrotra (1991) also reported sitosterol isolated from the leaves and stem bark and betulin from the stem bark of
Premna integrifolia. From the leaves, a verbascoside-iridoid glycoside conjugate was
isolated along with premnafolioside (Otsuka et al., 1993). According to Chunekar
(2005), this plant mainly contains p-methoxy cinnamic acid and linalol, linoleic acid, sitosterol and flavone luteolin. As reported by Rajendran et al. (2008) and Rao et al.
(1985), iridoid glycoside is one of the major active principles of Premna serratifolia. In
an attempt to standardize Dasamula containing formulations, Alam et al. (1993)
reported large quantities of alkaloids from Premna serratifolia. Researchers like Ky et
al. (2005), Caldecott and Tierra (2006), Daniel (2006), Bagchi et al. (2008) and Hang et
al. (2008) also reported the presence of premnine, ganiarine,ganikarine, premnazole,
aphelandrine, pentacyclic terpene betulin, caryophellene, premnenol, premnaspirodiene
and clerodendrin-A from this medicinal plant. The aqueous extracts of this plant
showed a powerful action on the uterus and gout of the experimental animals (Khare,
2007). According to this study, the leaves contain an isoxazole premnazole, which was
found to reduce granuloma formation in rats (34.62%) and its activity was comparable
to phenylbutazone (35-36%). The study suggested that premnazole acts probably by
controlling the activity of the adreno-corticatropic hormone.
Phytochemical methods adopted for the isolation and characterization of novel
compounds
Earlier researchers have adopted different methods to identify, screen and isolate
37

phytochemical compounds from the roots, stem and leaves of Premna serratifolia. The
preliminary phytochemical screening followed by evaluation of biological properties
were reported by researchers like Rajendran et al. (2008), Rajendran et al. (2009),
Rajendran
et al. (2009), Rajendran (2010), Rajendran and Krishnakumar (2010),
Rajendran and Saleem (2010), Thirumalai et al. (2011), Kumar et al.(2013a, 2013b) and
Muthukumaran et al. (2013). Some researchers have adopted successive extraction of
the plant materials in different organic solvents like petroleum ether/ hexane,
chloroform, ethyl acetate, methanol/ ethanol and water or combination of different
solvent systems for the isolation of different classes of compounds (Mali and Bhadane,
2010; Rajendran, 2010). Isolation of major compounds (alkaloids, flavonoid glycosides,
steroids) by TLC/ HPTLC profiling were also reported (Gokani et al., 2008; Rajendran
et al., 2008; Gokani and Shah, 2009; Yadav et al., 2011; Yadav and Gupta, 2013).
As a part of developing chemical markers for ensuring the quality standards in
Ayurvedic formulations, Yadav et al. (2010) isolated three novel diterpenoids from the
root bark of Premna integrifolia and their structure were identified from their 1D and
2D NMR data. Further, these diterpenoids were also evaluated for antibacterial activity
(Yadav et al., 2010). A sensitive, selective and robust densitometric High Performance
Thin Layer Chromatographic method was developed and validated for the
determination of diterpenoid compounds. Diterpenoids 1,3,8-trihydroxy-pimara-15ene(A),
6,11,12,16-tetrahydroxy-7-oxo-abieta-8,11,13-triene
(B)
and
2,19-
dihydroxy-pimara-7,15-diene (C) were used as chemical markers for the standardization

of Premna integrifolia plant extracts. The separation was performed on silica gel 60F
(254) High Performance Thin Layer Chromatography plates using hexane: acetone:
ethylacetate (60:20:20v/v) as mobile phase. The quantification of diterpenoids was
carried out using densitometric reflection/ absorption mode at 475nm after postchromatographic derivatization using vanillin-sulfuric acid reagent. A precise and
accurate quantification was performed for compounds A, B and C in the linear working
concentration range of 1-10g/spot with good correlations (r(2)=0.9985, 0.9996 and
38

0.9992, respectively). The method was validated for peak purity, precision, robustness,
limit of detection (LOD) and quantification (LOQ) as per the International Conference
on Harmonization (ICH) guidelines. Specificity of quantification was confirmed using
retention factor (Rf) and spectra correlation of markers in standard and sample tracks
(Yadav et al., 2011).
Yadav et al. (2013) also reported two new furofuran lignans premnadimer (1) and
40-hydroxyasarinin 10 glucopyranoside (2) along with 9 known compounds isolated
from the stem bark of Premna integrifolia. Their chemical structures were elucidated
using detailed spectroscopic studies. Their relative configurations were established
using analysis of NOESY correlations and coupling constants observed in 1H NMR.
Compounds 1 and 2 together with four known iridoid glycosides were evaluated for
radical scavenging and ferric reducing antioxidant power. Radical scavenging activity
was
found
maximum
in 4-hdroxy-E-globularinin
followed
by 19-0-trans-p-
coumaroylcatalpol and the new dimer. In FRAP assay, premnosidic acid, 10-0-trans-pcoumaroy1-6-0--9-rhamnopyranosyl catalpol showed maximum ferric reducing ability
supported by high reducing power.
As a continuation of their previous studies, Yadav and Gupta (2013), attempted to
develop a High Performance Thin Layer Chromatography (HPTLC) method for
quantitative estimation of iridoid glycosides [10-O-trans-p-coumaroylcatalpol; 4hydroxy-E-globularinin; and premnosidic acid] from the stem bark of Premna
integrifolia. Separation was performed on silica gel 60 F254 HPTLC plates. The solvent
system consisted of ethyl acetate- methanol- H2O- acetic acid (80:12:6:2 v/v).
Densitometric analysis of iridoids was carried out in the absorbance mode at 510 nm
after post-chromatographic derivatization using vanillin-sulphuric acid reagent. The
method was validated as per the International Conference on Harmonization (ICH)
guidelines. This HPTLC method was found to be reproducible, accurate, and can detect
iridoids at a nanogram level. Hence, this HPTLC method can be employed as an
important tool in the quality control method for polyherbal formulations.
39

