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Acorus Calamus L.: An Insight Review of Botany, Chemistry, Medicinal Uses and
Cultural Practice

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JCBPS; Section B; May 2016 – July 2016, Vol. 6, No. 3; 1027-1045, E- ISSN: 2249 –1929

Journal of Chemical, Biological and Physical Sciences

An International Peer Review E-3 Journal of Sciences
Available online at www.jcbsc.org
Section B: Biological Sciences

CODEN (USA): JCBPAT Research Article

Acorus Calamus L.: An Insight Review of Botany,

Chemistry, Medicinal Uses and Cultural Practice

Alok Ranjan 1*, Paras Jain 2, Binod Singh 2, Pallavi Singh 2, H.P. Sharma 2
1
Department of Botany, GPGC Obra, Sonbhadra, U.P. (India)
2
University Department of Botany, Ranchi University, Ranchi, Jharkhand (India)

Received: 07 June 2016; Revised: 23 June 2016; Accepted: 02 July 2016

Abstract: Acorus calamus is monocot plant of immense medicinal potential. The

plant grows in high water location, especially on damp soil. Locally known by
various names like Bachh and Sweet Flag, belongs to taxonomic Family
Acoraceae. Its ethnomedicinal uses includes the treatment of some of the dreaded
diseases like cancer, ulcer, hepatitis, spasm, schizophrenia, gout, arthritis,
anorexia etc. It is the source of myriads of phytochemicals like α- asarone, β-
asarone, VOCs etc. Due to its medicinal uses, the rhizome of Sweet Flag is in
high demand and thus over-exploited. Seed-set is very low and vegetative
propagation is the chief method of propagation. Present review covers all
possible aspects of the plant like habit, habitat, morphology, phytochemistry,
anti-microbial property, ethnomedicinal uses, cultural practices, anti-oxidant
properties. Evolutionary aspects of the plant have also been investigated. In vitro
micropropagation of plant has also been reviewed owing to the need for its
conservation.
Keywords: Acorus calamus, Medicinal plant, Ethnobotany, Biological activity,
Phytochemicals.

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Acorus Calamus L … Ranjan et al.

INTRODUCTION

Mother earth has bestowed to the mankind and various plants with healing ability for curing the
ailments of human being. This unique feature has been identified since pre historic times. The
WHO has also estimated that 80% of the world population meets their primary health care needs
through traditional medicine only. Medicinal plants are those plants possessing secondary
metabolites and are potential sources of curative drugs with the very long list of chemicals and its
curative nature. India is the eighth largest country having rich plant diversity with a total of
around 47,000 species, of which more than 7500 species are being used as medicinal plants. Plant
products are used as main source of medicine throughout the world for treating various human
ailments. About 50% of the present day medicines in the United States of America are derived
from natural sources especially from various plants 1.
The use of traditional medicine in both developing and developed countries is significantly
increasing in recent times. There is a growing demand for medicines of Ayurveda, Siddha, Unani
and Homeopathy for domestic consumption and export purposes. The world trade in plant based
drugs and its products are many fold expanding continuously; because the general awareness of
the wide spread toxicity and harmful after effects associated with the long-term use of synthetic
drugs and antibiotics.
Estimate reveals that the world trade in medicinal plants and extracts industry was growing at a
rate of 12 to 15% per annum. The export from India is to the tune of Rs 446 crores with the
present growth rate of 7%. Acorus calamus is a tall perennial wetland monocot plant from the
Araceae (Acoraceae) family. The genus Acorus derived from Acoron (coreon = the pupil of the
eye) and the species calamus is derived from the Greek word Calamos (a reed). The family
Araceae (Acoraceae) comprises about 110 genera and more than 1,800 species.
The scented leaves and rhizomes of sweet flag have been traditionally used as a medicine and the
dried and powdered rhizome has a spicy flavor and is used as a substitute for ginger, cinnamon
and nutmeg for its odor. It is known by a variety of names, including cinnamon sedge, flag root,
gladdon, myrtle flag, myrtle grass, myrtle sedge, sweet cane, sweet myrtle, sweet root, sweet rush
and sweet sedge.
A. calamus is probably indigenous to India and now found across Europe, Southern Russia,
Northern Asia Minor, China, Japan, Burma, Sri Lanka, and Northern USA. Calamus was valued
as a stimulant, bitter herb for the appetite and as an aid to the digestion. In North America, the
decoction was used for fevers, stomach cramps and colic; the rhizome was chewed for toothache
and powdered rhizome was inhaled for congestion. In Ayurvedic medicine Calamus is an
important herb, and is valued as a "rejuvenator" for the brain and nervous system, and as a
remedy for digestive disorders.
The sweet flag oil present in this plant is a unique source of oxygenated sesquiterpenes of great
structural variety 2. Apart from this terpenes a few commonly occurring steroids and xanthones
had also been reported. The rhizome of the plant has medicinal properties against bugs, moths,
lice, emetic stomach in dyspepsia; etc 3. Common names of Acorus calamus which are used in
different parts in India are Bach (Hindi), Vashampu (Tamil), Baje (Kannada) and Vasa (Telugu)

1028 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

TAXONOMY
Kingdom: Plantae
Division: Magnoliophyta
Class: Liliopsida
Order: Acorales
Family: Acoraceae
Genus: Acorus
Species: calamus/ A. aromaticus / A. calamus var. americanus

