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INDEX – GJRMI, Vol.2, Iss. 2, February 2013
Medicinal plants Research
Natural Resource
CHEMICAL COMPOSITION AND ANTIBACTERIAL ACTIVITY OF ESSENTIAL OIL OF
ZIZIPHORA HISPANICA L.
Bounar Rabah, Takia Lograda, Messaoud Ramdani, Pierre Chalard and Gilles Feguiredo 73–80
Agriculture
COMPARISON OF COLCHICINE CONTENT BETWEEN HYSTERANTHOUS AND
SYNANTHOUS COLCHICUM SPECIES IN DIFFERENT SEASONS
Alirezaie Noghondar Morteza, Arouee Hossein, Shoor Mahmoud, and Rezazadeh Shamsali 81–88
Biological Science
ECOLOGICAL AND MEDICINAL INTEREST OF TAZA NATIONAL PARK FLORA (JIJEL -
ALGERIA)
BOUNAR Rabah, REBBAS Khellaf, GHARZOULI Rachid, DJELLOULI Yamna and ABBAD abdelaziz
89–101
Bio-Technology
CONSERVATIVE PRODUCTION OF BIODIESEL FROM WASTE VEGETABLE OIL Chethana G S, Reddy K Dayakar, Vijayalakshmi 102–109
Indigenous medicine
Ayurveda
PHYTOCHEMICAL STUDIES ON SMILAX MACROPHYLLA LINN.; A SOURCE PLANT OF
CHOPACHEENI
Jyothi T, Acharya Rabinanaryan, Shukla C P, Harisha CR 110–117
SELECTION OF MEDICINAL PLANTS FOR THE MANAGEMENT OF DIABETIC FOOT
ULCER; AN AYURVEDIC APPROACH
Pampattiwar S P, Adwani N V, Sitaram Bulusu, Paramkusa Rao M 118–125
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – FRUITS OF MALLOTUS PHILIPPENSIS (LAM.) MULL. ARG,
OF THE FAMILY EUPHORBIACEAE PLACE – AGUMBE, SHIMOGA DISTRICT, KARNATAKA, INDIA
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
CHEMICAL COMPOSITION AND ANTIBACTERIAL ACTIVITY OF
ESSENTIAL OIL OF ZIZIPHORA HISPANICA L.
Bounar Rabah1, 6
, Takia Lograda2*, Messaoud Ramdani
3, Pierre Chalard
4 and
Gilles Feguiredo5
1, 2, 3Laboratory of Natural Resource Valorization, Sciences Faculty, Ferhat Abbas University, 19000 Setif,
Algeria 4Clermont Université, Université Blaise Pascal, BP 10448, F-63000 Clermont Ferrand
5LEXVA Analytique, 460 rue du Montant, 63110 Beaumont, France
6Department of Natural Sciences and Life, Faculty of Science, M'sila University, 28000 M’sila (Algeria)
*Corresponding Author : [email protected]; +21336835894, +213 776243824; Fax : +21336937943
Received: 24/12/2012; Revised: 25/01/2013; Accepted: 31/01/2013
ABSTRACT
The aerial parts of Ziziphora hispanica L. species were collected on April 2011 from Boussaâda
localities in Algeria. The chemical compounds of the plant were isolated by hydrodistillation. A total
of 28 constituents, representing more than 93.8% of the total oil, were identified by gas chromato-
graph/mass spectrometry (GC/MS). The most presented compounds of the essential oil of Z. his-
panica were Pulegone (78.6%), limonene, menthofuran, trans-iso-pulegone and piperitenone are rep-
resented by low concentrations. The essential oil of aerial parts of Z. hispanica has a broad spectrum
of antimicrobial activity. The sensitivity of bacteria and fungi tested with essential oil compounds
was found to be very high.
Key words: Ziziphora hispanica L., essential oil, Antibacterial activity, Algeria
Research article
Cite this article:
Bounar R, Takia L, Messaoud R, Pierre C and Gilles F (2013), CHEMICAL COMPOSITION AND
ANTIBACTERIAL ACTIVITY OF ESSENTIAL OIL OF ZIZIPHORA HISPANICA L.,
Global J Res. Med. Plants & Indigen. Med., Volume 2(2): 73–80
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Belonging to the family Lamiaceae,
Ziziphora hispanica L. is an annual plant with
very branchy erect stem. The leaves are all
similar, ovate-lanceolate, and ciliate on
margins. Spike-like inflorescences composed of
verticillastres pauciflores; corolla long tubular
structure (Quézel et Santa, 1962–1963). This
plant is found in areas of the Saharan Atlas and
the highlands. Some of Ziziphora species are
used for their aperitif, carminative and
antiseptic effects in treatment of various
diseases (Ozturk and Ercisli, 2007), especially
Z. taurica infusions (Tzakou et al., 2001;
Gözde et al., 2006). Z. persica is an edible
medicinal plant, it is frequently used as wild
vegetable or additive in foods to offer aroma
and flavour (Nezhadali et al., 2008, 2009,
2010). Z. clinopodioides, riche in monoterpene
glucosides (Megumi et al., 2012), is used
mostly in food and medicine (Maya, 2012). Z.
hispanica is used as a substitute for Morocco
pennyroyal (Mentha pulegium) (Bellakhdar,
1997). According to the population of
Boussaâda Z. hispanica is used as an infusion
to soothe the stomach pains, for the heart
fatigue and added to the coffee for a better
taste.
A literature survey showed that the oil of
Ziziphora species has been found to be rich in
pulegone. The major components in Z. taurica,
Z. vychodceviana and Z. persica are pulegone
and isomenthone (Dembistikii et al., 1995;
Sezik and Tumen, 1990; Nezhadali and
Zarrabi, 2010). The major constituent found in
the oil of Z. tenuior L. has been reported to be
pulegone (Sezik et al., 1991). The chemical
composition of Z. clinopodioides Lam. was
analyzed, the major constituents were pulegone
and piperitenone (Salehi et al., 2005; Sonbola
et al., 2006; Verdian-Rizi, 2008; Xing et al.,
2010; Soltani, 2012). The essential oil of
Turkish Z. taurica subsp. clenioides was found
to contain pulegone (Meral et al., 2002). The
major constituents of essential of Z.
pamiroalaica were pulegone and menthone
(Xing et al., 2010). Z. capitata contained no
oil; Z. persica, Z. taurica, Z. Tenuior and Z.
clinopodioide have a Pulegone as a major
compound while Z persica has a major
component the thymol (Hüsnü, 2002). The oil
of Z. hispanica is characterised by pulegone
(Velasco and Mata, 1986; Bellakhdar, 1997;
Bekhechi et al., 2007).
The essential oil of Ziziphora species has a
broad spectrum of antimicrobial activity. The
oil of Z. clinopodioides was found to exhibit
interesting antibacterial activity against
Staphylococcus epidermidis, S. aureus,
Escherichia coli and Bacillus subtilis (Sonbola
et al., 2006), the oil of Z. clinopodioides was
tested against some human pathogenic bacteria,
which showed good activity against all tested
bacteria, except for Pseudomonas aeruginosa
(Soltani, 2012). Investigation of the
antimicrobial activity of the essential oil of the
Turkish endemic Ziziphora taurica on eight
bacterial strains and Candida albicans, indicate
that the essential oil remarkably inhibited the
growth of tested microorganisms except
Candida albicans (Gozde et al., 2006). Z.
tenuior oils had bactericidal and inhibitory
effects of K. pneumoniae, It can be used as
candidates for treatment of infectious diseases
that is caused by this bacteria (Mahboubi et al.,
2012). The oil from Z. pamiroalaica was better
than that from Z. clinopodioides in antioxidant
abilities (Xing et al., 2010). The insecticidal
and ovicidal effects of essential oil of Z.
clinopodioides were tested on adults and eggs
of Callosobruchus maculatus (Lolestani and
Shayesteh, 2009). The objective of this
research is to determine the chemical
composition of essential oil of Z. hipanica from
the Boussaada region and evaluate its potential
to be antimicrobial.
MATERIALS AND METHODS
Plant material
Aerial parts of Ziziphora hispanica were
collected during the flowering stage in October
2011 from Boussaâda localities in Algeria.
Identified by Dr. Lograda Takia, the voucher
specimen is deposited in the herbarium of the
Department of Biology, Ferhat Abbas
University, Algeria. Z. hispanica was submitted
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
to hydrodistillation for 3h using a Clevenger
apparatus (Lograda et al., 2013). The distilled
essential oils were stored at +4 °C for further
use.
Essential oil Analysis:
The essential oils were analysed on a
Hewlett-Packard gas chromatograph Model
5890, coupled to a Hewlett-Packard model
5971, equipped with a DB5 MS column (30 m
X 0.25 mm; 0.25 μm), programming from 50°C
(5 min) to 300°C at 5°C/min, with a 5 min
hold. Helium was used as the carrier gas
(1.0 mL/min); injection in split mode (1:30);
injector and detector temperatures, 250 and
280°C, respectively. The mass spectrometer
worked in EI mode at 70 eV; electron
multiplier, 2500 V; ion source temperature,
180°C; MS data were acquired in the scan
mode in the m/z range 33–450. The
identification of the components was based on
comparison of their mass spectra with those of
NIST mass spectral library (Masada, 1976;
NIST, 2002) and those described by (Adams,
2001) as well as on comparison of their
retention indices either with those of authentic
compounds or with literature values (Adams,
2001).
Antibacterial activity:
Two Gram positive bacteria (Staphylococ-
cus aureus ATCC25923 and Bacillus subtilis
ATCC 6633) and seven Gram negative bacteria
(Pseudomonas aeruginosa ATCC27853, Pseu-
domonas syringae pv. Tomato ATCC 1086;
Escherichia coli ATCC 25922, klebsiella
pneumoniae CIP 53-153, Salmonella enterica
CIP 60-62T, Enterobacter sp. and Citrobacter
sp.) and three fungi (Aspergelus flavus
LBVM20, Aspergilus niger LBBM62 and
Candida albicans ATCC 24433) were used in
this study. The bacterial inoculums was pre-
pared from overnight broth culture in physio-
logical saline (0.8 % of NaCl) in order to obtain
an optical density ranging from 0.08–01 at 625
nm. Muller-Hinton agar (MH agar) and MH
agar supplemented with 5% sheep blood for
fastidious bacteria were poured in Petri dishes,
solidified and surface dried before inoculation.
Sterile discs (6 mm Φ) were placed on inocu-
lated agars, by test bacteria, filled with 10 μl of
mother solution and diluted essential oil (1:1,
1:2, 1:5, and 1:10 v:v of Dimethylsulfoxide
(DMSO). DMSO was used as negative control.
Chloramphenicol for bacteria and amphotericin
B for fungi were used as positive control. Bac-
terial growth inhibition was determined as the
diameter of the inhibition zones around the
discs. All tests were performed in triplicate.
Then, Petri dishes were incubated at 37°C dur-
ing 18–24 h aerobically (Bacteria) and at 25°C
for 7 days (fungi). After incubation, inhibition
zone diameters were measured and docu-
mented.
RESULTS
The essential oil, of Ziziphora hispanica L.,
isolated by hydrodistillation from the aerial
parts, was obtained in yield of 1.01% (v/w).
The chemical composition of essential oil, ana-
lyzed by gas chromatography/mass spectrome-
try (GC-MS), gave 28 constituents representing
93.82% of the total oil. The names of the corre-
sponding compounds and their percentages are
listed in table 1.
The oil is characterized by a high content of
pulegone (78.6%). Other compounds a low rate
were piperitenone (2.9%), 8-hydroxy-p-
menthan-3-one (2.24%), menthofurane
(1.26%), trans-isopulegone (1.09%) and limo-
nene (1.4%). The analysis of Z. hispanica es-
sential oil revealed the presence of a high per-
centage of ketone monoterpene with the pule-
gone (78.6%) its dominant compound. The
ketone (3.33%) represents the second class of
chemical oil, followed by the terpene oxide
(1.31%). The monoterpene with 7 compounds,
represent 2.37% of total oil with limonene as
major compound, unlike sesquiterpene
(0.65%), alkene (0.3%), Ether (0.98%) and
monoterpene alcohol are poorly represented
(Table 2).
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Table 1: Chemical composition of Ziziphora hispanica essential oil
Compounds KI % Compounds KI %
α-pinene 939 0.52 (3Z,5E)-1,3,5-undecatriene 1184 0.41
Cyclohexanone-3-
methyl
952 0.24 1-Dodecene 1192 0.36
Sabinene 976 0.11 α-terpineol 1196 0.71
β-pinene 980 0.5 Puleone 1237 78.6
β-myrcene 987 0.3 Piperitenone 1245 2.9
Isolimonene-trans 983 0.1 8-hydroxy-p-menthan-3-one 1256 2.24
Limonene 1031 1.4 1,3-Dimethyl pyrogallate 1357 0.98
Iso menthone 1130 0.11 α-copaene 1376 0.2
1,8-Cineole 1033 0.1 β-bourbonene 1417 0.1
(-)-L-Isopulegol 1145 0.1 β-caryophyllene 1425 0.4
Camphor 1146 0.06 γ-cadinene 1514 0.1
Trans-isopulegone 1157 1.09 Mint furanone-2 1520 0.59
Menthofuran DB5-785 1164 1.26 Caryophyllene oxide 1582 0.11
neo-Menthol 1166 0.05 2-Pentenoic acid, methyl ester,
(E)
1592 0.18
Table 2: Chemical classes and dominant compound oil from Ziziphora hispanica
Chemical class Nb % Dominant compound %
monoterpene 7 2.59 Limonene 1.06
terpene oxide 2 1.31 Menthofurane 1.26
monoterpene ketone 4 81.45 Pulegone 78.6
monoterpene alcohol 3 0.79 α-terpeneol 0.71
ketone 3 3.07 8-Hydroxy-.delta.-4(5)-p-menthen-3-one 2.24
ether 1 0.98 Syringol 0.98
alkan 2 0.77 (3Z, 5E)-1, 3, 5-undecatriene 0.41
alkene 1 0.30 3-nanone 0.30
sesquiterpene 5 0.91 β-caryophyllene 0.40
others 1 0.18 2-Pentenoic acid, methyl ester, (E)- 0.18
The present research showed that sensitivity
of bacteria Gram-positive, to the essential oil of
Z. hispanica, is higher than that of Gram-
negative bacteria. Antimicrobial activity results
are shown in table 3. The essential oil of aerial
parts of Z. hispanica has a broad spectrum of
antimicrobial activity.