Gokani and Shah (2009) isolated and quantified a chemical marker compound viz.,
Clerodendrin-A from the roots of Clerodendrum phlomidis by simple and precise
method of HPTLC using n-hexane: ethyl formate (7:3) as mobile phase, pre-coated
TLC plates (silica gel 60 F254) as stationary phase and H2SO4 as derivatizing agent.
The method was validated in terms of linearity, precision, repeatability and accuracy.
The limit of detection and limit of quantification was determined on the basis of signal
to noise ratio. Content of clerodendrin-A was found to be 0.073 and 0.04% w/w in
Clerodendrum phlomidis and Premna integrifolia respectively.
Different methods viz., steam distillation, vacuum distillation from a hexane
concentrate and fractionation by silica gel chromatography, GC and GC/MS were
employed for the analysis and quanification of volatile compounds. Teai et al. (1998)
identified ninety-four compounds from Premna serratifolia representing about 81% of
the distillate. The major components identified are: 1-octen-3-ol (16.9%), (Z)-3-hexenol
(10.2%),
2-phenylethylalcohol (8.9%),
(E,
Z)-2,4-nonadienal(6.2%),
(E,Z)-2,6-
nonadienal (5.0%) and linalool (4.4%). In another study Rahman et al. (2011) identified
twenty-nine compounds representing 94.81% of the total leaves oil from Premna
serratifolia. The major compound identified are: Cyclohexane (1.03%), Hexan-1-ol
(0.65%), -Pinene (0.86 %),1-Octen-3-ol (8.21%), -Pinene (1.11%), 3-Octanol
(0.97%), 1,8-Cineole (0.93%), cis-2-Octenal (1.13%), Phenylethyl alcohol (5.81%),
Indole (0.91%), Decanal (0.87%), Dodecane (0.49%), Damascenone (0.18 %), Eugenol
(6.69%),
Azulene
(0.61%),
Isoeugenol
(1.83%),
-Caryophyllene
(0.92%),
Benzofuranone (0.98%), -Humulene (14.21%), Spathulenol (12.12%), Caryophyllene

oxide (2.60%), Cubenol (1.67%), Tetradecanal (0.32%), Tumerone (0.83%),
Pentadecanol (0.64%), Pentadecanoic acid (0.23%), Hexadecanoic acid (1.06%),
Eicosane (0.62%) and Phytol (27.25%).
Singh et al. (2011)b also made an attempt to study the chemical constituents of the
leaves and roots of Premna serratifolia. The alcoholic extract of Premna serratifolia
leaves were analysed and reported many compounds from ethanolic extract. The
40

compounds reported are: Glycerin (2.79%), 2,5-Furandione,3-methyl- (9.27%),
Benzofuran,2,3-dihydro-
(29.94%),
2-Hydroxy-3-methylbenzaldehyde
(6.39%),
Dodecanoic acid (7.88%), 2-Propenoic acid,3-(4-methoxyphenyl)- (13.84%), Phenol,4(3-hydroxy-1-propenyl)-2- methoxy- (1.54%), 2-Propenoicacid, 3-(4-methoxyphenyl),
ethyl ester (1.35%), 1,2-Benzenedicarboxylic acid,bis (2-methylpropyl) ester (2.50%),
n-Hexadecanoic acid (13.94%), Phytol (6.78%), Octadecanoic acid ethylester (1.68%)
and Octasiloxane, 1,1,3,3,5,5,7,7,9,9,11,11,13,13,15,15-hexadecamethyl (2.11%). The
alcoholic extract of Premna serratifolia roots were analysed to have Glycerin (1.14%),
2,5-Furandione,3-methyl(2.44%),
(2.89%),
Benzofuran,2,3-dihydro-
2-Furancarboxaldehyde,5-(hydroxymethyl)(9.86%),
2-Hydroxy-3-methylbenzaldehyde
(34.58%), Seychellene (2.30%), Dodecanoic acid (0.71%),1H-Cycloprop[e]azulen-7ol,decahydro- 1, 1, 7- trimethyl-4- methylene-, [1ar-(1a,4a,7,7a,7b)]- (2.98%), 2Propenoic acid, 3-(4-methoxyphenyl)- (13.99%), 2s,6s-2,6,8,8-Tetramethyltricyclo
[5.2.2.0(1,6)]undecan-2-ol (6.35%), 3,7,11,15-Tetramethyl-2-hexadecen-1-ol (1.34%),
n-Hexadecanoic acid (4.87%), Phytol (1.90%), Octadecanoic acid, ethyl ester (0.59%)
and 2-Phenanthrenol,4b,5,6,7,8,8a,9,10-octahydro-4b,8,8-trimethyl-1-(1-methylethyl)-,
(4bS-trans)- (4.77%).
Production of secondary metabolites by Tissue culture
Production of secondary metabolites by tissue culture method is a viable option for
the mass production of useful secondary metabolites produced from the roots and leaves
of medicinal plants without destroying their natural habitat. Advances in the area of cell
cultures for the production of medicinal compounds has made possible the production
of a wide variety of pharmaceuticals like alkaloids, terpenoids, steroids, saponins,
phenolics, flavonoids and aminoacids. In this respect, a study was undertaken by Singh
(2011) to evaluate the effect of leaf and root callus extract of Premna serratifolia
against selected human pathogens. The study revealed an increase in inhibitory activity
of root derived callus compared to the root and other extracts. The order of
antimicrobial activity of Premna serratifolia extracts was reported as: root callus extract
41

>natural root extract >leaf extract > natural leaf extract. In a similar investigation, Singh
and his co researchers studied the efficacy of callus extracts from the root and leaf
callus of Premna serratifolia for luteolin production and tested its anti-inflammatory
activity against carrageenan induced paw edema (Singh et al., 2012).
2.4.3. Pharmacological studies on Premna serratifolia
Today, plant derived drugs have paramount importance for the treatment and
prevention of diseases. Worldwide, there has been an upsurge of interests in adopting
and studying traditional systems and exploring their potential applications in developing
novel healthcare systems. In the traditional system Premna serratifolia is used for
Vatavyadhi and Amavatham (rheumatism and arthritis), nervine/ neuralgic complaints;
urinary complaints, prameha (diabetes), digestive disorders (appetizer, deranged
digestion, dyspepsia, diarrhea, and also as laxative), liver diseases, cardiac disorders,
chest pain, obesity, piles, glandular enlargement, muscular pain (body pain, head ache,
back ache), fever, skin diseases, gonorrhea, asthma, respiratory problems [acting as
kaphagna removing obstruction of respiratory tract] (George, 2006). Many
pharmacological studies have been undertaken during the last decade to evaluate the
indigenous biological properties of Premna serratifolia as described in traditional
manuscripts and folklore literature. A brief review of the works reported with special
reference to major biological activities is presented below.
Antiarthritic and anti-inflammatory activity