VERNACULAR NAMES

English- Sweet Flag

Ayurvedic- Vacha
Unani- Bacch
Hindi- Bajai, Gora-bach, Vasa Bach
Marathi- Vekhand
Tamil- Vashambu
Telugu- Vadaja, Vasa
Kannada- Baje
Malayalam- Vayambu
Sanskrit- Bhutanashini, Jatila

BOTANY

A. calamus is a perennial plant with creeping habit. It shows profuse branching. Rhizome of the
plant is aromatic. It is cylindrical and up to 2.5 cm thick, purplish-brown to light brown in colour
externally and white internally. The leaves of A. calamus L. has a single prominent midrib.
Secondary veins on both sides are slightly raised and many. Tertiary veins are very fine. The
leaves are between 0.7 and 1.7 cm wide, with average of 1 cm. The sympodial leaf of A. calamus
is somewhat shorter than the vegetative leaves.
The margin is curly edged or undulate. Flowering or fruit set in Acorus is very rare, but when
they do, the flowers are 3 to 8 cm long, cylindrical in shape, greenish brown and arranged in a
multitude of rounded spadix inflorescence. The spadix can reach a length between 4.9 cm to 8.9
cm at the time of expansion. The fruits are small and berry-like, containing few seeds. Flowering
occurs from early to late summer depending on the latitude, grows wild in marshy places up to
2000 to 2200 m altitude in the Himalayas, Manipur, Naga Hills and in some parts of South India.
There are only two species in the genus Acorus-

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Acorus Calamus L … Ranjan et al.

1. Acorus calamus.
2. Acorus gramineus
Acorus gramineus is native to eastern Asia commonly called as Japanese sweet flag, Japanese
rush, grassy-leaved sweet flag, dwarf sweet flag. It is an aquatic or wetland perennial with semi
evergreen grass like foliage. It has narrow, 6 to 14 inch long (15 - 35.6 cm) glossy leaves and
looks like thick, lush grass. The leaves are carried in two ranks, like opposing fans. They are flat,
about a 0.5 inch (1.3 cm) wide and tend to flop over. Its leaf scars are brown, white and spongy. It
possesses slender roots. Its leaves are few and distichously alternate 4. The insignificant flowers,
shaped like little horns, are produced in midsummer on erect hollow stems. Usually, only plants
grown in water produce flowers.
Distribution: Acorus calamus is a native of eastern countries and also it is indigenous to the
marshes of the mountains of India. It is cultivated throughout India, ascending to an altitude of
about 2200 metres. It is also found in marshy tracts of Kashmir, Shirmaur (Himachal Pradesh),
Manipur and in Naga Hills. It is regularly cultivated in Karnataka in peninsular India 5.
Parts used: The parts used are leaves, root (rhizome) and stem. In Asia, Sweet flag has been used
for at least the last 2000 years. The ancient peoples of China used it to lessen swelling and for
constipation. In Ayurvedic medicinal practice India, the rhizomes have been used to cure several
diseases like fever, asthma and bronchitis, and as a sedative. Native tribes used it to treat a cough,
made a decoction as a carminative and as an infusion for cholic.
In Western herbal medicine the herb is chiefly employed for digestive problems such as gas,
bloating, colic, and poor digestive function. Bachh helps distended and uncomfortable stomachs
and headaches associated with weak digestion. Small amounts are thought to reduce stomach
acidity, while larger doses increase deficient acid production, a good example of how different
doses of the same herb can produce different results. It is a good sedative so that the extract is
used for epilepsy, insanity and as a tranquillizer along with valeriana jatamansi and nardostacys
grand flora. It is an ingredient of any Ayurvedic preparation “Brahmi Bati” (Budhivardhar) which
is indicated in epilepsy, coma, and hysteria and in cases of mental retardation; the same uses are
prescribed for an Acorus containing Unani drug “Ma’jun Baladur”.
CHEMISTRY OF Acorus calamus
Isolation Techniques of components in A. Calamus: A.calamus is a source of essential oil
which is responsible for the medicinal and insecticidal properties 6. The essential oil of calamus
was first isolated practically in Frankfurt in 1592 and in the dispensatorium Noricum in 1589 7.
But in recent years the isolation of various constituents from the oil has been enormously
extended by the development of the techniques of Gas- Chromatography 8. However, continuous
solvent extraction by using Soxhlete apparatus is still be the usual procedure of extraction.
Isocalamendiol 9 was isolated by steam distillation of ethereal extract which is a colourless
mobile liquid having a pleasant aromatic odor. During steam distillation of ethereal extract direct
heating on a sand-bath should be avoided to prevent charring of the fibres.
(Qudrat-I-Khuda 10). The chloroform extract residue was chromatographed over silica gel to yield
asarone, the principle compound in different isomeric forms (γ,cis and trans forms) 11 and
acoradin which were got after evaporating the benzene and petrol-ether eluate 12. Both
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Acorus Calamus L … Ranjan et al.

acrogermacrone and preisocalamendiol were successfully separated by repeated preparative TLC

using alumina [GF254 (Type-E)] in n-hexane-benzene (2:1) each in pure state 9 .The sweet flag
oil of European origin was rechromatographed on silica gel impregnated with AgNO3(15%) and
elution with hexane-EtO2 (95:5) gave calamuseone in pure state (GLC, isothermal 160°) 13. In the
isolation of Triterpenoid saponins and phenyl derivatives, the ethanolic extract was
chromatographed over silica gel column which gave compound 1 in Benzene- DCM(9:1,v/v)
fraction, compound 2 in EtOAc-Methanol (8:2,v/v) fraction and phenyl derivatives in petrol
eluate 14, 15. Galangin,a dihydroxy flavanol was isolated from the ethanolic extract residue 16.
Different parts of the plant have been subjected to exhaustive extraction and the isolated
constituents were reported in the literature, which includes Terpenoids, Steroids, Xanthones,
Lignans, Flavones and presence of traces of alkaloids.