Although this essential oil has remarkably
inhibited the growth of all tested bacteria
including medically important pathogen
Staphylococcus aureus ATCC 6538/P
(inhibition zone is 40 mm). Essential oil has
weakly inhibited the growth of Aspergelus
flavus LBVM20 and A. niger LBBM62, while
its action on Candida albicans ATCC 24433 is
highly active. Anti-bacterial activities of Z.
hispanica essential oil show the presence of
Pulegone found as 77.53% in volatile oil and
also Limonene and Piperitenone can be
responsible in the activity.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Table 3: Antibacterial activity of Ziziphora hispanica essential oil
Strains Inhibition zone
(mm)
Cont
rol
[C] v/v 1 ½ ¼
Bacteria
Bacillus subtilis ATCC 6633 22 22 20 24
Citrobacter sp. 20 20 18 22
Escherichia coli ATCC 25922 22 24 25 28
Enterobacter sp. 22 24 25 20
Klebsiella pneumoniae CIP 53-153 45 49 32 22
Pseudomonas aeruginosa ATCC 27853 38 42 44 26
Pseudomonas syringae pv. Tomato ATCC 1086 35 37 33 20
Staphylococcus aureus ATCC 25923 40 40 41 25
Salmonella enterica CIP 60-62T 45 43 40 30
Fungi
Aspergelus flavus LBVM20 8 8 12 25
Aspergilus niger LBBM62 9 8 8 20
Candida albicans ATCC 24433 33 34 35 22 Inhibition zone (diameter of the disk, 6 mm, included), values represent average of 3determinations;
Control: Chloramphenicol for Bacteria and Amphotericin B for fungi (10 μg/disk);
CIP: Collection of Pasteur Institute, Algeria; ATCC: American Type Culture Collection;
LBM: Laboratory of Biotechnology and Metagenomic, M’sila, Algeria
DISCUSSION
The result of this research is in accordance
with other earlier studies on Ziziphora species
that are all found to be rich in pulegone and
the review of the published literatures reveal
that the composition of Ziziphora species oil
shows large similarity in the major
components, but relative concentrations have
some difference (Gözde et al., 2006; Sonboli
et al., 2006; Ozturk and Ercisli, 2006, 2007;
Aghajani et al., 2008; Amiri, 2009; Maya,
2011; Ozturk et al., 2007 and Soltani, 2012).
The previous studies showed that
Pulegone and Limonene are anti-bacterial
(Maya, 2011). The results in this study are
consistent with the other antibacterial study
results of Ziziphora species and other
pulegone rich plants. However, it has been
reported that the essential oils of pulegone rich
plants such as Micromeria silicica and Mentha
suaveolens inhibited Candida albicans (Gözde
et al., 2006).
The essential oil of Z. clinopodioides
showed good activity against all test bacteria
(Soltani, 2012). The antibacterial activity of
the oil may be associated with the relatively
high pulegone, piperitenone and 1- 8-cineole
content. It has been reported that these
components have significant antimicrobial
activities (Sezik et al., 1991; Meral et al.,
2002; Bakkali and Averbeck, 2008; Sonboli et
al., 2006).
CONCLUSION
In conclusion, the essential oil of the aerial
parts of Z. Hispanica, remarkably inhibited the
growth of all tested gram positive and gram
negative bacteria and the fungus tested. The
essential oil with a composition of pulegone
(77.35%), piperitenone (2.90%) and limonene
(1.06%) and its observed antibacterial
properties show that the essential oil could be
evaluated in the pharmaceutical industry as a
possible new pulegone resource.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 73–80
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ACKNOWLEDGEMENTS
This work was supported in part by the
Laboratory of the Chemistry of Heterocycles,
Blaise Pascal University (France) and MESRS
of Algeria.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
COMPARISON OF COLCHICINE CONTENT BETWEEN
HYSTERANTHOUS AND SYNANTHOUS COLCHICUM SPECIES IN
DIFFERENT SEASONS
Alirezaie Noghondar Morteza1*, Arouee Hossein
2, Shoor Mahmoud
2, and Rezazadeh
Shamsali 3
1*
PhD student, Ferdowsi University of Mashhad, Agriculture Faculty, Horticultural Sciences Department,
Mashhad, Iran 2 Assistant Professor, Ferdowsi University of Mashhad, Agriculture Faculty, Horticultural Sciences
Department, Mashhad, Iran 3 Assistant Professor, Institute of Medicinal Plants, Department of Pharmacognosy and Pharmaceutics,
ACECR, Tehran, Iran
*Corresponding author: Email: [email protected]
Received: 13/12/2012; Revised: 24/01/2013; Accepted: 30/01/2013
ABSTRACT
In order to compare of different phonological stages and seasonal changes of colchicine content
between hysteranthous and synanthous colchicum species, amount of colchicine was determined in
Colchicum speciosum Steven, C. kotschyi Bioss and C. robustum Stefanov, in different seasons,
2009–2010. The observations under wild conditions showed, that the leaves of appeared with flowers
in the same stage of life cycle (synanthous) in C. robustum, while in case of C. kotschyi and C.
speciosum flowers occurred first and leaves later, in another developmental stage (hysteranthous).
Seed’s colchicine content in C. robustum, C. kotschyi and C. speciosum was obtained as 1.28, 0.46
and 0.92 mg g-1
dry weight, respectively. Corm’s colchicine content was higher in C. speciosum than
the other species in all seasons. The highest colchicine content of corm in C. speciosum was obtained
in winter and autumn (2.17 and 2.13 mg g-1
dry weight, respectively), while in C. robustum and C.
kotschyi it was found in autumn, 0.49 and 0.77 mg g-1
dry weight, respectively. The lowest
colchicine content of corms was obtained in summer, when the corms were dormant before
flowering stage, in C. speciosum and C. kotschyi, 0.131 and 0.0058 mg g-1
dry weight, respectively,
whilst in C. robustum obtained in winter, 0.08 mg g-1
dry weight, synchronous to flowering and
vegetative growth.
KEYWORDS: Colchicine content, Colchicum kotschyi, C. speciosum, C. robustum, Flowering
stage, Hysteranthous, Root activity, Synanthous, Seasonal changes.
Research article
Cite this article:
Alirezaie Noghondar Morteza, Arouee Hossein, Shoor Mahmoud, and Rezazadeh Shamsali (2013),
COMPARISON OF COLCHICINE CONTENT BETWEEN HYSTERANTHOUS AND
SYNANTHOUS COLCHICUM SPECIES IN DIFFERENT SEASONS., Global J Res. Med. Plants &
Indigen. Med., Volume 2(2): 81–88
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 81–88
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
The genus Colchicum belongs to the family
Colchicaceae, which comprises of 19 genera,
and 225 species (Nordenstam, 1998). Plants of
the genus Colchicum have been known for more
than 2000 years for their marked beneficial and
poisonous effects (Brickell, 1984). The modern
medicine uses Colchicum as a source of
therapeutically active alkaloids called
colchicinoids. One of the most abundant
alkaloid - colchicine, is known to have
cancerostatic, antirheumatic, antimitotic,
antiinflammatory, cathartic and emetic effects. It
is also applied in plant breeding to induce
polyploids (Komjatayova et al., 2000; Frankova
et al., 2005). In addition to the genus
Colchicum, colchicine was reported from
species belongs to Merendera and Gloriosa
genera, which belonging to the Colchicaceae
family (Nordenstam, 1998).
Many factors are interfering in biosynthesis
of secondary metabolites such as essential oils
and alkaloids. The study conducted by Takia et
al. (2013), has shown that essential oil
composition and content in four populations of
Pituranthos scoparius were different. Very little
is known about the factors interfering with the
biosynthesis of colchicine-like alkaloids. Results
obtained by Sütlüpinar et al. (1988), indicated
that the composition of tropolone alkaloids
differs in different parts of the plants and varies
during the different growth stages (Sütlüpinar et
al., 1988). Presence and concentration of
colchicine is determined by a variety of
environmental factors including season (Vicar et
al., 1993; Poutaraud and Girardin 2002; Alali et
al., 2006) and resource availability (Hayashi et
al., 1988; Pouraraud and & Girardin, 2005;
Mróz, 2008) as well as genetic variations
between populations and individuals (Poutaraud
and Champay, 1995). Also colchicine content
varies among different organs of the plant body
(Sütlüpinar et al., 1988; Alali et al., 2004; Alali
et al., 2006).
Among all species of Colchicum, C.
autumnale is the best source for colchicine. The
richest plant parts in colchicine are the corms
and seeds. C. autumnale seeds contain 0.6-1.2%,
while corms contain up to about 0.6%. Seeds are
mainly used by the pharmaceutical industry for
the extraction of colchicinoids (Trease & Evans,
1983). The content of colchicine alkaloid in
corms, stems, leaves, and flowers of C.
cilicicum were 0.05%, 0.01%, 0.01% and 0.20%
(g% dry weight), respectively (Sütlüpinar et al.,
1988). In another study by Alali et al. (2004), C.
stevenii corms, flowers and leaves were reported
to contain 0.17, 0.12 and 0.20 (wt/wt) g%,
respectively, while C. hierosolymitanum corms
and flowers were found to contain 0.13 and 0.09
(wt/wt) g%, respectively. Ondra et al. (1995),
assayed corms of seven Turkish Colchicum
species; namely: C. macrophyllum, C. turcicum,
C. cilicicum, C. kotschyi, C. bornmuelleri, C.
speciosum and C. triphyllum for their
colchicinoid alkaloids. Colchicine content was
found to be 222.3, 323, 300, 1058, 3063, 4245
and 958 µg g-1
dried drug, respectively.
Colchicine variation in different organs of
plant and during different growth stages has
been studied by researchers. Colchicine and
demecolcine were determined in raw and dried
leaves, stems, mother and daughter corms of C.
autumnale in four stages of its ontogenesis by
Vicar et al. (1993). They found that colchicine
content in raw material varies during plant
growth. Colchicine content in C. brachyphyllum
and C. tunicatum, was determined during
different growth stages by Alali et al. (2006).
Underground parts in both species and during
different growth stages, always showed higher
colchicine content than the above ground parts.
In C. brachyphyllum, total colchicine content of
underground parts during flowering stage was
found to be about 0.15% (wt/wt), while that of
aerial parts was only about 0.04% (wt/wt). In C.
tunicatum, total colchicine content of
underground parts was found to be 0.12%
(wt/wt) and 0.13% (wt/wt) during flowering and
vegetating stages, respectively, while that of
aerial parts was only about 0.04% (wt/wt) and
0.02% (wt/wt), respectively (Alali et al., 2006).
Generally, geophytes are plants that survive
by subterranean storage organ with renewal
buds (Raunkiaer, 1934). They divide into two
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groups – synanthous and hysteranthous one. The
leaves of synanthous geophytes coexist with
flowers in the same stage of life cycle. In case of
hysteranthous plants flowers occur in the first
and leaves later, in another developmental stage
(Dafni et al., 1981). A special case is the
hysteranthous plant Colchicum tunicatum which
perceive the photoperiodic signal when the dry
bulb lies well below the soil surface (Halevy,
1990). C. speciosum, C. kotschyi Boiss, and C.
robustum stefanov, are three wild growing
Iranian Colchicum species (Presson, 1992). C.
speciosum Steven and C. kotschyi Bioss are
hysteranthous but C. robustum is a synanthous
species (Presson, 1992).
So far no study has been performed on
colchicine content variation between synanthous
and hysteranthous Colchicum species in
different seasons, thus the aim of this study was
to evaluate phenological changes and their
relationship with corm and seed colchicine
content variation among three Iranian native
Colchicum species, under their habitat
conditions.
MATERIAL AND METHODS
Plant Material
The corms of three wild Colchicum species
were collected in different seasons (spring,
summer, autumn and winter during 2009–2010,
and seeds were collected in spring 2010. Corms
and seeds of C. speciosum, C. robustum and C.
kotschyi were collected from Khalkhal-Asalem
road, Ardabil province, at an altitude of 940m in
Iran, Babaaman Mountain, North Khorassan
Province, at an altitude of 1091m in Iran and
Noghondar valley near Mashhad, Razavi
Khorasan province, at an altitude of 1400m in
Iran, respectively. The collected materials of
three species were identified by Mohammad
Reza Joharchi, Ferdowsi University of Mashhad
Herbarium (FUMH). Voucher specimens of C.
kotschyi (Herbarium Number: 39516), C.
robustum (Herbarium Number: 39519) and C.
Speciosum (Herbarium Number: 39531) were
registered. These are kept in the herbarium of
FUMH.