Rheumatic disease is one of the oldest diseases of mankind affecting the people
in the active period of their life and no substantial progress has been made in achieving
a permanent cure for this disease. The inflammatory process is a series of events that
can be elicited by numerous stimuli such as infectious agents, ischaemia, antigenantibody interactions and thermal or physical injury through years of ingenious
synthesis and structural modifications. The present review on antiarthritic and antiinflammatory property of Premna serratifolia is attempted in the above context. Few
42

studies were reported to validate the ethno-pharmacological claim regarding the antiarthritic and anti-inflammatory properties of Premna serratifolia. Karthikeyan and
Deepa (2010b, 2011) evaluated the acute toxicity and anti-arthritic activity of Premna
corymbosa (syn. Premna serratifolia) ethanolic leaf extract (PCEE) in experimental
animals. In the acute toxicity study, a single dose of PCEE, 2000 mg kg (-1) body
weight, was administered. The animals observed for 48h showed no clinical signs, no
mortality and the extract was found to be safe. To evaluate the anti-arthritic activity,
PCEE in Complete Freund's Adjuvant (CFA)- induced arthritis in rats were conducted.
The results indicated that the long-term treatment significantly (p<0.01) suppressed the
development of chronic arthritis induced by CFA. The study thus established the antiarthritic activity of Premna corymbosa (Premna serratifolia) leaves. In a similar study,
Rajendran and Krishnakumar (2010) monitored the anti-arthritic activity of ethanolic
extract of the wood of Premna serratifolia by Freund's adjuvant induced arthritis model
in albino rats. The results of the investigation revealed significant anti-arthritic activity
against adjuvant induced arthritis thereby justifying its therapeutic role in arthritic
condition. Rathore et al. (1977) reported the anti-inflammatory property of Premna
serratifolia. Jantan et al. (1996) reported significant PAF receptor binding activity for
ethanolic extract of Premna integrifolia when Malaysian medicinal plants were
subjected to their inhibitory activity. This finding provides additional evidence for the
anti-inflammatory property of Premna serratifolia. In another study, Kumari et al.
(2011) conducted a comparative experimental evaluation of anti-inflammatory activity
of the leaves of Premna obtusifolia L. and Premna latifolia Roxb. in Charles foster rats.
As revealed in this study, both species of Premna are having significant antiinflammatory activity and among the two species, Premna latifolia was found to have
better activity. According to Barik et al. (1992), premnazole, an isoxazole alkaloid
isolated from Premna integrifolia L. and Gmelina arborea L. (Verbenaceae)
demonstrated significant anti-inflammatory activity in reducing cotton pellet-induced
granuloma formation in rats. As reported by Barik et al. (1992), the anti-inflammatory
activity was comparable to that of phenylbutazone. Studies conducted by Singh et al.
43

(2012) revealed better luteolin production and good anti-inflammatory activity of the
root callus extract of Premna serratifolia than its normal root extract against
carrageenan induced paw edema.
The anti-inflammatory and antioxidant activities of Premna integrifolia was also
studied by Gokani et al. (2011). As revealed in this study, pre-treatment with a single
dose of methanolic extract of Premna integrifolia (PIM) (300 mg/kg b.w.) produced
significant inhibition on carrageenan-induced rat hind paw edema, histamine induced
wheal formation and acetic acid-induced mouse vascular permeation. In a 7-day study,
daily administration of PIM suppressed formalin induced paw edema and cotton pelletinduced rat granuloma formation. The extract also showed significant inhibition of
cyclo-oxygenase (COX-I) activity on rat uterus and plasma membrane stabilization. The
study also revealed the antioxidant activity (in vitro) of the plant extract in terms of anti
radical, superoxide scavenging, erythrocyte membrane stability, anti lipid peroxidation,
hydroxyl radical scavenging, nitric oxide scavenging and reducing power (ferric
thiocynate method and -carotene bleaching test) assays. The results demonstrated the
anti-inflammatory activity of Premna integrifolia roots in various experimental models
through their antihistaminic, antikinin, COX-inhibitory and antioxidant action, justifing
its traditional use.
Antimicrobial activity
The root bark (Kurup, 1964; Kapoor, 2001), essential oil (Rahman et al., 2011),
stem wood and stem bark (Rajendran, 2010; Rajendran and Saleem, 2010) of Premna
seratifolia were subjected to antimicrobial screening.
The antimicrobial activity of the alcoholic extract of the root bark from fresh
roots of Premna integrifolia was studied against gram-positive organisms by Kurup
(1964). The anti-bactericidal activity of the extract (in [mu] g/cm3) were noted in:
Staphylococcus aureus 0.3; Bacillus subtilis [long dash] 0.3 and Streptococcus
haemolyticus [long dash] 0.25. However, the extract was not active against E.coli,
44