PHYTOCHEMICALS

Phenylpropanoids: The aromatic constituents namely asarylaldehyde in roots and asarone in

leaves are responsible for the smell of volatile oil 17. Most of the phenylpropanoids were isolated
by steam distillation. An oily substance namely calamol was extracted which was found to be an
allyl trimethoxy benzene derivative. It is isomeric with asarone. Many phenylpropanoids were
extracted and isolated from chloroform extract, viz. isoeugenol methyl ether (Figure-1), γ-
asarone (1, 2, 4-trimethoxy-5 (2-propenyl) benzene) (Figure-2), Cis-asarone (cis-1,2,4-
trimethoxy-5(2-propenyl) benzene (Figure-3), Trans-asarone (trans-1,2,4-trimethoxy-5(2-
propenyl) benzene (Figure-4), Acoramone (1,2,4-trimethoxy-5 (2-propanoyl) benzene (or)
1(2,4,5-trimethoxyphenyl)-propan-2-one) Figure-(5) along with asarylaldehyde (2,4,5-
trimethoxy benzaldehyde) (Figure 6). Also phenyl indene derivatives were isolated and their
structures were derived. They were Z-3-(2, 4, 5-trimethoxy phenyl)-2-propenal (Figure-7) and 2,
3-dihydro-4,5,7-trimethoxy-1- ethyl-2- methyl-3-(2,4,5-trimethoxy phenyl) indene (Figure-8).
STRUCTURES OF PHYTOCONSTITUENTS (Figure 1-24)

1 2 3

4 5 6

1031 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

7 9 9

10 12 12

13 14 15

16 17 18

1032 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

19 20 21

22 23

24

Sesquiterpenes: Acorus calamus showed the presence of a large number of sesquiterpenes which
were extracted by steam distillation process. The fresh aerial parts yield about 0.123%, dried
rhizomes yield about 0.62 per cent and dried roots yield about 2.50 per cent on distillation 21.
Three monocyclic sesquiterpenes were isolated from rhizomes of the plant after extracting the
essential oil using n-hexane and chromatographed with pet-ether-diethyl ether fraction (10:1).
They were Shyobunone (Figure-9), Epishyobunone (Figure-10) and 2, 6-diepishyobunone
(Figure-11) 22. In the subsequent fraction a new sesquiterpene bicyclic diol namely
Isocalamendiol (Figure-12) was isolated from pet-ether-diethyl ether fraction (3:1).On rapid
elution with n-hexane afforded a mixture of Acoragermacrone (Figure-13) and

1033 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

Preisocalamendiol (Figure-14). Another two new guaiane sesquiterpenic ketones were isolated
by gradient elution using hexane-Et2O mixtures from eastern European sweet flag oil. They were
named as Calamusenone (Figure-15) and its isomer Figure-(16).
Monoterpenes: A number of monoterpenes have been reported in the plant. Some of the
monoterpenes reported are as follows α and β-pinenes, myrcene, Cymene-Para, Terpinen-alpha,
Phellandrene-beta, Terpinene-gamma, Terpinolene, Thujane (Figure-17) and Limonene (Figure-
18). They were isolated by steam distillation from volatile oil.
Xanthone glycosides: A new xanthone glycoside was isolated from rhizome part from ethanolic
extract by column chromatographed using benzene-DCM (9:1v/v) and on hydrolysis yield two
sugar moieties as D-glucose and D-galactose. It was designated as 4, 5, 8-trimethoxyxanthone-2-
O-β-D-glucopyranosyl (1 2)-O-β-D-galactopyranoside (Figure-19).
Flavones: The CHCl3 extract of the ethanolic residue afforded from CHCl3-MeOH (19:1)
eluates yellow crystalline needles. All the physical and spectral data showed the conformity with
the 5, 7-dihydroxyflavonol (Galangin) (Figure-20).
Lignans: A lignan was isolated from the rhizome part of the plant and it was designated as
acoradin (Figure-21).It was eluated using benzene from chloroform extract of the Acorus
calamus.
Steroids: β-Sitosterol (Yeh et al, 1938) is isolated along with Galangin from petrolbenzene (1:1)
eluate and forms like flaky substance. From the physical and spectral features it was confirmed to
be β- sitosterol (Figure-22).
Volatile Organic Compounds: A total of 53 VOCs have been isolated from Nepalese Acorus
calamus by SDE (simultaneous distillation extraction) method and then anlysed by GS-MS. Out
of these 53 VOCs 11 are alcohols, 14 aldehyde, 3 esters, 1 furan, 19 hydrocarbos, 4 ketones, and
1 was nitrogen containing with undefined class. Apart from that 6 unknown compounds were also
detected.

Table-1: Relative content of functional groups of vocs identified in Acorus calamus l.

S.N. Functional Groups No. of Compounds

1 Alcohol 11
2 Aldehyde 14
3 Ester 3
4 Furan 1
5 Hydrocarbon 19
6 Ketone 4
7 Miscellaneous 1
8 Unknown 6
TOTAL 59

Inorganic constituents: The oxalate and calcium content of Acorus calamus is about 2.00% and
0.078%.The oxalate is expressed as H2C2O4.