Recording developmental stages
To study the plant phenology in wild
conditions observations were carried out for
three species from three different locations
during 2009–2010. Observations were including
of developmental stages such as beginning of
flowering, peak flowering time, root formation
time, beginning of vegetative growth, fruiting
and capsule formation and daughter corm
formation in wild conditions.
Extraction and Isolation
The methods described by Rosso and
Zuccaro (1998) and Alali et al. (2006), were
adopted with some modifications. Acetonitrile,
methanol and other reagents were of
chromatographic grade and prepared from
Panreac (Spain). Reference standard of
colchicine was prepared from USP.
The corms were sliced into small pieces and
air-dried at room temperature together with the
seeds. After drying, exact weight of 2 g of
corms (collected in different seasons) and 2 g of
seeds of three Colchicum species were grounded
to powder in a laboratory mill and then used for
extraction. Powdered material placed into
250 mL Erlenmeyer flasks and extracted with
100 mL of methanol in 35oC for 1h with
ultrasonic apparatus. Afterwards, plant residues
were filtered through Wattman filter paper and
the filtrates were saved. Then plant residue was
transferred into Erlenmeyer flasks again and
extracted with 50mL of methanol in 35oC for
30min with ultrasonic apparatus and then
filtered. Plant residues were washed with 10 mL
of methanol and then filtered. The collected
filtrates and washes were combined and
transferred into a 250 mL separatory funnel and
extracted with petroleum ether (30 mL × 3) with
frequent shaking for 30 min in order to remove
non-alkaloid substances. 10 mL of distillate
water was added each time for better separation
and creation of two separate phases. The
resulting methanolic phase was transferred to an
empty separatory funnel and extracted with
chloroform (30 mL × 3) for 10 min. The
chloroform phases obtained from three stages
were collected and Sodium sulphate Anhydrous
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was added to the chloroformic solution for
dewatering of it and then filtered through filter
paper. The chloroformic extract was dried in
vacuum and then dissolved in 5 ml of HPLC
grade methanol and injected to the HPLC
instrument. Injection volume at 50 μl, room
temperature, detection at 243 nm. All analyses
were done in duplicate.
HPLC instrument was Knauer ® (Germany)
equipped with auto sampler and column was
Bondapak C18 (Technochrom) micrometer
particles and 4.9 mm id and 250 mm in length.
An UV detector K-2501 and a dynamic mixing
chamber were employed. Mobile phase system
consisted from phosphate buffer pH=6 and
acetonitrile (77: 23). For preparation of
phosphate buffer 800 mg of NaH2Po4 and
200mg of Na2HPo4 were dissolved in 1000mL
of HPLC grade water and the pH was adjusted
on 6. The flow rate was adjusted to 2mL/min
and detection was performed at a wavelength of
243nm. The stock solution of colchicine
standard was prepared by accurately weighing
of colchicine reference standard and then diluted
using HPLC grade methanol to construct
calibration curve of six –points (30, 50, 75, 90,
100 and 120 ppm). Figure 1 shows colchicine
HPLC analysis standard curve.
Figure. 1. Colchicine HPLC analysis standard curve
RESULTS AND DISCUSSION
Developmental stages
Table 1 shows, beginning time of
developmental stages in three colchicum
species under their habitat conditions. The
results showed that flowering started sooner in
C. Speciosum (end of August) and C. kotschyi
(middle of September) than to C. robustum
(end of January). Fruiting and capsule
formation started later in C. robustum (middle
of April) than to C. speciosum (beginning of
April) and C. kotschyi (end of March). In all
species root activity got initiated in middle of
autumn (Table 1).
Observations showed that C. kotschyi and
C. speciosum were hysteranthous geophyte
(flowers develop first and leaves later) and
autumn-flowering species but C. robustum was
a synanthous geophyte (leaves coexist with
flowers in the same stage of the life cycle) and
winter-flowering species. This type of
obviously hereditary phenological behaviour is
rather the rule in the genus Colchicum, in
contrast to the onset of leaf growth which
seems to be largely environmentally triggered
(Burtt, 1970; Gutterman and Boeken, 1988;
Persson, 1999).
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Table 1. Beginning of developmental stages of three colchicum species under natural conditions
Different
Species
Beginning of
flowering
Pick time of
flowering
Root
formation
Vegetative
growth
Fruiting and
capsule
formation
Daughter
corm
formation
C. speciosum Ea-Aug E-Sep E-Oct E-Mar B-Apr E-May
C. kotschyi M-Sep B-Oct B-Nov B-Feb E-Mar M-May
C. robustum E-Jan M-Feb B-Dec B-Feb M-Apr B-May
Notes: a B, M and E indicate the beginning, middle and end of each month, respectively
Colchicine content
The results of HPLC analysis of plant
extracts are summarized in table 2. The level of
colchicine varies in different seasons as well as
species and plant parts. Seed’s colchicine
content in C. robustum was higher than the
other species. Seed’s colchicine content in C.
speciosum, C. kotschyi and C. robustum was
0.92, 0.46 and 1.28 mg g-1
dry weight (DW),
respectively (table 2). The amounts of corm
colchicine in C. speciosum were higher than the
other species in all seasons. Among different
seasons the highest colchicine content of corm
in C. speciosum was obtained in winter
(2.17 mg g-1
DW), while in C. robustum and C.
kotschyi it was found in autumn, 0.49 and
0.77 mg g-1
DW, respectively. The lowest
colchicine content of corm was obtained in
summer in C. speciosum and C. kotschyi was
found to be about 1.31 and 0.058, respectively,
while in C. robustum it was in winter, 0.08 mg
g-1
DW.
Corm’s colchicine content in C. speciosum
and C. kotschyi (as hysteranthous species) in
autumn and winter were higher than to spring
and summer, while in C. robustum (as a
synanthous species) the highest corm
colchicine content was obtained in autumn. The
lowest colchicine content in C. kotschyi and C.
speciosum was obtained in summer, whilst in
C. robustum, it was observed in winter.
Colchicine content in different species
varies considerably during different seasons.
Matching of the table related to developmental
stages with seasonal variation of colchicine
content indicates that corm colchicine content
in the three colchicum species studied was high
in autumn (at the time of root activity). The
lowest colchicine content of corm in C.
speciosum and C. kotschyi (as hysteranthous
species) was observed when the corms were
dormant, while in C. robustum (as a synanthous
species) it was obtained during flowering and
vegetative stages. During flowering stage and
in the absence of leaves, the only source of
colchicine in flowers could be due to the
translocation of colchicine from corms and this
may explain the slightly low corm colchicine
content at flowering stage (Al-Fayyad et al.,
2002).
Seed colchicine content in C. robustum was
higher than those of the other species.
Previously reported that the amount of seed
alkaloid and colchicine content is more in
unripe seed and declines as the seeds mature
(Poutaraud and Girardin, 2003; Alali et al.,
2006). Since In the present study, the capsules
and seeds of C. speciosum and C. kotschyi were
formed sooner than those of C. robustum so it
seems that, less colchicine content in C.
speciosum and C. kotschyi seeds had been due
to more mature their seeds than those of C.
robustum.
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Table 2. Mean Colchicine content of different organs of the three Colchicum species (mg g-1
dry weight)
during different seasons.
Notes: a Colchicine content is expressed as mass of colchicine in 1 gram dry weight ± standard deviation, derived from
the average of two extraction replicates, each run in duplicate
Alkaloids are responsible for the plant
adaptation to its environment. It is known that
alkaloids are efficiently used as defensive
agents and they may be moved around the plant
to those parts needing greater protection during
growth and development (Harborne, 1997). As
part of their defences against herbivores, many
geophytes are toxic and unpalatable, or have
developed different physical defences against
herbivores (Lovegrove & Jarvis, 1986;
Go´mez-Garcı´a et al., 2004). This is the case
of different plant species of colchicum, which
contain colchicine.
CONCLUSION
In conclusion, what the results suggest is
that the highest corm colchicine content in the
three species was found in autumn (the period
of root activity). Thus the corms of these three
species are better to be collected in autumn
from their local habitats, to ensure that
maximum of colchicine is achieved. The lowest
corm colchicine content in C. robustum (as a
synanthous species) was observed in winter (at
flowering stage) whereas in C. speciosum and
C. kotschyi (as hysteranthous species) in
summer, when the corms are dormant.
However, more species of colchicum need to be
examined to determine a stronger relationship
between developmental habit (specially
flowering habit) and colchicine content.
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Source of Support: Nil Conflict of Interest: None Declared
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
ECOLOGICAL AND MEDICINAL INTEREST OF TAZA NATIONAL PARK
FLORA (JIJEL - ALGERIA)
BOUNAR Rabah1,2
*, REBBAS Khellaf2, GHARZOULI Rachid
1, DJELLOULI Yamna
3and
ABBAD Abdelaziz4
1 Department of Biological Sciences, University of Ferhat Abbas Setif 19000, Algeria.
2 Department of Nature and Life Sciences, University of M'Sila 28000, Algeria.
3 Department of Geography, University of Maine, 72085 Le Mans, France.
4 Faculty of Sciences, University Cadi Ayyad, Semlalia, BP 2390, Marrakech, Morocco.
*Corresponding Author: E-mail: [email protected]
Received: 08/01/2013; Revised: 26/01/2013; Accepted: 29/01/2013
ABSTRACT
The forest of Taza National Park (NP), located in North-Eastern Algeria, is characterized by a
high floristic diversity. Analysis of the park flora showed 420 species belonging to 258 genera and
71 botanical families. Asteraceae (54 species), Fabaceae (37), Poaceae (34), Lamiaceae (26) and
Brassicaceae (24) are the most dominant families. The endemism rate is around 12.38% (52
species); approximately 21% of endemic species of Algeria. Rare and very rare species were
estimated to be 120 taxa representing 28.57% compared to the park flora. Analysis of global
phytochoric spectrum shows dominance of native Mediterranean species (193 species). This floristic
wealth contains a number of 205 species of medicinal interest.
KEYWORDS: Floristic diversity, medicinal plants, Taza National Park, Algeria.
Research article
Cite this article:
BOUNAR R, REBBAS K, GHARZOULI R, DJELLOULI Y and ABBAD A (2013),
ECOLOGICAL AND MEDICINAL INTEREST OF TAZA NATIONAL PARK FLORA (JIJEL -
ALGERIA), Global J Res. Med. Plants & Indigen. Med., Volume 2(2): 89–101
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 89–101
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INTRODUCTION
Algeria, like all Mediterranean countries,
has long been involved in the politics of
preservation and conservation of biodiversity
through the creation of several National Park’s.
Currently, it counts eight NP’s including all
original landscapes the main hot spots of plant
biodiversity in the country (Benhouhou & Vela,
2007). Several research works mainly focused
on the identification and mapping of the phyto-
biodiversity have been made in these hot spots:
NP of Chrea (Zeraia, 1981), NP of El Kala
(Stevenson et al., 1988; Belouahem et al.,
2009), NP of Tlemcen (Yahi et al., 2007;
Letreuch-Belarouci et al., 2009) and NP of
Gouraya (Rebbas, 2002; Rebbas et al., 2011).
These research works underlined the rich
flora of these areas and highlighted panoply of
endemic and/or rare species which must be
placed in conservation priorities. This work
also evoked the advanced state of degradation
of these natural ecosystems and emphasized the
importance of such an inventory list in the
rational management of these natural
ecosystems. Indeed, several authors evoked that
the conservation and the development of a
natural ecosystem pass by a good knowledge of
its biodiversity (Daget & Poissonnet, 1971;
Médail & Quezel, 1997; Véla & Benhouhou,
2007).
In order to know the vascular flora of these
natural environments, we are interested by the
study the floristic diversity of one of the most
original ecosystems, at a biogeographic and
ecological level, of the Algerian North-eastern
sector. It is about Taza NP which belongs to the
small Kabylia sector of the Babors (Figure 1)
and is regarded as the most wooden area in
Algeria with a very high rate (Bensettiti &
Abdelkrim, 1990).
This work fills the gaps on the state of
current knowledge on the vascular flora of the
Taza National Park. Indeed, the only known
floristic inventory work known in the area and
concerned neighborhoods of the Park primarily
(Gharzouli 1989; Gharzouli & Djellouli, 2005
Gharzouli, 2007, Bounar, 2003). Only work of
floristic synthesis which refers to all North
Eastern Algeria remains very old and not
updated (Khelifi 1987; Aouedi 1989; Aktouche
et al., 1991). Other research made on some
forest formations of the park remains very
sketchy. As examples, we can mention
phytosociological work of Zeraia (1981),
Dahmani (1984) and Bensettiti & Abdelkrim
(1990). Knowledge of the diversity of species
of medicinal interest of this area allows us to
offer solutions for conservation and recovery of
these resources within the framework of
sustainable development.
I- Presentation of the study area
Taza NP was created in 1984 on a total area
of 3807 ha. It is located in the North-East of
Algeria between geographical coordinates 36°
35'–36° 48' North latitude and 5° 29'–5° 40'
West longitude. Taking part of the small
Kabylia of Babors, it opens onto the
Mediterranean Sea in the Gulf of Bejaia (Figure
1). According to the rainfall map established by
the National Agency for Water Resources
(NAWR, 1996), the study area is situated in
annual sections ranging from 850 mm–
1750 mm. Average minimum temperature of
the coldest month (January) varies between
6.1° C and 8.1° C. Maximum temperatures of
the hottest month (July) is between 30.2° C and
34.8° C. Dry period varies from 3–5 months.
High relative humidity of the air (80%)
promotes the installation and maintenance of
quite important plant diversity. Emberger
pluviothermic quotient Q2 (Emberger, 1955)
varies between 110 and 124 placing the Park in
humid bioclimatic stages to sub-humid with
variations to mild and warm winter (Daget &
David, 1982).