Salmonella typhosa and Shigella dysenteriae. Kapoor (2001) investigated the
antibacterial activity of the phenolic substances from the root-bark of Premna
serratifolia against Staphylococcus aureus, Bacillus subtilis and Streptococcus
haemolytics. In another investigation, Rahman et al. (2011) examined the chemical
composition of the essential oil of Premna integrifolia L. and tested the efficacy of the
oil and various organic extracts as an antibacterial potential. The chemical compositions
of the essential oil were analyzed by GC-MS. Twenty-nine compounds representing
94.81% of the total leaves oil were identified. The oil (15 L disk1) and extracts (300
g disk1) of Premna integrifolia displayed a great potential of antibacterial activity
against Sarcina lutea (IFO 3232), Bacillus subtilis (IFO 3026), Escherichia coli (IFO
3007), Pseudomonas sp. (ATCC 13867), Klebsiella pneumoniae (ATCC 10031) and
Xanthomonas campestries (IAM 1671) with their respective zones of inhibition of 12.0
1.2 to 22.1 1.2 mm and MIC values of 62.5-250 g mL1. Rajendran (2010) studied
the antimicrobial activity of stem wood and bark of Premna serratifolia against bacteria
(Staphylococcus, Escherichia coli, Klebsiella pneumonia, Salmonella typhi, Salmonella
paratyphi A, Salmonella paratyphi B, Pseudomonas aeruginosa, Vibrio cholera) and
fungus (Aspergillus flavus, Aspergillus niger, Penicillium notatum, Candida albicans).
As revealed in this study, ethyl acetate, ethanol and aqueous extracts exhibited
significant antibacterial activity; n-hexane and chloroform extracts exhibited moderate
antibacterial activity and these five extracts exhibited significant antifungal activity.
Premna serratifolia L., was also screened to evaluate in vitro antimicrobial activity
against the selected human pathogenic organisms including gram +ve and gram -ve
bacterial organisms. The study suggests that Premna serratifolia L. is a promising
medicinal plant for the treatment of various pathogenic diseases which in future can be
developed as a potential antimicrobial agent with reduced toxicity and adverse effects
when compared with the synthetic chemotherapeutic agents (Rajendran and Saleem,
2010). In another study, Singh (2011), evaluated the antimicrobial activity of natural
leaves, roots of Premna serratifolia L. and its respective callus induced with help of
various plant growth regulators against the selected human pathogens (Bacillus sp.,
45

Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Non-haemolytic
Streptococci, Streptococcus epidermidis, Pseudomonas aeruginosa, Salmonella
typhimurium). As reported in this study, increased inhibitory activities of callus extracts
were found better than the natural plant material extracts (Singh, 2011). The results of
the above mentioned studies suggest that the natural products derived from the roots,
leaves and callus of Premna serratifolia can be used as potential preservatives or
antimicrobial agents in food, pharmaceutical and/or agro industries.
Antidiabetic activity
To date, over 250 million people worldwide are known to suffer from diabetes.
The disease is now reported to be increasing in emerging and many developing

countries at epidemic proportions. India is among the highest affected emerging
economy country with 41 million diabetes cases reported in 2006 and a projected rise to
70 million by the year 2025 (WHO, 2012). In the above context, the pharmacological
studies on antidiabetic property of Agnimantha remain relevant. Studies conducted by
Kar et al. (2003) revealed the hypoglycaemic activity of Premna integrifolia along with
24 known medicinal plants. Dash et al. (2005), studied the antihyperglycemic activity
of the roots of Premna corymbosa which was conducted on both normoglycemic and
hyperglycemic rats at dose levels 200-400mg/kg. As revealed in this study, the extract
produced marked reduction in blood glucose concentration at tested dose levels in a
dose dependent manner. However in normoglycemic animals the extract at 400mg/kg
dose level produced significant reduction of blood glucose at the 8th hour of
administration. Thiruvenkata and Jayakar (2010) studied the effect of extracts of
Premna corymbosa on blood glucose levels and serum lipid profile (total cholesterol,
triglycerides, phospholipids, low density, very low density and high density lipoprotein)
in the diabetic and non diabetic rats. The study revealed significant reduction in Total
cholesterol, LDL cholesterol, VLDL cholesterol and improvement in HDL cholesterol
in diabetic rats, there by proving the antidiabetic and antihyperlipidaemic activity of this
medicinal plant in HDL cholesterol in diabetic rats (Thiruvenkata and Jayakar, 2010).
46

Majumder et al. (2014), investigated antidiabetic activity along with in vitro
antioxidant activity of the methanolic bark extract of Premna integrifolia (MEPI). Oral
glucose tolerance test (OGTT), normoglycemic test and alloxan induced diabetic test
were conducted for the evaluation of antidiabetic activity. In addition, total phenolic
content, total antioxidant activity, scavenging of 1,1-dipheny1-2-picrylhydrazyl (DPPH)
radical as well as reducing power assessment were used to evaluate antioxidant
potentiality of MEPI. The continuous post treatment for 120 min with the MEPI
showed potential hypoglycemic activity in OGTT and normoglycemic rats. At a dose of
300 mg/kg the extract, considerable drop in elevated blood glucose level was observed
in the alloxan induced diabetic (p<0.05) rat after 7 days. The study also revealed radical
scavenging properties in the DPPH assay (IC = 8.61 0.16ps/ml)
(Majumder et al.,
2014). The results of the above mentioned studies provide additional evidences to
validate the traditional claim of the roots of Premna serratifolia for treating diabetes.
Antiobesity activity
Over the last few decades, there have been dramatic changes in the environment,
life style, life expectancy, dietary habits and behavior of people at global level. These
changes have resulted in escalating obesity rates, deteriorating the quality of life and
making shorter life expectancy in developed and developing countries of the world. In
the above context, researchers like Ghosh and Sukumar (2009), Mali et al. (2013) and
Patel and Patel (2012) have made pharmacological studies to evaluate the therapeutic
efficacy of Agnimantha as an antiobesity agent. Ghosh and Sukumar (2009) studied the
antiobesity property of Premna obtusifolia R. Br. (syn. Premna serratifolia L.) in 26
subjects with severe form of obesity and the results were compared to age and sex
matched controlled subjects. Subjects of treatment showed remarkable decrease in body
mass index (BMI), triglyceride, cholesterol-HDL ratio, uric acid, LDL-HDL ratio and
midtriceps skin-fold thickness (Ghosh and Sukumar, 2009). In another investigation
Mali et al. (2013) evaluated the anti-obesity activity of chloroform: methanol extract of
Premna integrifolia (CMPI) in mice fed with cafeteria diet. Female Swiss Albino mice
47