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Acorus Calamus L … Ranjan et al.

Triterpenoid saponins: Two new triterpenoid saponins have been isolated and they are
characterized as follows:
1) 1β, 2α, 3β, 19α-tetrahydroxyurs-12en-28-oicacid-28-O {-β-D-glucopyranosyl (1 2)} β- D-
galactopyranoside 18 (Figure-23)
2) 3β, 22α, 24, 29-tetrahydroxyolean-12-en-3-O-{-β-Darabinosyl (1 3)} -β-Darabinopyranoside
14
(Figure 24)

LECTINS

Lectins are present as major proteins in the storage structures of many monocotyledonous plants
and are defined as proteins possessing at least one non-catalytic domain, which bind reversibly to
a sugar moiety and are not products of immune response Lis and Sharon et al 19. Family
Acoraceae among the monocots has also been reported to be a lectin rich family Lectins are also
polyclonal activators towards human lymphocytes. Earlier reports from our laboratoryhave
revealed that lectins from the family Araceae have shown mitogenic activity Kamboj et al 20.
Report herein two novel lectins from the rhizomes of Acorus calamus (Linn.) and Acorus
gramineus (Soland in Ait.) belonging to family Araceae having significant mitogenic activity
towards human lymphocytes, BALB/c splenocytes and inhibitory activity towards murine
macrophage cancer cell lines. Two novel araceous lectins having mitogenic activity towards
BALB/c splenocytes and human lymphocytes and inhibitory potential towards murine cancer cell
lines were affinity purified and characterized from rhizomes of A. calamus (Linn.) and A.
gramineus (Solandin Ait.). Family Araceae represents a large family that, in addition to
ornamental species comprises economically important plants and Acorus plants are among these
known for their medicinal values. In India, Ayurvedic medicinal practice has used the rhizomes
of Acorus plants to cure fevers, for asthma and bronchitis, and as an all-around sedative.
Thus, apparently these lectins are similar to other reported Araceae lectins in terms of their
subunit and molecular structure. Previous reports indicated that most of the characterized Araceae
lectins such as CEA and AMA were heterotetramer composed of four polypeptides chains. Two
chains were identical but different from other two chains. Each chain had a similar size of 11–14
kDa Shangary et al 21. A lectin from family Araceae, P. ternata when searched with
BLASTP2.23 showed that this lectin has high affinity with mannose-binding lectins from C.
esculanta and A. maculatum Yao et al 22. These lectins are first synthesized as large precursor that
are post translationally processed into two polypeptides of similar size. BLAST results of Acorus
americanus have revealed (ESTs) encoding lectin protomers.
The nucleotide sequences of A. americanus when compared with the nucleotide sequences
obtained five domains of A. maculatum lectin i.e. Domain 1. bIMQGDCNLVLYQ, Domain 2.
bGWQSNTQ, Domain 3. bGELVIKQ, Domain 4. bVWSSQ and Domain 5. bGPS/A VF
revealed that these domain sequences were also 100% conserved in A. americanus. Thus,
indicating that Acorus lectins like most of the reported Araceae lectins e.g. A. maculatum that is
synthesized as large precursors containing two in tandemly arrayed mannose binding- domain
(GNA-domain). Acorus lectins were inhibited by mannose/glucose and their derivatives (data not
shown) which are epimers of each other. Acorus lectins are among the first araceous lectins
having a strong reactivity with mannose. The earlier observations on araceous lectins showed that

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Acorus Calamus L … Ranjan et al.

