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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

mailto:[email protected]



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



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).



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.



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.



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




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]


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




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



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



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).



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.



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-Garcıá 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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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]


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




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).



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.



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



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.



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).



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;



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



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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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]


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




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



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



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



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).



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.



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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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]


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




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



(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



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



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.



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.



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.



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

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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;

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Varanasi, Chaukhambha Orientalia,Vol.

II: 643.





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


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



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



(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.



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)



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.



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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