were divided into six groups, which received normal and cafeteria diet, standard drug
simvastatin (10 mg/kg) and CMPI (50, 100 and 200 mg/kg) daily for 40 days.
Parameters such as body weight, body mass index (BMI), Lee index of obesity (LIO),
food consumption, locomotor behavior, serum glucose, triglyceride, total cholesterol,
high density lipoprotein (HDL), low density lipoprotein (LDL), very low density
lipoprotein (VLDL), atherogenic index, organ weight and organ fat pad weight were
studied for evaluating the anti-obesity activity of Premna integrifolia. The result of this
study corroborates the result of Ghosh and Sukumar (2009). In another study, Patel and
Patel (2012) evaluated the antihyperlipidaemic activity of Premna integrifolia on
nicotine induced hyperlipidaemia in male albino rats. For the experiment, male albino
rats weighing 200-250 gm were divided into 4 groups viz. Normal control; Nicotine
control (NC); Nicotine control (NC) and Premna integrifolia (Agnimantha) (500 mg/
kg) treated; Nicotine control (NC) and Atorvastatin treated (standard control). Blood
samples were collected after 7 days, for lipid estimation. As noted in this study,
Nicotine caused significant increase in the serum Cholesterol, Triglyceride, VLDL,
LDL & significant reduction in HDL level. Premna integrifolia and Atorvastatin
treatment showed significant prevention in increased serum Cholesterol, Triglyceride,
LDL as compared to Nicotine control (NC) group. While HDL level was significantly
increased in treated and standard group as compared to Nicotine control (NC) group.
Cardioprotective activity
Heart failure is an extremely severe cardiac state associated with high mortality
rate. In traditional system of medicine, Premna serratifolia is considered as an

important plant having the potential to tone up the functioning of heart and circulation
of blood. Rajendran and Saleem (2008) tested the cardioprotective effect of ethanol
extract of Premna serratifolia Linn. on Isoproterenol administered experimental
myocardial infarction in rats. The cardioprotective effect of ethanol extract of Premna
serratifolia L. on Isoproterenol induced myocardial infarction in rats was confirmed by
ECG study, electrophoresis analysis of serum protein, serum A/G ratio, biochemical
48

studies such as heart tissue proteins, glycogen, nucleic acids and blood glucose. The
ethanol and aqueous extracts of stem bark and stem wood of Premna serratifolia were
subjected to screening for their cardiac stimulant activity using isolated frog heart
perfusion technique and biochemical parameters in heart tissue and serum of albino rats
after administrating the extracts for 7 days (Rajendran et al., 2008). The preliminary
phytochemical detection and HPTLC profiling were monitored along. The ethanol
extract produced significant positive ionotropic and negative chronotropic actions
similar to that of digoxin on frog heart while aqueous extract produced positive
ionotropic and chronotropic effects similar to that of adrenalines. A significant decrease
in membrane Na+ K+ ATPase and Mg2+ ATP ase and an increase in Ca2+ATP ase
further confirmed the cardiotonic activity. The results suggest that the ethanol extract
produced cardiotonic effect and the aqueous extract produced -adrenergic effect.
Ethanol extract showed positive reaction for alkaloids, glycosides, flavonoids and
steroids whereas the aqueous extract showed positive reaction for alkaloids, glycosides
and phenolic compounds. HPTLC profile of ethanol extract showed 10 peaks in the
solvent system of n-hexane: ethylacetate (3:1) at 260 nm and aqueous extract showed 7
peaks in the solvent system of chloroform: methanol: water (7:2.6:0.4) at 260 nm. It is
believed that phytoconstituents like iridoid glycosides, alkaloids, flavonoids and
phenolic compounds in Premna serratifolia have protective myocardial property.
Anticoagulation activity of the flavonoids in Premna integrifolia was reported in earlier
study by Gopal and Purushothaman (1984).
Gastro protective potential

Many medicinal plants and dietary nutrients have been shown to possess gastro-
protective activity. In clinical practice, peptic ulcer is one of the most prevalent
gastrointestinal disorders in developed countries. Treatments available for peptic ulcer
is generally non specific and is usually aimed at reducing the production of gastric acid
and re-enforcing gastric mucosal protection such as regular food, adequate rest and
avoidance of ulcer generating soft drinks, coffee, alcohol and tobacco. In traditional
49

medicine, the drug Agnimantha is one of the specific drugs prescribed in such
conditions to enhance digestive power and also to eliminate the ill effects of kapha and
vata. In this context, Jothi et al. (2010) studied the gastro protective potential of Premna
serratifolia L. leaves against Aspirin induced ulcer in albino rats. As revealed in this
investigation, the ethanolic extract of Premna serratifolia has significant antiulcer and
antisecretory activity when compared to the drug, Ranitidine. In a similar investigation,
Rajathi and Indumathi (2013) investigated the anticulcer activity of methanolic bark
extract of Premna serratifolia against Aspirin induced gastric ulcer models of male
Wistar rats. In Aspirin induced pylorus ligation model, various parameters viz. volume
and pH of gastric juice, total acidity, free acidity, ulcer score, ulcer index and
percentage protection were studied. Ranitidine (50 mg/kg p.o.) was used as the standard
drug. Pre-treatment with the extracts (200 & 400 mg/kg p.o.) showed significant
protection against ulcer model. In Aspirin induced pylorus ligated model, the
methanolic bark extract of Premna serratifolia showed significant decrease in the
volume of gastric juice, free and total acidity, ulcer score, ulcer index and increase in
pH of gastric juice as compared to the toxicant control group. In short, the antiulcer
assay of the leaf and bark extract of Premna serratifolia revealed a decrease in gastric
acid secretion thereby reducing the prevalence of causing peptic ulcer. Further, as
revealed in these studies, Premna serratifolia possess significant antiulcer and
cytoprotective effect. The activity may be due to the presence of phytoconstituents like
alkaloids, iridoid glycosides and flavonoids in Premna serratifolia.
Hepatoprotective activity
Liver is one of the most vital organs in our body that functions as the centre of
metabolism of nutrients such as carbohydrates, proteins, lipids and also helps in the
excretion of waste metabolites. By handling the metabolism and excretion of drugs and
other xenobiotics from the body, liver provides protection against foreign substances by
detoxifying and eliminating them. Liver disease is one of the most common causes of
mortality for both men and women and the trend for liver disease is steadily increasing
50