free mannose is a poor inhibitor of their hemagglutination activity 21. But the studies on affinity
purified araceous lectins from our laboratory have shown that these are inhibited only by
asialofetuin, a complex desialyated serum protein, and free mannose at as high a concentration as
1000 mM was unable to inhibit its activity 21. In contrast, Acorus lectins showed no reactivity
towards asialofetuin. A perusal of data on minimal inhibitory sugar concentration reveals that
most effective inhibitor for Acorus lectins is methyl-a-d-mannopyranoside which could inhibit its
activity at a concentration of 3.12 mM while free mannose was inhibitory at 6.25 mM.
Glucose or its monosaccharide derivatives were effective even at higher concentrations while N-
acetyl-d-glucosamine was as effective as mannose. In addition, both the lectins were also
inhibited by maltose and glycogen having a 1–4 glycosidic linkage. The better reactivity of
methy-a-d-mannopyranoside as compared to mannose indicates the additional importance of C1
axial methyl group in addition to C2 axial and C4 equatorial hydroxyl groups. N-acetyl-d-
glucosamine was as effective as d-mannose which may be due to C4 equatorial OH group and C2
equatorial acetamide group. In addition, A. calamus also showed reactivity towards phenyl-h-d-
glucoside, d-trehalose and dmelezitose which are glucose derivatives. Thus, the finer sugar
specificity of Acorus lectins is different from other monocot mannose-binding lectins.
Acorus lectins agglutinated rabbit erythrocytes at a minimal erythrocyte agglutinating protein
concentration (MEAPC) of 14.4 and 18.7 Ag/ml. Acorus lectins were also non-reactive towards
human ABO RBCs like other araceous lectins and monocot mannosebinding Amaryllidaceae
lectins. However, Acorus lectins could react with human ABO blood group erythrocytes after
neuraminidase treatment which indicates that sialic acid hinders cryptic receptors for these
lectins. The same behaviour was shown by ACL and AGL in the case of goat and sheep
erythrocytes. Rat and guinea pig erythrocytes were reactive with Acorus lectins. There was a 2–4
fold decrease in MEAPC after neuraminidase treatment in rabbit, rat and guinea pig erythrocytes.
Furthermore, ACL and AGL also agglutinated human lymphocytes and rat splenocytes. Like
other araceous lectins, ACL and AGL do not require metal ions for their hemagglutination
activity, as there was no effect of demetallization on hemagglutination activity.
The hemagglutination activity of these lectins declined after incubation at 55 8C and lost by 50%
at 60 8C. As reported for other araceous lectins, 25% residual activity remained even after
incubating in boiling water bath for 15 min. Denaturants, like urea, thiourea and guanidine–HCl,
also had a significant effect on their activity. Lectin activity declined to 50% at 3.0 M
concentration of urea and guanidine–HCl while in the case of thiourea, 50% decrease in the
activity was observed at 3.5 M. The decrease in lectin activity may be due to disruption of
hydrogen bonds and hydrophobic interactions. Both A. calamus and A. gramineus turned out to
be strong mitogens for BALB/c splenocytes and for human PBMC as evidenced by
lymphoproliferation after incorporation of the lectins into the cultures. Both ACL and AGL
showed significant mitogenic activity towards BALB/c splenocytes. As compared to the
unstimulated splenocytes, the ACL and AGL induced 39.7 and 28.6 fold respectively increase in
the proliferation. This response is better than that of Con A (M.I.: 20.6). Similarly, better
mitogenic response was noticed for human PBMC as compared to Con A. We titrated both the
lectins using different doses. The optimum proliferation was observed using 1.0 Ag/ml for ACL
and AGL. The proliferation response of BALB/c splenocytes by Acorus lectins was inhibited in a
concentration-dependent manner in the presence of mannose. Mannose at a final concentration of

1036 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

50 mM could cause more than 99.6% and 99.3% inhibition of mitogenic response in ACL and
AGL respectively, which indicates that lectin receptors are involved in the process of
mitogenicity. There was significant production of interleukin-2 by lectin stimulated T-cells.
Further, antibody secretion profile from the 6-day culture showed that there was no significant
production of any isotype of antibodies after incorporation of ACL and AGL but LPS-stimulated
cultures showed substantial augmentation.
Thus indicating that the proliferation observed in the cultures where lectins were added was due
to T cells and thereby establishing that both the lectins are endowed with T-cell mitogenic
response. Thus, Acorus lectins resemble best known mitogens such as PHA and Con A which are
also mitogenic for T cells. It has also been shown that PHA and Con A could prolong allograft
survival over weak histocompatibility barriers. PHA and Con A can also prevent the development
of autoimmune thyroiditis in mice. Earlier reports from our laboratory have also revealed that the
family Araceae is a rich source of mitogens. Finally, we tested whether ACL and AGL have any
inhibitory activity towards murine cancer cell lines. We employed macrophage cancer cell lines
J774 and P338D1, and T cell lymphoma A20 and WEHI-279 a B-cell lymphoma. Interestingly,
both ACL and AGL showed inhibitory activity towards J774 and WEHI-279 cancer cells. ACL
and AGL inhibited the growth of J774 to 67% and 40% respectively. Similarly, ACL and AGL
inhibited the growth of WEHI-279 to 24% and 30% respectively.
At present, it is difficult why the lectins failed to restrict the growth of P338D1 and A20 cancer
cell lines but significantly retarded the proliferation of J774 and WEHI-279 cells. This may be
due to difference in the signalling action of the lectins on different type of cancer cells. This study
can be an interesting line of investigation and these lectins can be tested for their inhibitory effect
towards various human cancer cell lines from different organs and tissues in vivo and in vitro. It
has recently shown that the CD80 and CD86 expressed on B-cell lymphoma may promote or
retard the growth by expressing anti-apoptotic and pro-apoptic molecules. Further, melatonin, a
pineal gland hormone can regulate the growth of T-cell lymphoma and normal T cell
differentially.

MEDICINAL USES

Antimicrobial activity: Phongpaichit et al 23 reported that a partially-purified fraction obtained

from column chromatographic preparation of the crude methanol extract of A. calamus rhizomes
were investigated for its antimicrobial activities on various microorganisms including bacteria,
yeasts and filamentous fungi, exhibited high activity against filamentous fungi Trichophyton
rubrum, Microsporum gypseum, and Penicillium marneffei with IC50 values of 0.2, 0.2 and 0.4
mg/ml, respectively. However, it showed moderate activity against yeasts: Candida albicans,
Cryptococcus neoformans and Saccharomyces cerevisiae (MIC 0.1 to 1 mg/ml) and low activity
against bacteria (MIC 5 - >10 mg/ml). Scanning electron microscopic observation revealed that
hyphae and conidia treated with this fraction were shrunken and collapsed, which might be due to
cell fluid leakage. Antibacterial activity of A. calamus rhizomes is evaluated in vitro. Different
concentrations of petroleum ether extract (50 to 2000 mg) are tested and the antimicrobial activity
is observed from 500 mg and the zone of inhibition increases with concentration. The maximum
activity is observed at 2000 mg and the highest concentration tested, beyond which the inhibition
zone does not increase. Among the four types of bacteria tested, high inhibition zone is observed
1037 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

