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Advances in Biological Research 1 (1-2): 17-21, 2007
ISSN 1992-0067
© IDOSI Publications, 2007
Corresponding Author: Dr. B.D. Ranjitha Kumari, Stress Physiology and Medicinal Plant Biotechnology Unit, Department of
Plant Science, School of Life Sciences, Bharathidasan University,
Tiruchirappalli - 620 024, Tamil Nadu, India
17
Phytochemical and Antibacterial Studies of Chicory
(Cichorium intybus L.) - A Multipurpose Medicinal Plant
S. Nandagopal and B.D. Ranjitha Kumari
Stress Physiology and Medicinal Plant Biotechnology Unit, Department of Plant Science,
School of Life Sciences, Bharathidasan University, Tiruchirappalli-620 024, Tamil Nadu, India
Abstract: Chicory (Cichorium intybus L.) belongs to the family Asteraceae and it is a small aromatic biennial
or perennial herb. The whole plant contains a number of medicinally important compounds such as inulin,
esculin, volatile compounds (monoterpenes and sesquiterpenes), coumarins, flavonoids and vitamins. In the
present study, we evaluated the phytochemical analysis for the presence of various secondary metabolites and
antibacterial activity of the root extracts of chicory against pathogenic bacteria like gram positive (Bacillus
subtilis, Staphylococcus aureus and Micrococcus luteus) and gram negative (Escherichia coli and Salmonella
typhi) bacteria by in vitro agar well diffusion method. The hexane and ethyl acetate root extracts of chicory
showed pronounced inhibition than chloroform, petroleum ether and water extracts. Root extracts showed more
inhibitory action on Bacillus subtilis, Staphylococcus aureus and Salmonella typhi than Micrococcus luteus
and Escherichia coli.
Key words: Cichorium intybus L.
antibacterial activity root extracts zone of inhibition volatile
compounds
INTRODUCTION arthralgia, lumbago, asthma and general debility [4, 5].
Higher and aromatics plants have been used
traditionally in folk medicine as well as to extend the shelf
life of foods, showing inhibition against bacteria, fungi
and yeasts [1]. Biologically active compounds from
natural sources have always been a great interest for
scientists working on infectious diseases [2]. Cichorium
intybus L. is a medicinally important plant that belongs to
the family Asteraceae. The tuberous root of this plant
contains number of medicinally important compounds
such as inulin, bitter sesquiterpene lactones, coumarins,
flavonoids and vitamins [3]. The plant root is used as
antihepatotoxic, antiulcerogenic, antiinflammatory,
appetizer, digestive, stomachic, liver tonic, cholagogue,
cardiotonic, depurative, diuretic, emmenagogue,
febrifuge, alexeteric and also as tonic. It is useful in
vitiated conditions of kapha and pitta, cephalalgia,
heapatomegaly, inflammations, anorexia, dyspepsia,
flatulence, colic, gout, burning sensation, allergic
conditions of skin, jaundice, splenomegaly, hyperdipsia,
skin diseases, leprosy, strangury, amenorrhoea, chronic
and bilious fevers, ophthalmia, pharyngitis, vomiting,
This plant is also used to treat AIDS, cancer, diabetes,
dysmenorrhoea, impotence, insomnia, splenitis and
tachicardia [6]. Inulin is used to replace fat or sugar and
reduce the calories of food. It is suitable for consumption
by diabetics [7] and is also used in inulin clearance test
to measure glomerular filtration rate-GFR [8]. Recent
pharmacological investigation of the root extract of
this plant revealed immunomodulator, antitumor and
anticancer properties [9, 10]. The root is rich in alkaloids,
which forms an ingredient or adulterant in coffee. The
deep purple flower heads yield blue dye. The flowers
are also used in floral clocks by Linnaeus [11]. The
sesquiterpene lactones such as lactucin and lactucopicrin
were isolated from chicory and reported for its
antibacterial and antimalarial activity [12]. The antifungal
activity of chicory was also reported [13-16]. Based on the
studies carried out in chicory, world wide report shows
that the roots and leaves of this plant possess strong
antibacterial and nematicidal effect [17]. However, to the
best of our knowledge, very few reports are available on
antibacterial properties of chicory root against the
important human pathogenic bacteria so far. In the
Advan. Biol. Res., 1 (1-2): 17-21, 2007
18
present study we reported the antibacterial activity of Determination of antibacterial activity: The antibacterial
Cichorium intybus L. against pathogenic bacteria. The activity of the root extracts was determined using
study confirms that both aqueous as well as organic agar well diffusion method by following the published
solvent root extracts possess strong antibacterial procedure with slight modification [22]. Nutrient agar was
properties against various pathogens such as gram inoculated with the given microorganisms by spreading
positive (Bacillus subtilis, Staphylococcus aureus and the bacterial inoculums on the media. Wells (8 mm
Micrococcus luteus) and gram negative (Escherichia diameter) were punched in the agar and filled with plant
coli and Salmonella typhi) bacteria. extracts. Control wells containing neat solvents (negative
MATERIALS AND METHODS control) viz., chloromphenicol (50 and 100 µg ml
)
Preparation of root extracts: Apparently healthy plant were incubated at 37°C for 18 h and the antibacterial
roots were collected, washed thoroughly in tap water and activity was assessed by measuring the diameter of
dried in room temperature for 15 days. The dried 25 g root the zone of inhibition. The relative antibacterial potency
was powdered and soaked separately in 100 ml petroleum of the given preparation was calculated by comparing
ether, hexane, chloroform, ethyl acetate and water by its zone of inhibition with that of the standard drug
keeping it in a shaker for 3 days. The extracts were filtered chloromphenicol.