in developed and developing countries. The key risk factors that cause individuals to
develop liver disease are: excessive alcohol consumption, infection with hepatitis B and
C virus, obesity and related metabolic syndrome. Today, natural products and their
active principles as sources for developing novel hepatic drugs have attracted attention
since these drugs have less side effects compared to synthetic drugs.
In the above context, George (2006) evaluated the hepatoprotective activity of
the leaf and root extract of Premna serratifolia L. (500 mg/kg) by analyzing the hepatic
enzyme parameters viz., alkaline phosphatase (ALP), acid phosphatase (ACP), serum
glutamate oxalate transaminase (SGOT) and serum glutamate pyruvate transaminase
(SGPT) in the blood serum and liver of albino Wistar rats after a period of 21 days
treatment. The hepatoprotective activity was studied by comparing the levels of the
hepatic enzymes of normal group Wistar rats with that of the treatment groups (CCl4
group & CCl4 + Premna serratifolia leaf/ bark extract groups). The study revealed that
the leaves and roots of Premna serratifolia are very effective to tone up the functioning
of liver by nullifying the ill effects caused by CCl4. Besides, an attempt was made to
compare the hepatoprotective activity of Premna serratifolia with that of a proven
hepatoprotective drug plant, Eclipta alba (500mg/kg). In another study, Vadivu et al.
(2009) evaluated the hepatoprotective and in vitro cytotoxic activity of alcoholic extract
of leaves of Premna serratifolia L. Hepatoprotective activity was studied by carbon
tetrachloride induced hepatotoxicity in rats and the in vitro cytotoxic activity was
carried out by tryphane blue exclusion method using EAC cell lines. The degree of
protection in hepatoprotective activity has been measured by using biochemical
parameters such as SGOT, SGPT, ALP, bilirubin and total protein. The results
suggested that the alcoholic extract at the dose level of 250mg/kg had produced
significant (p < 0.001) hepatoprotection by decreasing the activity of serum
enzymes, bilirubin, and lipid peroxidation which was comparable to that of standard
drug Silymarin. The alcoholic extract also does exhibit the IC50 value of 75g/ml
which indicated the significant in vitro cytotoxic activity of the extract. As it is evident
from the above mentioned studies, the alcoholic extract of leaves/ roots of Premna
51

serratifolia L. is not only an effective hepatoprotective agent, but also possesses
significant antitumor activity. Muthukumaran and Pattabiraman (2010) studied the
hepatoprotective activity of the aqueous extract of Premna serratifolia L. against carbon
tetrachloride- and paracetamol- induced hepatotoxicity in rats. As revealed in this study,
Premna serratifolia exhibited significant hepatoprotective activity by reducing carbon
tetrachloride- and paracetamol- induced change in bio-chemical parameters that was
evident by enzymatic examination. The plant extract was interfered with free-radical
formation, providing hepatic protection. Acute toxicity studies revealed that the LD sub.
50 value is more than the dose of 4 g/kg body wt. (Muthukumaran and Pattabiraman,
2010).
Studies conducted by Singh et al. (2011)a using the ethanolic extract of root and
root derived callus extracts of Premna serratifolia L. against paracetamol induced
oxidative stress and hepatotoxicity in blood and liver of male albino rats provide
additional evidence to supplement the results generated by earlier researchers.
According to Singh et al. (2011)a, the effects of root callus extracts were comparable to
that of standard drug-Silymarine. Histopathological findings also suggest that the root
callus extracts of Premna serratifolia L. is effective to prevent the development of
chronic damage. It was concluded that the ethanolic extract of root callus is not only an
effective hepatoprotective agent but also possess significant antioxidant activity that can
be attributed to the flavanoid- luteolin and other alkaloids like premnine, ganikarin and
ganiarin which helps to overcome the paracetamol-induced toxicity.
Antioxidant activity
The stem bark, stem wood, roots and leaves of Premna serratifolia were
screened for their in vitro antioxidant property by researchers like Rajendran et al.
(2009)a, Selvam et al. (2010), Shilpa et al. (2012), Jain et al. (2013), Muthukumaran et
al. (2013) and Mali (2014). The stem bark and stem-wood extracted with 95% ethanol
and double distilled water were screened for their in vitro antioxidant potential
52

(Rajendran et al., 2009)a. Inhibitions of oxygen-derived free radicals, viz., assays for free
radical scavenging by DPPH, reducing power ability and nitric oxide scavenging were
performed. All the antioxidant activities were compared with standard antioxidantascorbic acid. Both the extracts of this plant showed effective free radical scavenging
activity, reducing power and nitric oxide scavenging activity. All these antioxidant
properties were concentration dependent. The highest antioxidant activity was observed
with ethanol extracts. Besides, the antioxidant activities of the ethanol extract of stembark & stem-wood were also determined by high fat diet (HFD) induced oxidative
stress in rabbits (Rajendran et al., 2009)b. The study revealed significant antioxidant
activity by lowering the enzyme levels of thiobarbituric acid reactive substance
(TBARS) and by increasing the enzyme levels of catalase (CAT), glutathione (GSH)
and super oxide dismutase (SOD), which was comparable with the standard
Atorvastatin in a dose dependent manner and also demonstrated remarkable activities to
scavenge reactive oxygen species (ROS), which may be attributed to the high amount of
hydrophilic phenolic compounds in Premna serratifolia (Rajendran et al., 2009)b. The
antioxidant activity of methanolic extract of Premna serratifolia leaf in paracetamol
intoxicated Wistar albino rats was studied by Selvam et al. (2010). The experiment
comprised of five groups such as healthy control group, disease control (Paracetamol
treated), positive control group (paracetamol + Silymarin), treatment groups (test drug +
Paracetamol) lower dose (100mg/kg b.wt.) and higher dose (200mg/kg b.wt.), having
six animals in each group. The animals were maintained in the standard laboratory
conditions and the test extract was administered p.o. The antioxidant enzymes
superoxide dismutase, catalase and glutathione peroxidase were evaluated in the blood
samples and as well as in the isolated tissue samples of liver, kidney and heart. The
disease control group showed decreased status of antioxidant enzymes in blood and
tissue samples. But the elevated levels of SOD, catalase, glutathione were observed
significantly (p<0.05, p<0.01) in liver, kidney and heart tissue samples and blood
samples of the extract administered groups. The activity was found to be dose
dependent. The overall efficacy of the extract was comparable with the standard drug
53