on P. aeruginosa (1.62 cm) followed by S. aureus (1.62 cm). E. coli and B. subtilis show smaller
zone of inhibition (1.34 and 1.04 cm, respectively). MIC test shows that the minimum inhibition
concentration is 0.25 mg/mL for P. aeruginosa, S. aureus, B. subtilis, and 0.5 mg/mL for E. coli
24
. Dry and powdered rhizomes of A. calamus L. were extracted with ultrasonic bath using
dichloromethane as solvent. Various concentrations (0.01 to 0.15%) of the extract were
determined for antifungal activity on PDA agar against Alternaria spp. isolated from leaf spot and
Fusarium spp. isolated from wilt diseases of cruciferous vegetable, as well as Botrytis spp.
isolated from gray mold rot of roses and Septoria spp. isolated from leaf spot of chrysanthemum.
The results indicated that all of the molds examined were sensitive to A. calamus extract.
The growth of all tested fungi was completely inhibited at the concentration of 0.10% upward.
Separation by preparative-TLC and guidance by TLC-bioassay using Cladosporium
cladosporioides as a diagnostic fungus revealed an active compound that was identified as beta -
asarone (cis-1,2,4-trimethoxy-5-(1-propenyl)-benzene) by GC-MS. Modhumita 25 fractionated A.
calamus leaves by cation exchange chromatography and gel filtration and the fraction inhibiting
the hyphal extension of phytopathogens was characterized purified protein was identified as a
class III haem peroxidase with a molecular weight of approx. 32 kDa.
The temperature stability of the enzyme was observed from 5 to 60°C with a temperature
optimum of 36°C. Maximum enzyme activity was registered at pH 5·5. The pH and temperature
optima were corroborated with the antifungal activity of the enzyme. The enzyme was localized
in the leaf epidermal cells and lumen tissues of xylem, characteristic of class III peroxidases. The
toxic nature of the enzyme which inhibited hyphal growth was demonstrated against
phytopathogens, such as Macrophomina phaseolina, Fusarium moniliforme and Trichosporium
vesiculosum. Microscopic observations revealed that distortion in the hyphal structure with
stunted growth, increased volume and extensive hyphal branching.
Asha et al 26 evaluated Antimicrobial activity of A. calamus rhizome and leaf extracts obtained
with different solvents viz., petroleum ether, chloroform, hexane and ethyl acetate against fungal
pathogens and reported that Rhizomes and leaf ethyl acetate extracts exhibited pronounced
antifungal activity with diameter zone of inhibition ranged from 20 to 28 and 18 to 25 mm as well
as anti-yeast activity with diameter zone of inhibition ranged from 22 to 25 and 20 to 23 mm,
respectively. The minimum inhibitory concentration (MIC) of the rhizome and leaf extracts for
antifungal activity measured was 2 to 4 mg/ml, except Penicillium chrysogenum whereas against
yeasts was relatively higher, 4 to 5 and 6 to 8 mg/ml. MIC value for antibacterial activity was
comparatively very high ~16 to 42 mg/ml. In addition, authentic ∞- and β - asarones were also
tested for their antimicrobial potential. Both ∞- and β -asarones exhibited very strong
antimicrobial activities against the fungi and yeasts than those of rhizome and leaf extracts.
The study clearly suggested that A. calamus rhizomes and leaves must possess active principle ∞
and β -asarones which is believed to be responsible for their antimicrobial activities. The
alcoholic extract and essential oil (0.2%) exhibited high in vitro activity against some pathogenic
and nonpathogenic fungi, gram -ve and gram +ve bacterias specifically effective against S.aureus,
E.coli and A.niger but less effect on P.selinium. The essential oil (1:1000v/v) was found to be
effective against citrus decay pathogens like P.digitatum, P.italicum, D.natalensis and A.tenuis
and checked their growth. A.calamus as a constituent in AV/EPP/4(Herbal formulation) was

1038 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

found to be effective in controlling infestation of nymphus and infections by Chipicephalus