through cheesecloth and the extracts were reduced to
10% of its original volume. The organic solvent filtrates Statistical analysis: The resultant clear zones around
were concentrated in vacuum using a rotary evaporator, the discs were measured in mm. The antibacterial
while aqueous extract was dried using water bath. activity of chicory root extracts was indicated by clear
Phytochemical screening of root extracts: The experiments represented by five replicates from each
phytochemical components of the chicory roots were experiment were subjected to statistical analysis
screened by using the methods of Brindha et al. [18] (Mean±SE), according to New Duncan’s Multiple Range
and Harbone [19]. The components analysed were Test [24].
alkaloids, volatile oils, fatty acids, emodins, flavonoids,
triterpenoids, antharacene glycosides, tannins, phenolics RESULTS
and saponins.
Separation of the compounds: The compounds present preliminary phytochemical screening of the root extracts
in chicory root extracts were qualitatively analyzed by using different solvents was reported (Table 1). All the
using thin layer chromatography which is commercially
available; TLC aluminum sheets with silica gel 60F were
254
used. The isolation and separation of monoterpenes
and sesquiterpenes was done by using the procedure
of Brindha et al. [18] and Janusz Malarz [20].
Inoculums: The test microorganisms, gram positive
(Bacillus subtilis, Staphylococcus aureus and
Micrococcus luteus) and gram negative (Escherichia
coli and Salmonella typhi) bacteria were obtained from
culture repository of Best Biotech culture collection,
Bangalore, India. The organisms were inoculated into
NB (Nutrient Broth) medium, (0.5% Peptone, 0.5%
Sodium Chloride, 0.15% Yeast extract; pH 7.4) and
incubated at 37°C for overnight. The bacterial cells were
harvested by centrifuging at 5000g for 15 min. The pellet
formed was washed twice with PBS (Phosphate Buffer
Saline), (10 mM Sodium Chloride, pH 7.4) and the cells
were counted by hemocytometer. The bacterial cells were
diluted to approximately 10 CFU ml
before use [21].
51
control) or standard antibiotic solution (positive
1
were also run parallel in the same plate [23]. The plates
zones of growth inhibition. Data of three independent
Phytochemical screening of root extracts: The
Table 1: Preliminary phytochemical analysis of chicory root extracted with
different solvents
Name of the
S. No. compound PE C H EA W
1 Alkaloids - - - + +
2 Volatile oils + + + + -
3 Fatty acids + + + + -
4 Emodins - - - - -
5 Flavonoids - - - - +
6 Triterpenoids - - + - -
7 Antharacene glycosides - - - - -
8 Tannins - - - - +
9 Phenolics - - - - -
10 Saponins + + - + +
+ Present, -Absent, PE-Petroleum ether, C-Chloroform, H-Hexane, EA-
Ethyl acetate, W-Water
Advan. Biol. Res., 1 (1-2): 17-21, 2007
19
Table 2: Antibacterial activity of root extracts of chicory at 50 µl concentration
Zone of inhibition in mm (Mean±SD)
---------------------------------------------------------------------------------------------------------------------------------------------------------
Gram(+) Gram(-)
------------------------------------------------------------------------------------- -----------------------------------------------
Solvent B. subtilis S. aureus M. luteus E. coli S. typhi
Petroleum ether 6.8±0.17 5.8±0.21 4.2±0.13 4.7±0.41 3.5±0.31
cd c d d e
Chloroform 4.1±0.25 3.6±0.23 2.6±0.28 3.0±0.28 5.4±0.32
ef d e e c
Hexane 12.3±0.21 9.8±0.03 8.0±0.41 9.6±0.32 7.5±0.44
a a a aa
Ethyl acetate 10.5±0.28 2.0±0.27 6.8±0.30 6.3±0.41 3.0±0.25
b e b bc ef
Water 4.8±0.25 2.8±0.23 3.3±0.23 1.9±0.35 4.5±0.33
e de de ef d
chloromphenicol 8.4±0.23 9.6±0.23 6.0±0.14 7.5±0.29 7.3±0.30
c ab bc b ab
The negative control wells were exposed with the neat solvent and the positive control was chloromphenicol (50 µg ml ). Each value represents the
1
mean±standard error (SE) of five replicates per treatment in three repeated experiments. Means within a row followed by the same letters are not significant