The Park presents a very rugged terrain
including several mountain ranges oriented
from east to west with altitude varying from
480 m to the highest point in the area (1121 m).
These orographic elements give a general
configuration in folds in North-eastern and
South-western orientations. Geologically, the
area is dominated by sedimentary grounds of
sandstone and volcanic soils in North zones
(Obert, 1970).
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Figure 1 Localization map of the study area
These climatic and lithological
characteristics determine a rich and diversified
flora whose principal forest species are the zeen
oak (Quercus canariensis Willd) , which covers
more than 40%, the cork oak (Quercus suber L)
with 39% and afares oak (Quercus afares
Pomel) with only 5% (Bensettiti & Abdelkrim,
1990). According to Maire (1926), Quezel &
Santa (1962-1963), Zeraia (1983), Barry et al.
(1974), Quezel (1978) and Barbero et al.
(2001), Taza NP is on the phyto-geographical
region Mediterranean, North African
Mediterranean area and belonging to the
Numidian.
II - METHODOLOGY
Park flora was established by floristic
surveys carried out, according to the
phytosociological method, in different types of
vegetation. Surfaces floristically homogeneous
were defined on the basis of most common
ecological parameters such as altitude,
exposure and slope. Covering of the vegetation,
by layer, was also taken into account. 63
floristic surveys were carried out. Survey
surface varies according to vegetation types. It
oscillates between 300–400 m² for forest
vegetation and between 5 and 10 m² for
rupicolous vegetation. Surveys were conducted
during years 2005 and 2008.
The floristic surveys were carried out
according to a subjective sampling in all
vegetation types of the Park. Samples of plant
species collected were determined in laboratory
using different flora: Maire (1952-1987),
Quezel & Santa (1962-1963), Fennane et al.
(1999; 2007) and Valdes et al. (2002). Species
nomenclature adopted was according to "Med-
Cheklist, critical inventory of vascular plants of
circum Mediterranean countries"(Greuter et al.,
1984).
Control samples of collected species were
deposited in the laboratory of Setif University.
Chorologic types of various identified taxa
were assigned as indicated in consulted floras;
special attention was given to endemic and/or
rare species. Analysis of the floral study area
and various ethnobotanical fieldwork in the
Park surrounding regions, allowed us to have
an extensive list of medicinal plants used by the
neighboring population.
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III-RESULTS AND DISCUSSION
Specific richness
Enumerated taxa were 420 species and
subspecies belonging to 258 genera and 71
botanical families of vascular plants
(phanerogams and vascular cryptogams);
approximately 10% of the Algerian total flora
estimated at 3139 species (Quezel & Santa,
1962; 1963). Phanerophytes (41 species)
occupy 9% of the Park flora. On the total flora
recorded at the Park, Asteraceae, Fabaceae,
Poaceae, Lamiaceae, Brassicaceae,
Caryophyllaceae and Rosaceae were best
represented with more than 20 species each.
These families represent nearly 40% of the total
richness of the Park.
Our results are consistent with those of
Gharzouli & Djellouli (2005). This wealth
places the Park among the most diversified
ecosystems in the country, as is the case for all
Small Kabylia (Gharzouli, 2007; Vela &
Benhouhou, 2007). This floristic wealth of the
Park is probably due to (i) its geographical
position opening directly on the Mediterranean
Sea and therefore exposed to the maritime
influences of the North-West (ii) diversity of
habitats resulting from climatic and edaphic
heterogeneity and (iii) a relatively weaker
exploitation of the medium compared to other
ecosystems.
Chorological Type
Floristic analysis shows the presence of
several phytochoric units (Figure 2).
Mediterranean one is the most representative
with 193 species. This situation is common to
most natural ecosystems of Algeria (Quezel,
1964; 2002) and the Mediterranean basin
(Dahmani, 1984; Quezel & Barbero, 1990;
Quezel & Medail, 2003). This whole
Mediterranean is divided into several subsets:
s.l. Mediterranean (114 species), western
Mediterranean (42 species), Ibero-
mediterranean (20 species), oro-mediterranean
(8 species), central mediterranean (2 species)
and eastern mediterranean (7 species). Northern
chorologic species (Nordic) are relatively well
represented in the Park, such as those of
european element (20 species), eurasian (41
species), paleo-tempered (22 species), circum-
boreal (6 species), oro-european (01 species)
and atlantic (14 species). Other species
correspond to transition elements between
chorological mediterranean and those
neighbors such as the euro-Mediterranean (30
species), mediterranean-irano-turanian (6
species), macaronesian, mediterranean and
asian mediterranean with 4 species each.
Figure 2 Chorological spectrum of Taza National Park
46%
12%
21%
1%20%
Mediterranean
Endemic
Nordic
Paleotropical
Wide distribution
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Analysis of endemism
52 taxa were recorded, about 12.38% of
total species of the Park and 9.47% compared
to the total endemic flora of the country
estimated at 549 species (Quezel, 1964) and
nearly 12.7% of northern Algeria (Vela &
Benhouhou, 2007). Endemism rate is relatively
high compared to that recorded in several Parks
in central and eastern Algeria such that of
Belezma -Batna- (32 species), Gouraya -
Bejaia- (26 species) (Rebbas, 2002; Rebbas et
al., 2011), Djurdjura (35 species) (Meribai,
2006) and Kala -Taref- (75 species)
(Stevenson, 1988).
Endemic flora of Taza Park consists mainly
of endemic Algerian species (18 species),
North Africa (22 species), Algerian-Moroccan
(5 species), Algerian-Tunisian (7 species).
13.47% of the Park endemic taxa belonged to
Asteraceae and Lamiaceae families with 7
species each.
Analysis of the rarity
Relying on Quezel & Santa data (1962;
1963) nearly 120 species were reported as rare
or very rare. On the basis of these data, the
Taza NP records a 28% rarity rate of all its
inventoried taxa and around 7% compared to
rare species of northern Algeria and about 6.6%
over the entire national territory. Compared to
the phyto-geographical of Kabylia totaling
approximately 487 rare species (Vela &
Benhouhou, 2007), Taza NP occupies nearly
24.6% (Figure 3).
Among the 129 Algerian taxa Red listed by the
International Union for Nature Conservation
(1980), 12 species belong to the Taza NP
spread over the studied three types of
formations (Tables 1 and 2).
Figure 3: Rare Plants in Taza National Park (Photos: K. Rebbas, 2011)
1. Phlomis bovei de Noé
2. Berberis hispanica Boiss. et Reut.,
3. Atropa belladonna L.
4. Crataegus laciniata Ucria.
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Table 1 : Number of rare and endemic species per botanical family
Botanical
families Number of
endemic species
Percentage
(%)
Number of rare
species
Percentage
(%)
Asteraceae 07 13.46 16 13.33
Lamiaceae 07 13.46 07 5.83
Poaceae 03 5.76 11 9.16
Caryophyllaceae 03 5.76 09 7.5
Brassicaceae 03 5.76 12 10
Fabaceae 03 5.76 13 10.83
Scrofulariaceae 03 5.76 04 3.33
Apiaceae 03 5.76 08 6.66
Ranunculaceae 02 3.84 05 4.16
Crassulaceae 02 3.84 03 2.5
Campanulaceae 02 3.84 01 0.83
Pinaceae 01 1.92 01 0.83
Fagaceae 01 1.92 - -
Berberidaceae 01 1.92 02 1.66
Geraniaceae 01 1.92 02 1.66
Thymelaeaecea 01 1.92 02 1.66
Violaceae 01 1.92 01 0.83
Cistaceae 01 1.92 - -
Primulaceae 01 1.92 01 0.83
Convolvulaceae 01 1.92 02 1.66
Plantaginaceae 01 1.92 - -
Rubiaceae 01 1.92 04 3.33
Caprifoliaceae 01 1.92 04 3.33
Valerianaceae 01 1.92 02 1.66
Linaceae 01 1.92 - -
Rosaceae - - 07 5.83
Saxifragaceae - - 03 2.59
Total 52 120 100
Medicinal plants
205 species of medicinal interest were
enumerated. Development of research in field
of pharmacology and identification of species
active principles will create economic activity
in use of plants organized in a friendly
safeguard flora.
As in the majority of Algerian areas, some
of these species are employed by inhabitants
bordering the Park as traditional medicine and
are marketed by herbalists (Alnus glutinosa L.,
Arbutus unedo L., Asphodelus microcarpus
Salzm. & Viv., Asparagus officinalis L.,
Clematis flammula L., Ceterach officinarum
Lamk, Crataegus laevigata (Poiret) DC,
Crataegus laciniata Ucria, Mentha pulegium
L., Mentha spicata L., Inula viscosa L., Mentha
rotundifolia L., Myrtus communis L., Opuntia
ficus indica (L.) Mill., Ficus carica L., Pistacia
lentiscus L., Prunus avium L., Punica
granatum L., Quercus suber L., Juniperus
oxycedrus L., Nerium oleander L., Teucrium
polium L., Thapsia garganica L., Ulmus
campestris L).
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Table 2: Rate rarity by chorological origin
Chorological origin Total
number
of species
Percentage
rate (%)
Degree of rarity
Total species rare
and very rare
Percentage
rate (%)
Mediterraneans 193 45.95 73 37.82
Mediterranean 114
Western Mediterranean 42
Ibero-Mauritanian 20
Euro-Mediterranean 08
Central Mediterranean 02
East Mediterranean 07
endemics 52 12.38 11 21.15
Algerian endemic 18
North African 22
Algerian-Moroccan 05
Algerian-Tunisian 07
Nordics 90 21.42 19 21.11
Eurasiatic 41
European 20
Paleo-Temperate 22
Circum-Boreal 06
Oro-European 01
paleotropicals 02 0.47 1 50
Wide distribution 83 19.78 16 19.27
Euro-Mediterranean 30
Atlantic-Mediterranean 14
Macaronesian-
Mediterranean
04
Eurasiatic-
Mediterranean
02
Asiatic-Mediterranean 04
Irano-Turanian-
Mediterranean
06
Eurasian-Macaronesian 03
Mediterraneo-Saharan-
Arabian
02
diverse 18
Total 420 100 120
Many plants were subject (of) to
phytochemical analysis and ethnobotanical
studies in North Africa in general and in
Algeria in particular. Majority of these plants
appear in the floristic list of the study area like:
Berberis hispanica Boiss. & Reut., Bupleurum
montanum Coss, Cynodon dactylon L., Inula
crithmoides L., Inula viscosa L., Origanum
glandulosum Desf., Olea europaea L., Pistacia
lentiscus L., Phlomis bovei de Noé, Salvia
verbenaca L., Teucrium polium L. Ricinus
communis L (Chemli, 1997; Hmamouchi, 1997;
Baba Aissa, 1999; Ruberto et al., 2002;
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Belarchaoui & Boukhadra, 2006; Boulaacheb,
2006; Sari et al., 2006; Hseini & Kahouadji,
2007; Liolios et al., 2007; Benguerba, 2008;
Laouer et al., 2009; Hachicha et al., 2009 ;
Derridj et al., 2009; Ouled Dhaou et al., 2010;
Cahuzac-Picaud, 2010; Makhlouf et al., 2010;
El Youbi, 2011; Rebbas et al., 2012; Lemoui et
al., 2012; Sari et al., 2012; Hendel et al., 2012).
The anarchy in exploitation of the species
known for their therapeutic virtues constitutes a
risk for their survival. Certain species are in
danger of extinction because of their
overexploitation (abusive pulling up). It is the
case of Lamiaceae species which are uprooted
(torn off with their roots), to be sold in towns
and villages of the area, as: Teucrium polium
L., Mentha rotundiflolia L., Origanum
glandulosum Desf
CONCLUSION
Analysis of the floristic diversity of Taza
NP shows well its great richness and its
ecological and phytogenetic originality. These
data justify its classification with all small
Kabylia as a hot spot in northern Algeria (Vela
& Benhouhou, 2007). Despite legislative
protection, this Park, like most Mediterranean
natural ecosystems, is subject to a worrying
degradation. Indeed, human activities (anarchic
collection of wood, cork exploitation, uprooting
plants of interest) and uncontrolled pasture are
seriously detrimental to the richness. To face
these problems and to keep the ecological
integrity of the Park, an integrated strategy for
conservation of biodiversity must be installed.
This strategy must be focused primarily on
tree forestation of the Park, especially with
zeen oak (Quercus canariensis Willd), cork oak
(Quecus suber L) and afares oak (Quercus
afares Pomel) which constitute the essential
structure of this natural ecosystem. These
principal forest formations harbor several
endemic and/or rare genera like Cyclamen,
Corydalis. Many rare or endangered species of
the Park deserve to be integrated in the Red
List of the International Union for
Conservation of Nature (IUCN). It is about
Galium odoratum (L) Scop, Satureja juliana
L., Viburnum lantana L., Hieracium ernest
Maire, Convolvulus dryadum Maire, Stellaria
holostea L, Chrysanthemum fontanesii L.,
Bupleurum montanum Coss, Quercus afares
Pomel and Sedum pubescens Vahl. (Table 3).
Table 3 : Rare and endangered species in Taza National Park.
Species not listed in the IUCN Red List Species listed in the IUCN Red List
Galium odoratum (L) Scop Arabis doumetii Coss.
Satureja juliana L. Saxifraga numidica Maire
Hieracium ernest Maire Teucrium kabylicum Batt.
Viburnum lantana L. Fedia sulcata Pomel.
Convolvulus dryadum Maire Carum montanum (Coss & Dur.)Benth.