sanguineus. Essential oils of a mixture including A. calamus oil had significant anti-bacterial anti-
fungal activities 25-32.
Antihepatotoxic Activity: Studies on rat liver fed with drug acetaminophen showed an increase
in size and enlargement of liver. These changes were reversed by silymarin and also ethanolic
extract of Acorus calamus. Acetaminophen fed animal shoed severe centrilobular necrosis and
fatty infiltration27. Treatment with different doses of ethanol extract of Acorus calamus and
silymarin produced mild degenerative changes and absence of centrilobular necrosis. It was also
observed that serum level of glutamate oxalate transaminase (SGOT), glutamate pyruvate
transaminase (SGPT), alkaline phosphatase (ALP) and total bilirubin were significantly elevated
and protein level were found to be considerably decreased in acetaminophen fed animals.
Antioxidant activity: Palani et al 27 reported that ethanolic extract of Acorus calamus has
antioxidant activity comparable to silymarin. They reported a significant increase in the level of
MDA in the acetaminophen treated rats. Treatment with ethanolic extract of Acorus calamus in
the concentration 250 mg/kg and 500 mg/kg, significantly prevented increase in MDA level.
Acetaminophen treatment caused a decrease in the level of SOD, catalase, GOX, GST, in liver
tissues. The treatment of ethanolic extract of Acorus at the doses of 250 mg/kg and 500 mg/kg
resulted in the significant increase in the level of SOD, catalase, GOX, GST, which was found
comparable to the drug silymarin.
Antioxidant activity: Exposure of rats to acrylamide caused hind limb paralysis in 58% of the
animals on day 10 and decreased behavioural parameters, namely distance travelled, ambulatory
time, stereotypic time and basal stereotypic movements compared with the control group. These
rats also had a decrease in the reduced glutathione (GSH) content and glutathione-S-transferase
(GST) activity in the corpus striatum and an increase in striatal dopamine receptors, as evident by
an increase in the binding of 3Hspiperone to striatal membranes. Treatment with the ethanol:
water (1:1) extract of the rhizomes of A. calamus increased the glutathione content and
glutathione-Stransferase activity in the corpus striatum while insignificant changes were observed
in other parameters. Rats treated with acrylamide and A. calamus extract in combination had a
lower incidence of paralysis (18%) compared with those treated with ACR alone on day 10 of the
experiment. The rats also showed a partial recovery in other behavioral parameters.
The levels of GSH content and GST activity increased in the corpus striatum, while the dopamine
receptors decreased compared with the ACR treated rats. The results suggest that the
neurobehavioral changes produced by ACR may be prevented in the following treatment with A.
calamus rhizomes 28. Epieudesmin has been shown to have antineoplastic activity against the
murine P388 lymphocytic leukemia cell line and several human cancer cell lines (BXPC-3, MCF-
7, SF268, NCI-H460, KM20L2, and DU- 145). Galgravin has demonstrated activity in preventing
neuronal death and stimulating neurite growth. Structurally, similar lignans have also shown
neuroprotective activity in in vitro models for Alzheimer’s and Parkinson’s disease. Both
epieudesmin and galgravin were identified in the methanolic extracts of A calamus leaves by
liquid chromatography electron impact mass spectrometry 22.
The steam volatile fraction of the roots and rhizomes of A. catamus prolongs the sleeping time of
mice when used with pentobarbital, hexobarbital and ethanol. It reduces body temperature of

1039 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

mice. The maximum reduction of body temperature and the potentiation of the hypnotic activity
are observed 1 h after its administration. It exacerbates tonic seizures provoked by convulsive
doses of Metrazol in rats and potentiates of the action of reserpine in reducing amphetamine
toxicity in aggregated mice. The fall produced in the blood pressure of anesthetized rats is not
prevented by vagal, adrenengic or ganglionic blockade and does not appear to be due to any
nervous mechanism. It causes dilatation of the blood vessels of the splanchnic area in cats and
constricts the blood vessels of the frog's hind limbs. It prevents the action of acetylchohine,
histamine and barium chloride on the isolated guinea pig's ileum.
Anti-itching: Boiling water extract of A.calamus is used as bathing agent for skin diseases.
A.calamus was used as one of the constituent in a poly herbal formulation namely, herbal multi
action skin gel (AV/AAGD/14/), which was found to be effective against a variety of specific and
nonspecific dermatitis and maggot wounds 29, 30.
Anti-anxiety: Three drugs i.e. Brahmi, Vacha and Shankhapuspi used in combination as ratio
10:3.8:0.2 exhibits a significant antianxiety activity 35. Both the poly herbal formulations having
A.calamus as an ingredient namely, Prasham (100mg) and P-tabs significantly provides a good
relief against insominia, stress excitement and irritability 31, 32.
Anti-convulsant: A.calamus oil (100mg/kg) was ineffective against metrazol and minimal
electro shock induced seizures in albino mice 33. Plant extract of Acorus calamus did not afford
strychnine induced convulsion at 10-20mg/kg. It caused –ve ionotropic and chronotropic effects
in frogs at 100µ/ml. It antagonized spontaneous motor activity and also amphetamine induced
hyperactivity in mice 34.
Anti-viral and Anti-anginal activity: The alcoholic extract of A.calamus exhibited potent
antiviral activity against herpes virus.i.e.HSV-1 and HSV- 2 40. A. Calamus in dose of 1.5
3gm/day was found effective against ischaemic heart disease, improvement in chest pain, instable
angina, dyspnoae reduction of body weight, improving in ECG, decreasing serum cholesterol,
decreasing SLDL and increasing SHDL41. A. calamus being a constituent in polyherbal drug
namely Haritaki vati (HT), which reduced the anginal frequency and decreases the serum
cholesterol and serum triglyceride levels 35.
Anti-ulcer: The ethanolic extract of acorus rhizome is used as antiulcer agent which inhibits the
gastric secretion and to protect gastrodudonal mucosa against the injuries caused by pyloric
ligation in rat 36.
Anti-spasmodic: At a dose level of 10µg/ml, the _-asarone free oil (type-I) of A.Calamus had a
pronounced spasmolytic activity 37, 38.
Anti-inflammatory: The aqueous extract of A.calamus showed a significant caraigeenan induced
anti-inflammatory activity at a low dose 39-41.
Anti-cancer Activity: α-asarone isolated from the volatile fraction of A. Calamus was found to
exhibit anti-carcinogenic action on ED50 of SGC Cells at a dose level of 25mcg/ml 42.
Nootropic activity: α-asarone isolated from the volatile fraction of A. Calamus was found to
exhibit anti-carcinogenic action on ED50 of SGC Cells at a dose level of 25mcg/ml 43.