silymarin. The study thus confirmed the potential antioxidant activity of the methanolic
extract of Premna serratifolia in the animal model system. In another study, Shilpa et
al. (2012) investigated the in vivo antioxidant activity of ethanolic extract of Premna
corymbosa (Rottl.) root against streptozotocin induced oxidative stress in different
organs (liver, kidney, brain, heart and pancreas) of rats. Ethanolic extract of Premna
corymbosa (Rottl.) root was administered orally (200 mg/kg body weight) and the effect
of extract on enzymatic antioxidants like SOD, CAT, glutathione peroxidase (GPx),
glutathione-S-transferase (GST) and polyphenol oxidase (PPO), non enzymatic like
vitamin C, vitamin E and glutathione were determined. Lipid peroxidation like basal,
ascorbate and peroxide induced lipid peroxidation were also estimated. Glibenclamide
was used as standard reference drug. A significant increase in the activities of
superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and
reduced glutathione were observed in different organs of diabetic rats on treatment with
200 mg/kg body weight of Premna corymbosa (Rottl.) root extract and glibenclamide
for 30 days treatment. Both the treated groups showed significant decrease in lipid
peroxidation, suggesting its role in protection against lipid peroxidation induced
membrane damage.
Jain et al. (2013) studied the in vitro antioxidant activity of aqueous and
methanolic extracts of Premna integrifolia roots with special reference to DPPH,
DMSO, ABTS, Nitric oxide and Iron chelation assay. Ascorbic acid was used as the
standard. In all the models studied, the aqueous extract showed IC 50 values of 111.009,
101.369, 99.976, 109.827, 105.239g/ml and the methanolic extract showed IC 50
values of 98.252, 96.559, 88.163, 100.631, 95.005g/ml for DPPH, DMSO, ABTS,
Nitric oxide and Iron chelation assay, respectively.
Muthukumaran et al. (2013) in another study, investigated the antioxidant effect
of the wood extract Premna serratifolia by various antioxidant assays (1,1-diphenyl-2picrylhydrazyl (DPPH), 2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS)
and hydrogen peroxide scavenging method). The antioxidant activities were compared
54

to standard antioxidantascorbic acid. The wood extract showed a significant
antioxidant activity in DPPH, ABTS and H2O2 scavenging methods. In a recent
investigation, Mali (2014) evaluated the beneficial effect of extracts of Premna
integrifolia root on human leucocytes and erythrocytes against hydrogen peroxide
(H2O2) induced oxidative damage. Chloroform: methanol (1:1) extract of Premna
integrifolia (CMEPI) and aqueous extract of Premna integrifolia roots were used to
screen CAT, SOD, GPx, GSH and LPO levels in H2O2 induced oxidative damage. The
study revealed that, there was an increase in the CAT, SOD, GPx and reduction of the
GSH and LPO levels in H2O2 group compared with the control. Premna integrifolia
root extract treated groups showed the reduction of CAT, SOD, GPx and increased in
the GSH and LPO levels as compared with H2O2 group. CMEPI was found to be more
effective than aqueous. The study suggests that, extracts of Premna integrifolia root
possess beneficial effect on human leucocytes and erythrocytes against H2O2 induced
oxidative damage, which has substantiated their use in ethnomedicine as an antioxidant.
Observed effect was attributed due to the flavonoid and phenol contents in the plant.
The results obtained from the above mentioned studies indicated that entire parts
viz., leaves, stem and roots of Premna serratifolia are potential source of natural
antioxidants. The in vitro antioxidant activity of this plant shows the presence of antioxidant principles present in its roots, stem and leaves. These findings points to the
great interest of the Premna serratifolia whose phytochemistry and phytopharmacology
should be subjected to detailed investigation so as to bring forth all possible
phytotherapeutic uses of the species, Premna serratifolia and its related species in
different parts of the world.
Immunomodulatory, Antinociceptive, Analgesic, Neuropharmacological,

Anticonvulsant, Tumor cell suppression and Anti-parasitic activities
Studies on various pharmacological aspects such as immunomodulatory,
antinociceptive,
analgesic,
neuropharmacological,
anticonvulsant,
tumor
cell
55

suppression and anti-parasitic activities of Premna serratifolia L. were subjected to
detailed investigations by researchers in recent years. The results emerged from these
studies are summarized below.
Immunomodulatory potential: Immunomodulatory potential of Agnimantha
drugs with special reference to the roots Clerodendrum phlomidis and Premna
integrifolia were investigated by Gokani et al. (2007). Methanol root extracts of the
above mentioned plants (300 mg kg-1 x 7 days) were administered orally to mice prior
to immunization with Sheep Red Blood Cells (SRBC) and it resulted in a significant
increase in haemagglutinating antibody titre, plaque forming cell assay and delayed type
hypersensitivity to SRBC. Clerodendrum phlomidis and Premna integrifolia enhanced
the non specific immune response in carbon clearance test and showed significant
immunoprophylactic effect, when tested on E. coli induced abdominal sepsis. In the
study Clerodendrum phlomidis showed higher response to specific immune activity as
compared to Premna integrifolia, where as in case of non specific immune activity both
the roots showed almost equal response.
Antinociceptive activity: Karthikeyan and Deepa (2010)a evaluated the acute
toxicity and antinociceptive activity of an ethanolic extract of Premna corymbosa in
animal models. In the acute toxicity study, the ethanolic extract showed no clinical
signs and mortality of the animals and was found to be safe. In the acetic acid-induced
writhing model, the ethanolic extract at a dose of 200 or 400 mg kg (-1) body weight
significantly (p<0.01) inhibited the writhing response by 42.57% and 54.67%,
respectively. In the hot plate test, the extract produced a significant (p<0.01) increase in
latency with 34.50% and 51.08% of protection in a dose-related manner. The study
established the analgesic properties of Premna corymbosa. It is speculated that sitosterol or luteolin in the ethanolic extract might have contributed to the
antinociceptive activity of Premna corymbosa.
56