Stellaria holostea L. Lonicera kabylica Rehder.
Chrysanthemum fontanesii L. Teucrium atratum Pomel.
Bupleurum montanum Coss. Epimedium perralderianum Coss.
Quercus afaresPomel Phlomis bovei de Noé.
Sedum pubescens Vahl. Sedum multiceps Coss & Durieu.
Pimpinella battandieri Chabert
Moehringia stellaroides Coss.
IUCN :International Union for Conservation of Nature
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Increasing ethno botanical studies will
allow a better potential understanding of this
field, evaluate consequent risks to the use of
certain toxic plants and adopt a new
management approach for protection and
preservation of natural resources (Lahsissene et
al., 2010). A large number of spontaneous
species of the study area are used in medicine
and food like fodder. Culture of these species
for economic interest, instead of anarchic
gathering, can improve the income of local
people while ensuring the conservation of plant
diversity (Bounar et al., 2012).
For the extraction of active ingredients, the
creation of plots of medicinal plants selected,
from lists established by floristic inventories,
can replace the one gathered. In Algeria, the
market for plants with medicinal properties is
uncontrolled (Boulaacheb et al., 2006).
Considering the various uses of these plants, a
regulation seems necessary. So every country
must define its own specifications (Veuillot,
2001).
Rare and endemic species of the study area
form a draft list of local red rare and
endangered flora. The protection and
conservation of these formations are needed
more than ever before and should receive strict
protection. Tourism activities and grazing may
be detrimental to the biodiversity of the Park.
Urgent solutions must be found to ensure their
survival.
South of the Mediterranean, where the
situation is much more serious,
accomplishments are sporadic and generally
ineffective. Only authoritative decisions taken
by national leaders would likely aim at
preserving some ecosystems or certain groups
particularly at risk. It is this desire that has been
taken in Rabat in 1987, at the meeting for the
conservation of plant resources in the countries
of North Africa (Quezel et Barbero, 1990).
ACKNOWLEDGEMENTS
We are very much grateful to all the
personnel of Taza National Park; BOUAZID
Tayeb (university of Setif); SARI Madani and
HENDEL Noui from university of M’sila for
their help.
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ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
CONSERVATIVE PRODUCTION OF BIODIESEL FROM WASTE
VEGETABLE OIL
Chethana G.S1*, Reddy K Dayakar
2, Vijayalakshmi
3
1Research Associate, R & D, Sri Sri Ayurveda Trust, 21
st Km from Bangalore, Kanakapura Road,
Bangalore-82, Karnataka, India 2, 3
Department of Biotechnology, Oxford College of Science, HSR Layout, Bangalore, Karnataka, India
*Corresponding Author; Email Id: [email protected]
Received: 08/01/2013; Revised: 11/02/2013; Accepted: 15/02/2013
ABSTRACT
Biodiesel can be made only from oils and fats which are triglycerides and not from any other
kinds of oil (such as engine oil). Chemically, triglyceride consists of three long chain fatty acid
molecules joined by a glycerin molecule. Waste oil is more appealing than using new oil because
refined fats and oils have a free fatty acid (FFA) content of less than 0.1%, in contrary with used and
waste oil, where FFA contents are high. FFAs are formed by cooking, the oil longer and hotter the
oil has been cooked, the more FFAs it will contain. The study reports on biodiesel production from
waste vegetable oil procured from markets where a catalyst (lye) was used to break off the glycerin
molecule and combine each of the three fatty acid chains with a molecule of methanol or ethanol,
creating mono-alkyl esters, or Fatty Acid Methyl Esters (FAME)—biodiesel. In this process of
Transesterification, the glycerin sunk to the bottom and was removed. FFAs interfere with the
Transesterification process inhibiting biodiesel formation. With waste oil more lye had to be used to
neutralize the FFAs.
KEY WORDS: Biodiesel, Waste vegetable oil, Triglyceride, FFAs, Transesterification
Research article
Cite this article:
Chethana G.S, Reddy K Dayakar, Vijayalakshmi (2013), BIODIESEL PRODUCTION FROM
WASTE VEGETABLE OIL, Global J Res. Med. Plants & Indigen. Med., Volume 2(2): 102–109
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 102–109
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Petroleum was formed by geologic
processes dating from the Cretaceous and
Jurassic periods, 90 to 150 million years ago,
when vast amounts of zooplankton, algae, and
other organic material were deposited on ocean
floors. However, the majority of petroleum
now extracted in the range of 85% is used to
produce fuels. Most of these are transportation
fuels such as gasoline, diesel fuel, and jet fuel,
while some, such as fuel oil, liquefied
petroleum gas, and propane, are used for
heating and power generation. Petroleum
accounts for more than 90% of transportation
fuel, but only for 2% of electricity generation.
Increasing worldwide demand for petroleum
will affect the transition in important ways.
Global petroleum demand is currently at 84
million barrels per day, and it is predicted to
increase by 1% to 2% per year, reaching 116
million barrels per day by 2030. Much of this
increasing demand will occur in developing
nations (Howard Frumkin et al., 2009).
Air quality data generated by the Central
Pollution Control Board (CPCB) for 2007
under the National Air Quality Monitoring
Programme (NAMP) presented deadly facts
about air pollution levels in Indian cities.
Centre for Science and Environment has
analysed the official data to assess the state of
air quality and trend in Indian cities. The most
widely monitored pollutants in India are
particulate matter (PM), nitrogen dioxide
(NO2), sulphur dioxide (SO2), and on a limited
scale carbon monoxide. Some of the worst
forms of air pollutions are found in Indian
cities. The Central Pollution Control Board
(CPCB) considers air to be ‘clean’ if the levels
are below 50 per cent of the prescribed
standards for pollutants (Centre for science and
environment, 2012).
Biodiesel is an alternative fuel source made
from renewable resources such as vegetable oil
or animal fat, which is simple to use, gives
clean burning, biodegradable, non toxic, and
essentially free of sulfur and aromatics.
Biodiesel is meant to be used in standard diesel
engines and is thus distinct from the vegetable
and waste oils used to fuel converted diesel
engines. Biodiesel contains no petroleum, but it
can be blended with petroleum diesel to create
a biodiesel blend. It can be used in diesel
engines with no major modifications. Biodiesel
is registered as a fuel and fuel additive with the
U.S.Environmental Protection Agency (EPA)
and meets clean diesel standards established by
California Air Resources Board (ARB). Neat
(100 percent) biodiesel has been designated as
an alternative fuel by the U.S. Department of
Energy (DOE) and the U.S. Department of
Transportation (DOT) (California energy
commission, 2012).Since the passage of the
Energy Policy Act of 2005, biodiesel has been
increasing in the U.S. In Europe, the renewable
Transport Fuel Obligation obliges suppliers to
include 5% renewable fuel in all transport fuel
in the EU by 2010.
This study was undertaken to awaken the
utility of waste Vegetable oil which is a trashed
product from hotels, canteens etc. which after
little chemical treatment can be used as an
efficient bio-diesel. Chemically, triglycerides
contained in vegetable oil or animal fat consists
of three long chain fatty acid molecules joined
by a glycerin molecule. Waste oil is more
appealing than using new oil because refined
fats and oils have a free fatty acid (FFA)
content of less than 0.1%, in contrary with used
and waste oil, where FFA contents are high.
FFAs are formed by cooking, the oil longer and
hotter the oil has been cooked, the more FFAs
it will contain. Hence this study was conducted
to use these waste oils where lye was used to
break the Glycerin chain.
MATERIALS (Keith Addison, 2012)
1 liter of fresh vegetable (sunflower) oil and
waste vegetable oil from a local canteen
was procured.
4.5 g of potassium hydroxide (also known
as lye)
200 ml of ethanol (ethyl alcohol)
10 ml isopropyl alcohol
Glass or plastic container that is marked for
1 liter
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METHODOLOGY
Basic titration
For processing used oil it is essential to
titrate the oil to determine the free fatty acid
(FFA) content and calculate how much
extra lye will be required to neutralize it.
Phenolphthalein indicator was used. 1g of
pure Potassium Hydroxide lye (KOH) was
dissolved in 1 liter of distilled water (0.1%
W/V KOH solution)
In a small beaker, 1 ml of dewatered waste
vegetable oil was dissolved in 10 ml
isopropyl alcohol. The beaker was gently
warmed on a hot water bath; stirred until all
oil dissolved in the alcohol and the mixture
turns clear. 2 drops of phenolphthalein
indicator was added. Using graduated
syringe, 0.1% KOH solution was added
drop by drop to the Oil-alcohol-
phenolphthalein indicator, stirring all the
time, kept stirring. The lye solution was
added until the solution stays pink for 15
seconds. The number milliliters of 0.1% lye
solution used was noted and added to the
3.5 grams of lye (the basic amount of lye
needed for fresh oil). So the total quantity
of lye used to process the Waste vegetable
oil per liter is 4.5 gms (Venkata Ramesh
Mamilla et al., 2011; C.V. Sudhir et al.,
2007).
The production of biodiesel (Keith Addison,
2012)
Fresh Sunflower oil & Waste Vegetable oil
were taken to which the amount of catalyst
to be added was calculated as 4.5 for both.
200 ml ethanol was poured into glass
blender pitcher.
Blender was turned on to its lowest setting
and slowly 4.5 g of potassium hydroxide
(lye) was added. This reaction produced
potassium methoxide.
Ethanol and potassium hydroxide was
mixed until the potassium hydroxide has
completely dissolved (about 2 mins), 1 liter
of waste vegetable oil was added to this
mixture. Similar procedure was followed
for new vegetable oil.
This mixture (on low speed) was blended
continuously for 20 mins to 30 mins.
After completing the procedure the oils
were kept for observation. The bottle of oil
was kept for 2 days, uncovered inside a
rack.
Purification step
Purification of the resultant bio-diesel was
done in accordance with the method explained
by Y. Zhang et al., 2003 & Arjun B. Chhetri et
al., 2008.
Figure 1: The apparatus used for Biodiesel production
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The confirmatory test:
Wash test
150 ml of unwashed biodiesel (settled for
12 h or more, with the glycerin layer
removed) was taken in a half liter glass jar
or PET bottle.
150 ml of water (at room temperature), was
added. Screwed the lid on tight and shaken
it up and down violently for 10 seconds and
was let to settle (figure 2) (Keith Addison,
2012).
Methanol test
25 ml of biodiesel was dissolved in 225 ml
of methanol in a measuring glass. The biodiesel
got dissolved completely in methanol. ―The
biodiesel should be fully soluble in the
methanol, forming a clear bright phase (figure
6) (Jan Warnqvist, 2005).
Figure 2: Wash test for biodiesel
Figure 3: Picturing shows Methanol test carried out for biodiesel, along with biodiesel
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Figure 4: The total amount of Biodiesel obtained
RESULTS
Production results
Biodiesel was obtained after processing the
waste vegetable oil and new sunflower oil.
After 2 days of observation, it was observed
that the biodiesel was on top of the
glycerin, which settled at the bottom. The
amount of biodiesel obtained from waste
vegetable oil was 540 ml from 1 liter and
850 ml of biodiesel from 1 liter sunflower
oil (figure 4).
The purification step
This step was done by washing biodiesel
with water. This was done to remove the
impurities and the incomplete reaction
products like soap.
10 ml of normal tap water was added to
100 ml of biodiesel, shaken vigorously,
allowed for some time and the water was
removed. This was done until we got clear
water indicating that most of the impurities
were removed.
Wash Test
The biodiesel should separate from the
water in half an hour or less, with amber (and
cloudy) biodiesel on top and milky water
below. After a violent 10-second shaking;
biodiesel and water separated cleanly within
minutes. This is quality fuel, a complete
product with minimal contaminants. It was
observed that the clear water was at the bottom
and biodiesel was on the top. This indicates the
positive result for biodiesel for wash test. This
tells that the biodiesel got purified, that is the
oil which underwent incomplete reaction was
removed by wash test. The biodiesel which is
purified stands on the top leaving clear water at
the bottom (figure 5).
Results of Methanol test:
Biodiesel dissolves easily in methanol,
where as vegetable or animal oils and fats
(triglycerides) does not dissolve in methanol.
Any uncovered oil left in the biodiesel will
settle out at the bottom of the tank. 25 ml of
biodiesel was added in 225 ml of methanol. A
clear solution indicates a positive result for
biodiesel (figure 6).
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Figure 5: Positive result for biodiesel by wash Test
Figure 6: Confirmation of biodiesel by Methanol test
DISCUSSION
Biodiesel is an alternative fuel similar to
conventional or fossil diesel. Biodiesel can be
produced from straight vegetable oil, animal
oil/fats, tallow and waste cooking oil. The
process used to convert these oils to biodiesel is
called Transesterification (Ulf Schuchardt et al.
1997).
Biodiesel has many environmentally
beneficial properties. The main benefit of
biodiesel is that it can be described as ‘carbon
neutral’. This means that the fuel produces no
net output of carbon in the form of carbon
dioxide. The (figure 7) below shows the chemical
process for methyl ester biodiesel. The reaction
between the fat or oil and the alcohol is a reversible
reaction and so the alcohol must be added in excess
to drive the reaction towards the right and ensure
complete conversion.
Mixing of alcohol and catalyst
The catalyst used was typically potassium
hydroxide. It was dissolved in alcohol which
acts and enhances the reaction with the oil to
form esters (figure 7) which is nothing but the
crude biodiesel which is in compliance with the
study done by Venkata Ramesh Mamilla et al.,
2011. Excess of catalyst was used to convert
the oil completely into esters. The reaction
happens with vigorous agitation, done using a
mixer. The recommended reaction time was 20
minute to 1 hour. The so formed biodiesel is
kept ideal for 24 to 48 hours under observation.