1040 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

Anti-sczhizophrenia: GK022, an herbal mixture containing A.calamus (100mg) as a constituent

was reported to be significant in cases of Schizophrenia 44.
Tranquilizer: The aqueous extract was also supposed to counteract the effect of mental stress by
tranquilizing action as mentioned in ayurvedic texts 45.
Anti-asthmatic: Small pieces of rhizome (1kg) and an ayurvedic receipe namely Maduyashtyadi
syrup in which A. Calamus as an ingredient, after administration showed a significant relief in
bronchospasm without any side effect 46.
Anti-rheumatitis: Vachadi gana, which consists of six plants including A. Calamus was found to
be effective in case of Rheumatoid arthritis with pain, swelling and functional disability.

CULTURAL ASPECTS OF ACORUS CALAMUS

Soil and climate: It is a hardy plant found growing from tropical to subtropical climates. Plenty
of sunshine should be available to the plant during its growth and after harvesting for drying the
rhizomes. Temperature ranging from 10 to 38°C and annual rainfall between 70 and 250 cm are
best suited. Cultivation should be avoided in places where there is no irrigation facility. This
species comes up well in clayey loams, sandy loams and light alluvial soils of river banks.

LAND PREPARATION

The land should be ploughed two to three times just before the onset of rains. The land should be
prepared like paddy fields.
Propagation: Acorus is propagated through rhizomes. Rhizomes obtained from earlier planting
are kept preserved in the soil and constantly kept moist. After emergence, the rhizomes are cut
into small pieces and planted. Sprouted rhizome pieces are planted at a spacing of 30 x 30 cm and
depth of 4 cm in the month of July-August. The best time for planting is the second fortnight of
June. Around 1, 11,000 plants can be planted per hectare. As the growth rate is very fast, sprouts
are visible on the second day of planting.
Fertilizers: Compost/FYM @15 t per hectare along with nitrogen and phosphorus is applied.
One third of N along with 50 kg of P and 25 kg of K is the basal requirement. The second dose of
N should be given after one month of planting as broadcast and a third dose should be applied
after two months of planting.
Irrigation: The river or canal banks where the land is saturated with water is very suitable for its
growth. The initial level of water standing in the field should be 5 cm and later increased to 10
cm. Irrigation can be avoided in the rainy season, however, if there is prolonged dry spell it must
be irrigated at an interval of 2 to 3 days.
Plant protection: Mealy bugs and caterpillar are the pests occurring on this crop. Spraying the
shoots and drenching the roots of plants with 10 ml methyl parathion or 20 ml Quinolphos in 10
L of water can be effective in controlling the shoot and root mealy bugs. Major disease is leaf
spot and a spray of Captan 10 g with Chloropyriphos 20 ml/10 L controls leaf spot as well as
mealy bugs and caterpillar.

1041 J. Chem. Bio. Phy. Sci. Sec. B, May 2016 – July 2016; Vol.6, No.3; 1027-1045
Acorus Calamus L … Ranjan et al.

Harvesting and post-harvest operations: After 6 to 8 months, in December, the lower leaves
turn yellow and dry indicating their maturity. The field should be partially dried only leaving
sufficient moisture for uprooting the plant. In case of large scale cultivation rhizomes may be
removed by passing the plough. The uprooted rhizome is cleaned after washing with water and
cut into size of 5 to 7.5 cm length and fibrous roots removed. The cut rhizomes are dried by
spreading under the shade so that the amount of oil present in it is not harmed.
Yield: The yield is expected to be 4.22 t of dry rhizomes or 10 t fresh rhizomes per hectare.
Insecticidal activity: Asarones (2, 4, 5-trimethoxypropenyl-benzenes) isolated from the essential
oil of A. calamus L. rhizomes, are potent growth inhibitors and anti-feedants to the variegated
cutworm. cis-Asarone added to artificial diet significantly inhibited growth and feeding by first-,
third-, and fourth-instar larvae, whereas the trans isomer produced an anti-feedant effect alone.
Gross dietary utilization (efficiency of conversion of ingested food, ECI) was decreased when the
diet was supplemented with cisasarone or when this compound was topically applied to fourth-
instar larvae.
Inhibition of growth occurred even as a moderate topical dose (5 mu g/larva) primarily as a result
of decreased efficiency of conversion of digested food (ECD), even though the approximate
digestibility (AD) of the food was unchanged. The insecticidal activities of compounds derived
from the rhizomes of A. gramineus against four agricultural insect pests were examined using
direct contact application method. The biologically active constituents of A. gramineus rhizomes
were characterized as the phenylpropenes, cis- and trans-asarones by spectroscopic analyses.
Potencies varied according to insect species, compound, and dose.
In a test with female adults of Nilaparvata lugens, cis-asarone caused 100, 83 and 40% mortality
at 1,000, 500 and 250 ppm, respectively, whereas 67% mortality was achieved at 1,000 ppm of
trans-asarone. Against 3rd instar larvae of Plutella xylostella, cis-asarone gave 83 and 50%
mortality at 1,000 and 500 ppm, respectively, whereas transasarone at 1,000 ppm showed 30%
mortality. Against female adults of Myzus persicae and 3rd instar larvae of Spodoptera litura, cis-
and trans-asarones both were almost ineffective at 2,000 ppm. The A. gramineus rhizome-derived
materials merit further study as potential insect-control agents or as lead compounds against N.
lugens and P. xylostella.

CONCLUSION

Even though, this studies supports the different pharmacological activities of Sweet flag lot of
clinical experiments will have to be conducted in future to exploit the full potential activities of
this crop and this plant species has to properly identified and conserved to avoid the extinct
condition.

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*Corresponding author: Alok Ranjan;

Department of Botany, GPGC Obra, Sonbhadra, U.P. (India)

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