at p = 0.05 according to DMRT
Table 3: Antibacterial activity of root extracts of chicory at 100 µl Concentration
Zone of inhibition in mm (Mean±SD)
---------------------------------------------------------------------------------------------------------------------------------------------------------
Gram(+) Gram(-)
------------------------------------------------------------------------------------- -----------------------------------------------
Solvent B. subtilis S. aureus M. luteus E. coli S. typhi
Petroleum ether 14.3±0.39 10.10±0.16 5.8±0.33 8.0±0.33 5.7±0.32
c c d cd de
Chloroform 10.5±0.21 8.20±0.22 4.8±0.32 3.2±0.30 14.0±0.31
d cd de f b
Hexane 18.2±0.47 18.23±0.18 15.2±0.04 14.5±0.22 19.2±0.43
a a a aa
Ethyl acetate 15.0±0.17 3.60±0.20 13.6±0.29 10.1±0.26 10.7±0.30
bc e ab c c
Water 8.0±0.27 6.70±0.28 10.3±0.56 5.6±0.23 5.8±0.28
e d c ed
chloromphenicol 16.0±0.18 17.30±0.37 13.0±0.33 13.5±0.37 14.0±0.23
b ab b ab b
The negative control wells were exposed with the neat solvent and the positive control was chloromphenicol (100 µg ml ). Each value represents the
1
mean±standard error (SE) of five replicates per treatment in three repeated experiments. Means within a row followed by the same letters are not significant
at p = 0.05 according to DMRT
four organic solvents such as petroleum ether, chloroform Micrococcus luteus) and gram negative (Escherichia coli
hexane and ethyl acetate showed positive result for the and Salmonella typhi) bacteria. At 50 µl concentration,
presence of volatile oils and fatty acids which were absent the hexane extract showed pronounced inhibition against
in the water extract. The ethyl acetate and water extracts all the tested organisms, the maximum inhibition was
showed the presence of alkaloids. Only in the hexane observed on B. subtilis (12.3±0.21 mm), S. aureus
extract presence of triterpenes was observed. In the water (9.8±0.03 mm) and E. coli (9.6±0.32 mm) and moderate
extract, flavonoids, tannins and saponins were present inhibition was observed on M. luteus (8.0±0.41 mm) and
which were absent in the organic solvent extracts. S. typhi (7.5±0.44 mm). The other solvents extracts did
Antibacterial activity of different solvent extracts: We concentration except ethyl acetate against B. subtilis
used both polar as well as non-polar solvents for the (10.5±0.21 mm) (Table 2). The growth of B. subtilis was
extraction of active components from the roots of chicory inhibited by all the root extracts at 100 µl concentration
plant. The antibacterial activity of the chicory roots was and maximum inhibition was observed with hexane
assessed using the agar well diffusion method by extract as 18.2±0.47 mm zone of inhibition, which was
measuring the diameter of growth inhibition zones with higher than the zone of inhibition caused by the standard
50 and 100 µl of different solvent extracts (Table 2 and 3). drug chloromphenicol (16.0±0.18 mm). Similarly, hexane
The results showed that all the five solvent extracts root extract produced maximum inhibition (18.0±0.18 mm)
possesses antibacterial activity against the tested gram to the growth of S. aureus than chloromphenicol
positive (Bacillus subtilis, Staphylococcus aureus and (17.3±0.37 mm) whereas other extracts showed less
not actively inhibit the growth of the bacteria at 50 µl
Advan. Biol. Res., 1 (1-2): 17-21, 2007
20
inhibitory activity than chloromphenicol. The hexane Staphylococcus aureus and Micrococcus luteus) and
(15.2±0.04 mm) and ethyl acetate (13.6±0.29 mm) extracts gram negative (Escherichia coli and Salmonella typhi)
were effective against M. luteus and the water extract bacteria. The present report indicates that increased
(10.3±0.56 mm) showed less inhibition than the standard lipophillic compounds are extracted using the petroleum
drug whereas the other two extracts showed very ether, chloroform and hexane increased the suspended
minimum inhibition on the growth of M. luteus. The higher compounds in the above solvents as stated [26].