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Figure no 7: Transesterification of Fatty acids
Separation
Our results on separation step are similar to
the study conducted by Y. Zhang et al., 2003)
which shows the clear separation of the
biodiesel on the top, from the glycerine at the
bottom which is much denser than the
biodiesel. Once its separated from glycerine,
biodiesel is is sometimes purified by washing
with warm water to remove residual catalyst or
soaps, dried and sent to storage, which marks
the end of the process. The process results in
the clear amber-yellow liquid.
CONCLUSION
In the present situation where the natural
resources in the form of fossil fuel are getting
exhausted, it has become very important to
think the alternate source of energy. So
biodiesel is one of the alternate solutions which
are ecofriendly. It is also advantageous over the
pollution caused by petroleum products
because Biodiesel is a biodegradable, non toxic
and virtually free from sulfur and aromatics. A
number of studies have found that biodiesel
biodegrades much more rapidly than
conventional diesel. In this respect, its action is
similar to petroleum diesel fuel. However,
biodiesel does not have the toxicity and the
solvent action that diesel fuel has, so its effects
on animals are expected to be less severe. A lot
of research and development is needed in this
aspect to make the biodiesel easily available to
everyone.
ACKNOWLEDGEMENTS
Authors are thankful to the Head of the
Department, Biotechnology, Dr.Vedamurthy
Anakalbasappa and other lecturers for their
support in completion of this work
successfully.
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Arjun B. Chhetri, K. Chris Watts and M.
Rafiqul Islam (2008), Waste Cooking
Oil as an Alternate Feedstock for
Biodiesel Production., Energies; 1, 3–
18; DOI: 10.3390/en1010003
California energy commission, (2012),
California energy commission retrieved
from
http://www.consumerenergycenter.org/t
ransportation/afvs/biodiesel.html
Centre for science and environment, (2012),
Air pollution, State of Air Pollution in
Indian cities, retrieved from
http://www.cseindia.org/node/207
C.V. Sudhir, N.Y. Sharma and P.Mohanan.,
(2007) Potential of waste cooking oils
as biodiesel feed stock., Emirates
Journal for Engineering Research, 12
(3): 69–75
Gerhard Knothe, Robert O. Dunn and Marvin
O. Bagby, (1997) The Use of Vegetable
Oils and Their Derivatives as
Alternative Diesel Fuels. Oil Chemical
Research, National Center for
Agricultural Utilization Research,
Agricultural Research Service, U.S.
Department of Agriculture, Peoria,
Volume: 666; Chapter, 10; pp-172–208
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 102–109
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Chapter DOI: 10.1021/bk-1997-
0666.ch010
Howard Frumkin, MD, Dr P HJeremy Hess,
MD, MPH Stephen Vindigni., (2009),
Energy and Public Health: The
Challenge of Peak Petroleum., Public
Health Rep; 124(1): 5–19.
Jan Warnqvist, (2005). AGERATEC AB
Biofuel mailing list Re: Quality Test,
retrieved on 12.12.2012 from
http://www.mwil-
archive.com/[email protected]/
msg53363.html
Keith Addison (2012), JOURNEY TO
FOREVER, HONG KONG TO CAPE
TOWN TO OVERLAND, retrieved
from http://www.Journeytoforever.org
Ulf Schuchardt , Ricardo Serchelia, and
Rogério Matheus, (1998). Esterification
of Oils :a review., J Braz, Chem. Soc.
Vol 9, No. 1, 199–200.
Venkata Ramesh Mamilla, M.V.
Mallikarjun,Dr.G Lakshmi Narayana
Rao., (2011), Preparation of Biodiesel
from Karanja Oil., Internal journal of
Mechanical & Production Engineering
Research & Development.Vol 1. (1):
51–69.,
Y. Zhang, M.A. Dubee, D.D. McLean, M.
Kates. (2003) Biodiesel production
from waste cooking oil: 1. Process
design and technological assessment.,
Bioresource Technology 89:1–16.
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
PHYTOCHEMICAL STUDIES ON SMILAX MACROPHYLLA LINN.;
A SOURCE PLANT OF CHOPACHEENI
Jyothi T1*, Acharya Rabinanaryan
2, Shukla C P
3, Harisha CR
4
1Research assistant, ALN Rao Memorial Ayurvedic medical college, Koppa – 577126, Chikkamagalur
District, Karnataka, India 2Associate Professor, Department of Dravyaguna. IPGT& RA, Gujarat Ayurved University, Jamnagar-
361008, Gujarat, INDIA 3Head, Department of Pharmaceutical Chemistry, IPGT& RA, Gujarat Ayurved University, Jamnagar-
361008, Gujarat, INDIA, 4Head, Pharmacognosy Laboratory, IPGT& RA, Gujarat Ayurved University, Jamnagar- 361008, Gujarat,
INDIA
*Corresponding Author: [email protected]
Received: 10/01/2013; Revised: 02/02/2013; Accepted: 05/02/2013
ABSTRACT
Chopacheeni is an important herbal drug widely used in Ayurveda. Chopacheeni is one among in
the red list and in top 20 highly traded medicinal plants in India. Commonly known as Sarsaparilla,
various species are available in the market in the name of Chopacheeni and are rarely Smilax china,
the official source. The plant is considered as a remedy for Syphilis, Rheumatism, Skin diseases and
Gout. Botanically authenticated drug is Smilax china but due to unavailability many adulteration is
coming in the market to avoid this it is attempt to look for substitute of the drug of same genus so
Smilax macrophylla Linn. was used for the phytochemical analysis. TLC of alcoholic extract of drug
on silica gel "G" plate using Toluene (6.5 ml): Ethyl acetate (3.5 ml): Glacial acetic acid (0.2 ml)
showed one spot under 366 nm UV, in 254 nm UV no spots, After spraying with Liebermann
Burchard reagent followed by heating and then was visualized in day light which showed 2
prominent spots are seen.TLC using Chloroform (9.5 ml): Methanol (0.5 ml) showed two spots
under 366 nm UV ,in 254 nm UV no spots were seen. After spraying, it showed one prominent spot.
In HPTLC chromatogram showed 2 prominent spots in short wave UV 254 nm, one prominent spot
in long wave UV 356 nm and 3 prominent spots at 400 nm.
KEYWORDS: Smilax macrophylla, Chopacheeni, Madhusnuhi, Phytochemical Analysis
Research article
Cite this article:
Jyothi T, Acharya Rabinanaryan, Shukla C P, Harisha CR (2013), PHYTOCHEMICAL STUDIES
ON SMILAX MACROPHYLLA LINN.; A SOURCE PLANT OF CHOPACHEENI, Global J Res.
Med. Plants & Indigen. Med., Volume 2(2): 110–117
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Smilax china Linn. of the Family
Smilacaceae) is an important herbal drug
widely used in Ayurveda. It is used for the
treatment of Phiranga roga, Upadamsha,
Vatavyadhi, Vrana (Vaidya Bapalal, 2005). It is
said that the drug is similar to Ashwagandha
(Sharma PV, 2005) in its properties and action.
It is one among the red listed plants and in top
20 highly traded medicinal plants in India
(http://www.megforest.gov.in). Now a day, due
to commercialization and other economic
interests, the pharmaceutical industry uses
various source plants for the drug Chopacheeni.
The availability of genuine samples is a
burning issue since it is enlisted as endangered
species (www.iucnredlist.org) and demand
surpasses production causing confusion in the
end users about the quality and safety also
therapeutic ambiguity. Various species of
Smilax are used as source plant for
Chopacheeni; S. China Linn. , S. macrophylla
Linn., S. zeylanica Linn. , S. regelli to name a
few. S. china is formerly considered as the
authentic identification as per The Wealth of
India (Anonymous, 1950). Due to non
availability and possible adulteration of cheaper
substitutes, the therapeutic efficacy is obscure.
Hence there is an urgent need to explore the
raw drugs for their quality through
phytochemical investigations to establish the
authenticity and logical reasoning behind
multiple source plants of Chopacheeni. In the
current research, roots and rhizomes of Smilax
macrophylla Linn. which is a potential source
for Chopacheeni was evaluated phyto-
chemically for available active ingredients and
their strength. Pharmacognostical investigation
with macerate and powder study details and
HPTLC finger printing of the rhizome and
roots of Smilax macrophylla Linn. which helps
in identification of crude drug is not available.
Hence the present study has been carried out
with following Aims & Objectives;
Pharmacognostical and Phytochemical analysis
of Smilax macrophylla Linn.
The vernacular names (Prajapati ND et al.,
2003) of Chopacheeni (Smilax china) are:
Sanskrit- Chopacheeni, Dvipantharavacha,
Madhusnuhi (Chopra RN et al., 1956); English-
Sarsaparilla, China root; Hindi- Jangli,
Austibab, Chopachini; Marathi- Guti; Tamil-
Malaith, Tamarai, Parangichekkarda; Telugu-
Kondata mora, Malkaltamora; Kannada-
Kaduhambu; Gujarati- Chopachini; Assamese-
Aslussini; Bengali- Topachini
Smilax macrophylla Linn. is a large more or
less prickly climber (Haines HH, 2000)
growing in Himalaya eastwards from Kumaon
at Assam, Bengal, Burma & South to Central
Konkan extending to Java. The Stem is
Smooth, striate (lines or several angled), armed
with a few small distant prickles or almost
unarmed; roots are rope-like originate from a
short rhizome (Haines HH, 2000).
MATERIALS & METHODS
Plant Material:
Fresh roots and rhizomes were collected
from the forests of Odisha during the month of
November 2009. Botanical identification was
done by expert taxonomists using local floras
(Haines HH, 2000) and found to be Smilax
macrophylla Linn. Voucher Ref no. IPGT&RA
- 301). The collected samples were washed
with potable water and chopped in to small
pieces which were dried in shade, powdered
and used for scientific evaluation.
Pharmacognostical Study (Kokate CK, et
al., 2008, Anonymous 2000, Trease and
Evans, 2009): Morphological,
Organoleptic, Microscopic and
Histochemical study of the powdered drug
was done as per the guidelines of
Ayurvedic pharmacopoeia of India.
Phytochemical study (Kasture AV et al.,
2009, Anonymous 2000, Stahl E. 2005):
Smilax macrophylla Linn. were analyzed by
Physicochemical, Qualitative, Quantitative
parameters. Chromatographic fingerprinting
and Ultra-Violet Spectroscopy were carried
out. TLC is mentioned as a primary tool for
identification as part of monographs on all
medicinal plants. Methanolic extract was
used for the spotting of the TLC plate
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
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(Silica gel G Pre-coated plates). The solvent
systems used in the study were Chloroform:
Methanol and Toluene: Ethyl acetate: acetic
acid. The sample extract was made to run
on silica plate in various ratios. The ratio of
9.5:0.5 and 6.5:3.5:0.2 respectively has
given good separation on trial method.
Hence these systems are adopted for
Chromatographic evaluation of the sample.
Acetic acid was added for the second
system for better separation. The resulting
TLC pattern was viewed under long wave
UV light at 366 nm and Short wave at 254
nm. Then after spraying with the
Liebermann Burchard reagent and drying in
a hot air oven and the number of spots
viewed under daylight (Table no.4).
Picture No. 1 Pharmacognostical Study of Smilax macrophylla Linn
Picture No. 2 Pharmacognostical Study of Smilax macrophylla Linn.
A.Rhizome, B. Root, C. Cork, D. Acicular Crystals, E. Scalariform Vessels, F. Scleroids,
G. Tannins & Starch Grains, H. Fiber, I. Parenchyma, J. Tracheids, K. Pitted Vessels,
L. Reticulate Vessel
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
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RESULTS & DISCUSSION:
Pharmacognostical Study: (Picture No.2)
Morphological Study: Rhizomes were 5–6
× 3–4 cm in size, Root 20–23 × 1.5–2 cm in
length Cylindrical & tapering towards apex,
externally brownish and internally pinkish
in color with rough and woody surface, and
fracture coarsely fibrous.
Organoleptic characters: The powder was
reddish brown in color, with characteristic
odor, slight bitter in taste and fibrous in
texture.
Powder Microscopy: The dried powder
was mounted with distilled water to detect
the Starch Grains, Cork, Simple Fiber,
Acicular crystals, Scalariform vessels,
Scleroids, Reticulate Vessels, Pitted
Vessels, Parenchyma, Tracheids and
Tannin. When stained with Iodine solution,
Dil. FeCl, Conc. HCl and Phloroglucinol
with Conc. Hcl, Showed Starch grains
(Blue), Tannins (Bluish black), Crystals
(Effervescence) and Lignified cells (Pink)
respectively.
Physicochemical Parameters: The sample
was evaluated for physicochemical
parameters like Loss on drying, Total Ash
value, Acid insoluble ash, water, alcohol,
chloroform, acetone soluble extractive
values and for pH value (Table No. 1). The
percentage of moisture content was
9.40%w/w, total ash 2.45%w/w, acid
insoluble ash 0.15%w/w; Water soluble
extractives 31.25%w/w, Alcohol soluble
extractives 19.30%w/w, Chloroform
soluble extractives 0.1%w/w and Acetone
soluble extractives 8.58%w/w. pH was
5.28. Low total ash and Acid insoluble ash
signifies low levels of inorganic matter and
silica content. The high solubility of the
sample in water denotes that drug is best
suited for extraction with water or water
based preparations. The negligible presence
of Volatile oils is also in favor of thermal
extractions with water.