Analgesic,
Neuropharmacological
and
Anticonvulsant
property:
In
traditional medicine, Premna serratifolia is recommended as an analgesic to relieve

body pain, back ache and head ache. In this back drop, Karmakar et al. (2011) assessed
analgesic activity of the ethanolic extract of the leaves of Premna integrifolia using
acetic acid induced writhing model in mice. In analgesic activity, the extract produced
52.17% (p<0.01) acetic acid induced writhing inhibition in mice at the dose of 500
mg/kg body weight, which was comparable to diclofenac sodium 65.21% (p<0.01) at
the dose of 25 mg/kg body weight.
In a recent investigation, Khatun et al. (2014) studied neuropharmacological,
analgesic, and anti-inflammatory activities of the methanolic bark extract of Premna
integrifolia (MEPI). Neuropharmacology study was done by open field and hole-cross
test whereas acetic acid writhing test and formalin induced pain was done for analgesic
activity of MEPI. Carrageenan induced inflammatory model was considered for antiinflammatory activity evaluation. A statistically significant (p<0.05) decrease in
locomotor activity was observed at all doses in the open-field and hole-cross tests. The
extract significantly (p<0.05) and dose dependently reduced the writhing reflex in the
acetic acid-induced writhing test as well as licking response in the formalin induced
inflammatory pain. At 200 mg/kg body weight dose, MEPI showed 71.16% inhibition
in carrageenan induced anti-inflammatory activity.
Baby et al. (2011) evaluated the anticonvulsant effect of the extract of Premna
corymbosa against Pentylenetetrazole and maximal electroshock induced convulsions in
mice.
Tumor cell suppression potential: Selvam et al. (2012) evaluated the free
radical scavenging activity and tumor cell suppression potential of the methanolic leaf
extract of Premna serratifolia in various in vitro model systems. The superoxide radical
scavenging activity, nitric oxide radical, hydroxyl radical, DPPH radical and ABTS
radical scavenging activity and lipid peroxidation were determined. The tumor cell
57

suppression cell potential was determined in three different cancer cell lines MCF7
(breast cancer), HepG2 (liver cancer) and A549 (lung cancer) by SRB assay. As noted
in this study, the methanolic extract of Premna serratifolia has free radical scavenging
activity against superoxide radical, nitric oxide radical, hydroxyl radical, DPPH radical,
ABTS radical and also has the potential to inhibit lipid peroxidation. The efficacy was
dose dependent as it is very clear from the IC 50 value. The test extract showed
cytotoxic activity against MCF7, HepG2 and A549 cell lines. The GI50, TGI and LC50
values were determined against each cell line and compared with standard drug
Adriamycin. Thus the study proved the free radical scavenging activity and tumor cell
suppression potential of Premna serratifolia leaf in the selective in vitro model systems.
Anti-parasitic activity: Desrivot et al. (2007) reported antiparasitic activity for
Premna serratifolia against Leishmania donovanim with IC 50 values between 0.55g/ml.
In short, the pharmacological evaluation studies on Premna serratifolia L. so far
reported with special reference to antiarthritic, anti-inflammatory, antimicrobial,
antidiabetic, antiobesity, cardioprotective, gastro protective, hepatoprotective and
antioxidant activities have shown promising results for bioprospecting Premna
serratifolia L. as a Neutraceutical Panacea.
2.5. Conclusion
The taxonomic review of the genus Premna and the species Premna serratifolia
L. in particular revealed much perplexity in delimiting the different species of the genus
Premna with special reference to Premna integrifolia, Premna serratifolia, Premna
corymbosa and Premna obtusifolia and also with respect to different morpho-variants of
Premna serratifolia. Even though many studies have been undertaken by earlier
researchers based on the morphology and taxonomy of related species of Premna
serratifolia, the confusion was not fully resolved due to high polymorphism among
different morphotypes of this species. In the Ayurvedic/ traditional systems of medicine,
58

Premna serratifolia has a significant position. However, this plant has a controversial
drug status in Ayurveda since the source drug Agnimantha is equated to many species
like Clerodendrum phlomidis L., Clerodendrum inerme (L.) Gaertn., Premna
serratifolia L., Premna latifolia Roxb. and Premna obtusifolia L. in different parts of
India. The major source of the drug in Eastern and Central parts of India is Premna
obtusifolia and that in the North and Western regions is Premna latifolia. Clerodendrum
phlomidis is considered as the source plant in certain North Indian states and in Gujarat
state while Clerodendrum inerme is the accepted source plant of Kshudragnimantha in
certain parts of South Indian states especially Kerala state. This controversial drug
status of Premna serratifolia with other substitute medicinal plant species viz.,
Clerodendrum phlomidis and Clerodendrum inerme necessitates further studies based
on phytochemical and pharmacognostic characteristics of these plant species. The
earlier pharmacognostic and phytochemical studies reported were based on either
limited samples or morphotypes of Premna serratifolia. A perusal of literature revealed
that no research studies have so far been done based on the comparison of
pharmacognostic and phytochemical characteristics of the different morphotypes of
Premna serratifolia L. in Kerala. The present investigation is an attempt in this context
and it aims to study and compare the different morphotypes of Premna serratifolia in
Kerala using relevant tools and techniques in Phytochemistry and Pharmacognosy. It is
expected that the results emerged from this study will be helpful to resolve the
controversial status of Premna serratifolia in taxonomy and Ayurvedic system of
medicine.
59

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REVIEW OF LITERATURE

2.1. GENUS PREMNA: AN OVERVIEW

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Balarishtam, Premna serratifolia is used as a major constituent. The major rasayanams

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powdered roots of Premna

Indukanthakwatham, Dhanwantharakwatham, Varanadikwatham, Sapthasarakwatham,

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Harimantha, Havirmantha, Jaya, Jayanthi, Jayee, Jyotishka, Kanika, Karnika, Kethu,

: Arni, Agetha, Arani, Ganiari

: Munnai, Munnay, Munney, Munni, Munni-vayz, Pasumunnai,

: Gabbunelli, Ghebunelli, Kanika, Karnika, Nagura, Nelli,

Malayalam (Kerala) : Munja, Munna, Appa, Appel, Kozhychedy.

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integrifolia and Clerodendrum phlomidis respectively by many scholars. While writing

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et al. (2009), Rajendran (2010), Rajendran and Krishnakumar (2010),

dihydroxy-pimara-7,15-diene (C) were used as chemical markers for the standardization

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Benzofuranone (0.98%), -Humulene (14.21%), Spathulenol (12.12%), Caryophyllene

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Antiarthritic and anti-inflammatory activity

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The disease is now reported to be increasing in emerging and many developing

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rate. In traditional system of medicine, Premna serratifolia is considered as an

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Gastro protective potential

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