hexane (14.5±0.22 mm) and ethyl acetate (10.1±0.26 mm) Chicory plant was active against gram positive and gram
root extracts showed maximum inhibition and petroleum negative bacteria, yeast and filamentous fungi [17]. The
ether extract (8.0±0.33 mm) showed moderate inhibition antibacterial activity of water, ethanol and ethyl acetate
and the water (5.6±0.23 mm) and chloroform (3.2±0.30 mm) extracts of chicory was reported against Agrobacterium
extracts showed less inhibition on E. coli. S. typhi was radiobacter, Erwinia carotovora, Pseudomonas
inhibited by all the root extracts and the maximum fluorescens and Pseudomonas aeruginosa [27]. The
inhibition was observed with hexane (19.2±0.43 mm), present work shows that the compounds from chicory
chloroform (14.0±0.31 mm) and ethyl acetate (10.7±0.30 possess potent antimicrobial activity and suggesting that
mm) whereas petroleum ether (5.7±0.32 mm) and water the chicory root extracts contains the effective active
(5.8±0.28 mm) extracts showed least inhibition at 100 µl constituents responsible for eliminating the bacterial
concentration (Table 3). We found that both aqueous pathogens. Finally, it can be concluded that the active
as well as organic extracts of the roots were successful chemical compounds present in chicory (Cichorium
in inhibiting the bacteria in a dose dependent manner. intybus) should certainly find place in treatment of
Besides the 50 µl concentration of root extracts, the various bacterial infections. The results of this study are
100 µl concentration of root extracts were found to very encouraging and indicate that this herb should be
possess maximum inhibition (Table 2 and 3). studied more extensively to explore its potential in the
DISCUSSION
We used both the aqueous and organic solvents
for the extraction of active components from roots of 1. Hulin, V., A.G. Mathot, P. Mafart and L. Dufosse,
the chicory plant. The result of the study reveals that 1998. Les proprietes anti-microbiennes des huiles
the non-polar and polar solvent extracts were active essentielles et composes daromes. Sci. Aliments, 18:
against the strains of the bacteria that are common cause 563-582.
of infections. Chicory shows significant antibacterial 2. Perumal Samy R. and S. Ignacimuthu, 2000.
activity that may be due to the presence of many Antibacterial activity of some medicianal plants
potent compounds such as inulin, bitter sesquiterpene from Eastern Ghats, South India. Solai Bull.
lactones, coumarins, flavonoids etc. The biologically Ethnopharmacol., 72: 39-41.
active compounds are screened by dissolving the crude 3. Varotto, S., M. Lucchin and P. Parrin, 2000. Immature
powder on various compound specific solvents confirmed embryos culture in Italian red Chicory (Cichorium
by the TLC (data not shown). The antibacterial activity intybusc). Plant Cell Tiss. Org. Cult., 62: 75-77.
was expressed at varying degrees which was being both 4. Nadkarni, A.K., 1976. Indian Materia Medica popular
strain and dose dependent. The various crude extracts of Prakasam Pvt. Ltd., Bombay.
chicory showed significant activity against all the bacteria 5. Rastogi, R.P. and B.N. Mehrotra, 1994. Compendium
tested. Similar to our results, the biological activity of of Indian Medicinal Plants. Rastogi, R.P. (Eds.).
Mentha piperita against the pathogenic bacteria was Orient Longman Ltd., Madras, pp: 74.
reported [25]. We used five different solvent extracts of 6. Duke, J.A., 1983. Medicinal Plants of the Bible.
chicory roots which showed activity against all bacterial (Illustrated by Peggy K. Duke) Out of print Trado-
species tested at all the dosages. The root extracts of Medic Books Buffalo and NY., pp: 233.
chicory exhibited antibacterial activity on hexane, ethyl 7. Niness, K.R., 1999. Inulin and Oligofructose: What
acetate, petroleum ether, chloroform and aqueous extracts Are They? J. Nutr., 129: 1402S-1406S.
against the bacteria tested at 50 µl-100 µl concentrations 8. Vasudevan, D.M. and S. Sreekumari, 1995. Text
by following the method of Valsaraj et al. [23]. We Book of Biochemistry for MedicalStudents. 2nd Ed.
observed maximum activity at 100 µl concentration Jaypee Brothers Medical Publisher (P) Ltd., New
against the tested gram positive (Bacillus subtilis, Delhi, India.
treatment of many infectious diseases.
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