Qualitative chemical tests: Qualitative
chemical tests for different functional units
were estimated using water, methanol,
chloroform and acetone soluble extractives
of Smilax macrophylla Linn.
Carbohydrates, Reducing sugars, proteins,
Amino acids, saponins, alkaloids,
Flavonoids and Tannin were qualitatively
investigated. All functional units were
present in water soluble extractive except
amino acids and Flavonoids as these are the
basic functional units necessary for
metabolism in herbs. (Table No.2)
Table No: 1 Physico Chemical Parameters
Sr.
No.
Parameter Smilax macrophylla
Linn.
1 Loss on Drying 9.40%w/w
2 Total Ash 2.45%w/w
3 Acid Insoluble Ash 0.15%w/w
4 Water Soluble Extractives 31.25w/w
5 Alcohol Soluble Extractives 19.30%w/w
6 pH 5.28
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Table No. 2 Qualitative chemical tests of Smilax macrophylla Linn. for different functional
units in various solvent systems
Sr.
No.
Test Water Methanol Chloroform Acetone
1 Carbohydrates-Molish’s
test
Positive Positive Positive Positive
2 Reducing Sugars-Fehling’s
test
Strongly Positive Strongly
Positive
Positive Positive
3 Proteins-Biuret test Positive Negative Negative Negative
4 Amino acid- Ninhydrin test Negative Negative Negative Negative
5 Saponins-
Foam test
Moderately
Positive
Positive Negative Negative
6 Flavonoids-Shinoda test Negative Positive Negative Negative
7 Alkaloids- Wagner’s test Positive Positive Negative Negative
8 Tannins- FeCl3 Strongly Positive Positive Negative Positive
Table No.3 Quantitative estimation
Sr. No. Parameter Smilax macrophylla Linn.
1. Total Volatile oils Trace
2 Total Alkaloids 0.08%w/w
3 Total Tannins 8.28%w/w
4 Total Saponins 22.85%w/w
Quantitative estimation: Traces of total
Volatile oils, 0.08%w/w of total Alkaloids,
8.28%w/w of total Tannins and 22.85%w/w
total Saponins were observed in the sample.
Saponins are high molecular weight
glycosides, consisting of a sugar moiety
linked to a triterpene, steroid or steroid
alkaloidal aglycone (Natural Remedies).
Aglycone portion of the saponin is called as
sapogenin. Triterpene saponins are the most
common type in the plant kingdom. They
show hemolytic activity and have bitter
taste. Majority of the pharmacological and
clinical action may be linked to these
saponins in the case of Chopacheeni.
(Table No.3).
Thin Layer Chromatography Study:
Thin layer chromatography of Methanol
Extract of Smilax macrophylla Linn.
Powder of the sample weighing 5 g were
taken with 100 ml of alcohol and kept for
twenty-four hours. Filtrate was prepared
and evaporated till it was dried in a flat-
bottomed shallow dish and concentrated on
water bath to volume of requirement. TLC
of alcoholic extract of drug on silica gel
"G" plate using Toluene (6.5 ml): Ethyl
acetate (3.5 ml): Glacial acetic acid (0.2 ml)
showed one spot under 366 nm UV at Rf
0.23. Where as in 254 nm UV no spots
were seen. After spraying with Liebermann
Burchard reagent followed by heating and
then was visualized in day light which
showed 2 prominent spots at Rf 0.70 and
0.82. TLC using Chloroform (9.5 ml):
Methanol (0.5 ml) showed two spots under
366 nm UV at Rf 0.04 and 0.83. Where as
in 254 nm UV no spots were seen. After
spraying, it showed one prominent spot at
Rf 0.83.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
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Table No. 4 Thin Layer Chromatography of Smilax macrophylla Linn.
Mobile Phase Detection condition No.of
Spots
Rf Value
Chloroform:
Methanol (9.5:0.5)
254 nm UV 0 -
366 nm UV 2 0.04, 0.83
After spray with Liebermann
Burchard reagent
1 0.83
Toluene: Ethyl
acetate: acetic acid
(6.5:3.5:0.2)
254 nm UV 0 -
366 nm UV 1 0.23
After spray with Liebermann
Burchard reagent
2 0.70,0.82
Table No. 5 High Performance Thin Layer Chromatography of Smilax macrophylla Linn.
Mobile Phase Detection condition No.of
Spots
Rf Value
Chloroform:
Methanol (9.5:0.5)
254 nm UV 2 0.08, 0.56
366 nm UV 1 0.08
400 nm UV 3 0.10, 0.28, 0.70
Table No.6 UV Spectrophotometry of Smilax macrophylla Linn.
Sample Peak Wavelength
in nm
Absorbance
Smilax macrophylla Linn. 1 236 3.161
1 275.6 3.464
Picture No. 3 HPTLC of Smilax macrophylla Linn.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Picture No. 4 UV Spectrophotometry of Smilax macrophylla Linn
High-Performance Thin Layer
Chromatography Study: Methanol
extract of Smilax macrophylla Linn. was
spotted on pre-coated silica gel GF 60254
aluminium plate as 5 mm bands, 5 mm
apart and 1 cm from the edge of the plates,
by means of a Camag Linomate V sample
applicator fitted with a 100 μL Hamilton
syringe. Chloroform (9.5 ml) and Methanol
(0.5 ml) (v/v) (20 ml). The Chamber was
saturated for 45 min, Development Time
taken was 20 min and the development
distance was 4.8cm. After development,
Densitometric scanning was performed
with a Camag T.L.C. scanner III in
reflectance absorbance mode at 254 nm,
366 nm and 400 nm under control of win
CATS software (V 1.2.1 Camag) (Picture
No.2). The slit dimensions were 6 mm x
0.45 mm and the scanning speed was 20
mm s-1
.
However, chromatogram showed 2
prominent spots at Rf 0.08 and 0.56 in short
wave UV 254 nm, one prominent spot at Rf
0.08 in long wave UV 356 nm and 3
prominent spots at 0.10, 0.28, 0.70 at 400
nm. (Table No.5 and Picture No. 3)
UV Spectrophotometry: The spectrum was
measured by placing the sample solution
into the Shimadzu UV-160 Double beam
spectrophotometer. Based on the UV
Spectrophotometric analysis, the peaks,
wavelengths and absorbance are shown in
Table No.6. (Picture No.4)
CONCLUSION
Presence of more acicular crystals,
Scalariform vessels, Scleroids, Reticulate
Vessels, Pitted Vessels, is the identified
character of Smilax macrophylla. The
preliminary phytochemical analysis of the
rhizome and root of Smilax macrophylla
revealed the presence of Carbohydrates,
Reducing sugar, saponin, protein, alkaloids and
Tannins. The sample has got highest solubility
in water followed by methanol. Hence drug is
best suited for extraction with water or water
based preparations. The Chromatographic
finger printing was developed which could be
useful for researchers to carry out further
researches. The study is expected to be useful
for quality control of sample and also will be a
useful guide in deciding the source for
Chopacheeni looking in to lack of availability
and endangered status of the official source
plant Smilax china Linn.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 110–117
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
REFERENCES
Anonymous. (2000), Protocol for testing of
Ayurveda, Siddha & Unani medicines.
Ghaziabad: Pharmacopoeial laboratory
for Indian medicines, Department of
AYUSH, ministry of health & family
welfare, Government of India.
Anonymous. (1950), The wealth of India, Vol.
VIII. New Delhi: Council of Scientific
and Industrial Research;. pp 365.
Chopra RN, Nayar SL, Chopra IC, (1956),
Glossary of Indian Medicinal Plants.
New Delhi: Council of Scientific and
Industrial Research; pp 228.
Haines HH. (2000), Botany of Bihar and
Orissa. Part V-VI. Dehradun: Bishan
singh Mahendrapal Singh; pp 1085.
http://www.megforest.gov.in/activity_medic_pl
ants.htm accessed on 3/23/2011
IUCN (2009), IUCN Red List of Threatened
Species. Version 2009.2.
<www.iucnredlist.org>. Downloaded
on 01/29/2010
Kasture AV, Mahadik Kr, More HN, Wadodkar
SG. (2009), Pharmaceutical Analysis.
18th Ed. Vol.II, Pune: Nirali Prakashan
Kokate CK, Purohit AP, Gokhale SB. (2008),
Chapter 6. Pharmacognosy. 42nd
Ed.
Pune: Nirali Prakashan; pp 6.1–6.45.
Natural Remedies, Master Document on
Tribulus terrestris, Quality control
department, Natural Remedies,
Bangalore
Prajapati ND, Purohit SS, Sharma AK, Kumar
T, (2003), A Handbook of Medicinal
Plants – A Complete source book, 1st
Ed. Jodhpur: Agrobios; pp 477.
Sharma PV, (2005), Dravyaguna Vijnana, Vol.
2, Varanasi, Chaukhambha Bharatiya
Academy; 804
Stahl E. (2005), Thin-Layer Chromatography.
2nd Ed. Berlin: Springer;
Taylor, L., The Healing Power of Rainforest
Herbs downloaded from
http://www.rain-tree.com/book2.htm.
On 10/29/2010.
Trease and Evans. (2009), Pharmacognosy.
16th Ed. New York: Elsevier Saunders;
pp 309–10.
Vaidya Bapalal (2005), Nighantu Adarsha,
Varanasi, Chaukhambha Orientalia,Vol.
II: 643.
Source of Support: Nil Conflict of Interest: None Declared
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
SELECTION OF MEDICINAL PLANTS FOR THE MANAGEMENT OF
DIABETIC FOOT ULCER; AN AYURVEDIC APPROACH
Pampattiwar S P1*, Adwani N V
2, Bulusu Sitaram
3, Paramkusa Rao M
4.
1, 2 P.G. Scholar – final Year, P.G. Dept. of Dravyaguna, T.T.D‟S S.V. Ayurveda College, Tirupati, Andhra
Pradesh, India 3, 4
Professor (UG), Dept. of Dravyaguna, T.T.D‟S S.V. Ayurveda College, Tirupati, Andhra Pradesh, India
*Corresponding Author: E mail: [email protected]; Mobile: +91 9700307493
Received: 05/01/2013; Revised: 08/02/2013; Accepted: 10/02/2013
ABSTRACT
Diabetic foot ulcer is one of the major complications of Diabetes mellitus. It can lead to
amputation of leg also. Diabetes mellitus is one such metabolic disorder that impedes normal wound
healing because of altered protein and lipid metabolism and abnormal granular tissue. This literary
review was done to provide an effective management in cases of non healing ulcer. It is proposed
that for the treatment of such patients common herbs explained in “Prameha Chikitsa”, “Kushtha
Chikitsa” and “Vrana Chikitsa” can be useful. This is a new approach by which one can select the
herbs for the treatment of diabetic foot ulcer. Due to this approach new formulations can be
formulated for diabetic foot ulcer which can be beneficial to them. A list of drugs mentioned in
treatment of above diseases is prepared from Ashtanga Hridaya and their activity checked with
ongoing clinical research.
KEYWORDS: Diabetic foot ulcer, Prameha, Kushtha, Vrana
ABBREVIATIONS: A.H. SU- Ashtanga Hridaya Sutrasthana; WHO – World Health Organization
Review article
Cite this article:
Pampattiwar S P, Adwani N V, Bulusu Sitaram, Paramkusa Rao M (2013), SELECTION OF
MEDICINAL PLANTS FOR THE MANAGEMENT OF DIABETIC FOOT ULCER; AN
AYURVEDIC APPROACH, Global J Res. Med. Plants & Indigen. Med., Volume 2(2): 118–125
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
INTRODUCTION
Diabetic foot is the most common
complication of diabetes greater than
retinopathy, neuropathy, heart attack and stroke
combined. (Marvin E Levin, 1994) According
to WHO, the foot of Diabetic patient has
potential risk of pathological consequences
including infection, ulceration and or
dysfunction of deep tissues associated with
neurological abnormalities, various degrees of
peripheral vascular disease and metabolic
complications of diabetes in lower limb.
(Robert G Frykberg, 2000)
The Diabetic foot is essentially vulnerable
to amputation because of frequent
complications of peripheral neuropathy (PN),
infection and peripheral arterial disease (PAD).
(Marvin E Levin, 1994) A combination of this
triad leads to gangrene and amputation. Most
of these are result of PN (peripheral
neuropathy) and insensate foot which leads to
painless trauma and ulceration.
The relation between Diabetic neuropathy,
the insensitive foot and foot ulceration was
recognized by Pryce, a century ago. He stated
that, “It was abundantly evident that the actual
cause of perforating ulcer was peripheral nerve
degeneration and that diabetes itself played
active part in causation of perforating ulcer”.
(Pryce TD, 1887)
For instance wound infection has been one
of the major impediments in the process of
wound healing and after invention of
antibiotics; it was thought that this problem
would be conquered. Since then several
antibiotics in form of systemic and local use
have been tried but problems of wound healing
remains as such. Apart from this, antibiotics
have their adverse side effects.
There are list of complications occurring as
a result of taking hypoglycemic drugs like chest
pain, irregular heartbeat, difficulty in breathing
and erectile dysfunction. (Seppo Lehto, 1996).
To avoid above complications, it would be
better to go with herbal drugs for the
management of Diabetic foot. Keeping in view
of aforesaid problems, ancient literature was
explored to throw light regarding the wound
and its management with the help of medicinal
plants.
MATERIALS
Though the direct description of Diabetic
foot ulcer is not available in Samhitas but it is
found in „Vrana Paratishedha Adhyaya‟ of
AstangaHridaya (Tripathi, 2007). Here
Vagbhata denoted that if the patient of Vrana
(ulceration) is suffering from Kushtha (skin
disease) or malnutrition or poisoning or
Prameha (diabetes) then that Vrana (ulcer) is
difficult to treat. This quotation clearly verdicts
the reference of Diabetic foot ulcer has been
manifested and said to be almost incurable.
Pathogenesis sequence of diabetic foot ulcer
Classically all the Hetus (causative factors)
described for Prameha are responsible for
vitiation of Kapha, Mutra, Meda. So, Prameha
is vitiated Kapha predominant disease
(Tripathi, 2007). Vitiated Kapha vitiates Kleda
(moisture), Sweda (sweat), Medo-dhatu (fat),
Rasa Dhatu (plasma) and Mamsa Dhatu
(muscle) in body. As vitiated kapha vitiates
Mamsa Dhatu (muscle) it loses its „SAARATA‟
(consistency). This thing creates problem in
healing of Diabetic foot ulcer.
„Twachi Swapa‟ (numbness) and
„Vrananam Shighrotpatti Chirastithi‟
(immediate onset of ulcers and become
chronic), these two symptoms are present in
„Kushtha nidana‟ (etiological factors of Skin
diseases) as Purvarupa (Premonitory
symptoms) of Kushta (Tripathi, 2007). These
two can be correlated with insensate foot and
rapid spreading with fixed nature symbolising
necrotising fascitis involving deeper related
tissues. Hence, vitiation of Rakta Dhatu (blood)
occurs in Diabetic foot. All these references
denote various complications of Diabetic foot
ulcer with bad prognosis nature. There are
some common factors found in „Prameha‟
(Diabetes), „Kushta‟ (skin disease) and „Vrana‟
(ulcer) with respect to Hetus (causative
factors), Doshas (humors) and Dushyas
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
(Vitiated tissue). The common causative factors
are intake of excess curd, new cereals, jaggary,
meat and milk. The common vitiated dosha
(humors) in Prameha and Kushta is mostly
Kapha and common vitiated tissues are Rakta
(blood), Mamsa (muscles) and Lasika (lymph).
From above description, it is clear that there
is much similarity present between Prameha,
Kushtha, and Dushta vrana. Therefore for the
treatment of Diabetic foot ulcer common herbs
described in treatment of foresaid diseases
should be selected. Vitiation of Kleda
(moisture) is also an important factor in the
Samprapti (pathology) of Diabetic foot.
Selection of herbal drugs for the treatment of
Diabetic foot ulcer depends on following
treatment principles like Vrana shodhana
(purifying the wound), Vrana ropana (wound
healing purpose) and blood purifier. (Tripathi,
2007)
Common herbal drugs used in the treatment
of Prameha, Kushtha and Vrana
In the treatment of Prameha pidika, Eladi
gana is used for Vranaropana (wound healing
purpose), Aragwadhadi gana is used for
Udwartrana (rubbing herbal powder against
body), Asanadi gana is used for Parisheka
(pouring) and Vatsakadi gana is used for
internal administration.
(Tripathi, 2007).
Surasadi and Aaragwadhadi ganas are
indicated for Kshalana purpose (Tripathi,
2007). These ganas (groups) can hold an
important place in the treatment of diabetic foot
ulcer. All the Ganas explained above are
commonly indicated in Prameha, Kushtha and
Dushta vrana.
TABLE NO. 1 The properties of these ganas are as follows:
Name of Gana Indications Action
Eladi Gana
(Tripathi,2007)
(A.H. SU 15/43-44)
Vrana prasadana,
Kandupidikakota nashana
It purifies blood.
It has good healing
property.
Aragwadhadi Gana
(Tripathi,2007)
(A.H. SU 15/17-18)
Prameha ch dushta
vranashodhana…
It should be used
externally.
But, can be used orally
also.
Asanadi Gana
(Tripathi,2007)
(A.H.SU. 15/19-20)
Shwitrakushthakapha…krimin…
Prameha ch medo dosha
nibarhana…
It is indicated orally.
But, in “prameha pidika
chikitsa” it is used for
parisheka.
Vatsakadi Gana
(Tripathi,2007)
(A.H. SU. 15/33-34)
kapha meda
Its action is on rasa
dhatu, and medo dhatu.
Therefore it is indicated
orally.
Surasadi Gana
(Tripathi,2007)
(A.H. SU 15/30-31)
shleshma
medakriminishudana…
vranashodhana
Anti-microbial.
It can be used externally.
Arkadi Gana
(Tripathi,2007)
(A.H. SU 15/28-29)
krimikushta visheshat
vranashodhana
Anti-microbial.
It can be used externally.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
TABLE NO. 2 The properties of these ganas are as follows:
The above (Table no. 1) Ganas (group of
drugs) explained can be used in initial stages of
Diabetic foot when secretions are present. For
deeply penetrated Diabetic foot ulcers,
Nyagrodhradi and Padmakadi Ganas (Table
no. 2) are indicated.
Healing of ulcer (Vrana Ropana),
secretions have ceased and infections stage is
over can be done with the use of these Ganas.
These ganas will help in the regeneration of
healthy tissue. Most of the herbs explained in
above Ganas are not available or controversial
like Madhurasa, Kadar etc. Therefore, good
result may hardly be achieved using one gana
in treatment. Hence, to overcome the above
drawback new method can be implemented.
For that instead of using whole gana with the
help of common herbs present in above
explained ganas (Eladi, Aragwadhadi, Asanadi
Gana, Vatsakadi Gana, Surasadi Gana, Arkadi,
Nyugrodhradi Gana, Padmakadi Gana) may be
used. (Table no. 3).
TABLE NO. 3 common herbs present in above explained ganas and their properties:
Nyagrodhradi Gana
(Tripathi,2007)
(A.H.SU 15/41-42)
vranya…
Bhagna sadana…
Most of the herbs present in this
gana are astringent in taste.
Therefore they are Rakta
shodhaka (Blood purifier)
Kledashoshaka (absorbs
secretions) and „Vranaropaka‟
(wound healers)
Padmakadi Gana
(Tripathi,2007)
(A.H.SU 15/12)
brihmana… Herbs found in this gana are
Vata-pittashamaka and
Raktaprasadaka (Blood
purifiers)
Name / Latin name Ayurvedic literature Prayojyanga with active
principle
Current Researches
with references
1) Patha
Cissampelos pareira
Linn.
(From Aragwadhadi,
vatsakadi)
Bhagnasandhankrit..
(Vaidya Bapalal,2007)
Kushthanu…(VaidyaBa
palal, 2007)
Kriminut(Chunekar,201
0)
Sandhaniya
(Sastri,2008)
Vranaan..(Chunekar,
2010)
Moola (root) – pelosine or
Berbeerine 0.5%
Blood purifying (Khare)
Anti-inflammatory, anti-microbial
(N. Savithramma et al.,2011)
2) Karanja
Pongamia glabra
Vent.
(From Aaragwadhadi,
Arkadi)
Vranam
hanta…(Vaidya
Bapalal,2007)
Krimim hanta…(Vaidya
Bapalal,2007)
Kushtam… (Vaidya
Bapalal,2007)
Moolatwak –(rootbark)
Karajin/ Demethoxy karanjin
(Sangwan et al., 2010)
Puspa(flower) – Pongamin/
Quercitin
Anti-microbial, Anti-inflammatory,
Anti-hyperglycemic, Anti lipid
peroxidase, decreases in level of
blood sugar. Increase in glucose 6-
phosphatase activity and enhancing
anti-oxidant status Flowers are used
in diabetes. (Khare)
3) Chirabilva
Holoptelia integrifolia
Planch.
(From Aragwadhadi,
Arakadi, Asandi)
Kushta twak dosha
vrana
nashana….(Vaidya
Bapalal,2007)
Seed powder dissolved in water
showed hypoglycemic activity in
alloxinised rabbits. (Khare,)
Anti-microbial, anti-oxidant, anti-
inflammatory properties of leaves
(Sharma et al., 2009)
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
DISCUSSION
Diabetic foot ulcer is a common
complication in Diabetic patients which is
prevailing and disturbing the individual‟s
routine and certainly lowering quality of life.
To avoid complications like gangrene and
amputation, there is a need to develop a new
treatment protocol which is simple and cost
effective.
4) Kramuka
Areca catechu Linn.
(From Surasadi,
Asanadi)
kledamalapaha…(Sastri
, 2005)
Catechin, Arecoline (0.17%),
Arecaidine (Patil et al.,2009)
Stimulation of nervous system,
anti-oxidant activity, anti-microbial
activity, hypoglycemic activity.
(Patil et al.,2009)
5) Vidanga
Embelia ribes Burm.f.
(From Surasadi,
Vatsakadi)
Krimighna,
Embeline
(2-3%), Christembine (K.
Haq et al.,2002)
Wound healing activity, anti-
Diabetic activity , seeds-Blood
purifying (M. Chitra, 1980)
6) Murva
Marsdenia
tenacissima W.& A.
(from Aragwadhadi,
Vatsakadi)
Mehanut
(Chunekar,2010)
Kushthapaha.(Chuneka
r,2010)
API recommends bark in lipid
disorders, Anti-hypoglycemic
activity
-glucosidase inhibitor
(Bacchawat, 2011)
7) Arjuna
Terminalia arjuna
(From Nyagrodhradi,
asanadi)
Medomehavranam
hanti (Chunekar, 2010)
Twak (stem bark) –
Arjuentine, Frideline
Glycoside, β-cystocetrol
(Chander Ramesh et al.,
2004)
-glucosidase inhibitor
(Bacchawat 2011)
Anti-inflammatory and immuno
modulator. (Halder et al., 2009)
8) Palasa
Butea frondosa
Koen.ex Roxb.
(Asanadi,
Nyagrodhadi)
Kushthanut… (Shastri,
2005)
Pramehanut. (Shastri,
2005)
Beeja (seed) – Palasonin
Twak (stem bark) –
Kinotannic acid (Borkar,
2010)
Fruit: Hypoglycemic and Hypo
lipidemic activity
Anti-oxidant (Miriyala et al., 2008)
9) Indrajava
Holarrhena
antidysenterica Wall.
(From Argwadhadi,
Asanadi)
seeds – Conessin, Kurchicine
(Lather, 2008)
Seeds: Anti-Diabetic activity,
reduces LDL, VLDL, Elevation of
glucose-6-phosphatase activity,
reduces blood sugar level help in
stimulation of -cells of islets of
langerhans.
10) Bharangi
Clerodendron
serratum Spreng.
(From Arkadi,
Surasadi, Vatsakadi)
Vranakrimighni…
(Vaidya Bapalal, 2007)
Moola twak (stem bark) –
Phenolic glycoside, saponine
(Kajaria et al., 2011)
Antimicrobial, antioxidant,
antiDiabetic (Shrivastava et al.,.
2007)
11) Agaru
Aquillaria agallocha
Roxb.
(From Asanadi,
Eladi)
Kushthanut… (Vaidya
Bapalal, 2007)
Heartwood-Sesquiterpene
alcohol (Bhuiyan et al.,
2009)
12) Ela
Elettaria
cardamomum Maton.
(From Vatsakadi,
Eladi)
Oil – Cineol, terpineol,
terpinene, Limonene,
Sabinene (Atta et al., 2000)
Antioxidant
13) Rohisha trina
Cyambopogon
martini(Roxb) Wats.
(From Eladi,
Surasadi)
Oil - Geraniol
(80-94%), (Ginger oil)
(Dubey et al.,2003)
Antiseptic, wound healer.
Global J Res. Med. Plants & Indigen. Med. | Volume 2, Issue 2 | February 2013 | 118–125
Global Journal of Research on Medicinal Plants & Indigenous Medicine || GJRMI ||
Diabetic foot ulcer is not directly explained
in Samhitas (Classical lexicons). Its Samprapti
lies between Prameha, Kushtha and Dushta
Vrana. Common herbs explained in the
treatment of the above mentioned diseases
would be better to use for the treatment of
Diabetic foot ulcer. These common herbs are
Patha (Cissampelos pareira Linn.), Karanja
(Pongamia glabra Vent.), Chirabilva
(Holoptelia integrifolia Planch.), Kramuka
(Araca catechu Linn.), Vidanga (Embelia ribes
Burm.f.), Murva (Marsdenia tenacissima W.&
A), Arjuna (Terminalia arjuna), Palasa (Butea
frondosa Koen.ex Roxb.), Indrajava
(Holarrhena antidysenterica Wall.), Bharangi
(Clerodendron serratum Spreng), Agaru
(Aquillaria agallocha Roxb.) and Rohisha
Trina (Cyambopogon martini(Roxb) Wats).
These herbs can be used as a single drug or in
combination in the treatment of Diabetic foot
ulcer. These can be used externally
(Shrivastava et al., 2011) orally or both.
CONCLUSION
Management of Diabetic foot should be
multipronged attack like controlling blood
sugar level, preventing infection and avoiding
peripheral nerve tissue damage is crucial.
Hence in this study, an attempt has been made
with available herbal drugs have been proved
since time memo rid thoroughly and classified
symptomatically keeping complications of
Diabetic foot ulcer in mind. Regarding the
treatment of an ulcer, two steps in Ayurveda
are very important which are the Shodhana and
Ropana and they have similar concept with
debridement, dressing and elevation of wound
as mentioned in modern medicine. Common
herbs explained in the treatment of Prameha,
Kushtha and Dushta Vrana were reviewed
because they have potent medicinal property,
less or negligible adverse effect. This study or
various aspects of diabetic foot ulcer with
single drug regimen specifically on various
intricacies of disease particularly could prove
beneficial to mankind.
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Source of Support: Nil Conflict of Interest: None Declared
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