Journal of Al-Nahrain University Vol.15 (3), September, 2012, pp.7-14 Science
Identification of Dandelion Taraxacum officinale Leaves Components and Study
Its Extracts Effect on Different Microorganisms
Abdul Kadir M. N. Jassim*, Safanah Ahmed Farhan** and Omar Mohammed Noori***
*
Department of Chemistry, College of Science, Al-Mustansiriyah University, Baghdad-Iraq.
**
Polymer research center, College of Science, Al-Mustansiriyah University, Baghdad-Iraq.
***
Industries of Chemical and Physical Materials, Ministry of Science and Technology.
Abstract
Natural plant Dandelion Taraxacum officinale have been used as a phytomedicine, in this study,
the chemical components of the Dandelion Taraxacum officinale leaves in watery and alcoholic
extracts were identified. The results showed that watery extract was alkaline (presence of alkaloids)
while the alcoholic extract was acidic in these contained: glycosides, alkaloids, phenolic
compounds, tannins, flavonoids and proteins, while the saponins and resins were not found.
The result also showed that high concentrations of the following trace elements were found in
the leaves (K ,Ca , Na ,Fe) with 185.1, 22, 19.5, 11.2 ppm, respectively and low concentrations of
(Zn, Cd, Cu) with 6.3, 1.3, 0.2 ppm, respectively. The effect of these extracts on the different
microorganisms were studied. It has been found that the concentration 0.5 mg/ml was effective on
the inhibition of the growth of the intended bacteria (for both extracts) especially Gram positive,
Staphylococcus aureus and the alcoholic extracts with concentration 0.5 mg/ml was more effective
on the Gram negative, E.coli, than the watery extract, while less than 0.1 mg/ml failed to inhibit any
microorganisms. The High Performance Liquid Chromatography (HPLC) was used to identify
some flavonoids as compared to standard one; the analysis showed that both kaempferol and morin
were absent.
Keyword: Dandelion Taraxacum officinale, Plant extraction, Trace elements, HPLC.
Introduction at the same time as a cleanser of the blood and
Herbs have been used for a large range of liver. An active substance of dandelion
purposes .They are found to be potent sources reduces serum cholesterol and triglycerides
of natural components .Some have been used because it intensifies bile secretion [1,6].
for hundreds of years, and their clinical and Dandelion improves the function of liver,
pharmacological effects have been extensively pancreas and stomach. It is used to treat
studied from various viewpoints [1].This anemia, cirrhosis of the liver, hepatitis and
study focused on a used in folk medicine, rheumatism [1,7] anti-inflammatory, anti-
Taraxacum officinale, commonly called oxidative, anti-carcinogenic, analgesic, anti-
Dandelion, which is a herbaceous perennial hyperglycemic, anti-coagulatory and prebiotic
plant of the family Asteraceae (Compositae). effects [8-13].
It can be found growing intemperate regions of The leaves can be eaten cooked or raw in
the world, in lawns, on roadsides, on disturbed various forms, for assembling salads or soup,
banks and shores of water ways and other which also recommended as a natural
areas with moist soils. Dandelion is considered source of vitamin C in the early spring.
a weedy species [2]. It has many english Dandelion water extract has anti-tumor
common names including: Blowball, Lion's- activity attributed to polysaccharide [1].The
tooth, Cankerwort, Swine's snout, & etc [3], most important biologically active components
Arabic names :Hindiba, Khas berri [4], The are sesquiterpenic lactones, biotin, inositol
genus name Taraxacum ,might be from the and vitamins B, D, E and phosphorous (P).
Arabic word "Tharakhchakon" [2], or from the The leaves possesses a higher content of
Greek word "Tarraxos" .The common name ²-Carotene than carrot and more Fe and Ca
"Dandelion" comes from the French phrase level than spinach, along with macro- and
"Dent de lion" which means "Lion's tooth", in micro- elements [1,14].
references to the jagged shaped foliage [5]. Flavonoids and coumaric acid derivatives
Dandelion serves mainly as a diureticum, and were identified from dandelion flower [7].Its
7
 Abdul Kadir M. N. Jassim
roots have been used to make a coffee like The pH of the extract was determined using
drink and the plant had used by Native pH-meter (Orion,SA720).
Americans as a food and medicine [15].
b) Alcoholic extract:
However, its use has mainly been
Prepared as in water extract described
based on empirical finding .This contribution
above, but with using ethanol 70% (v/v)
provides a comprehensive review of the
instead of water to give alcoholic extract
pharmacologically relevant compounds of
powder [16-19]. The pH of this extract was
Taraxacum officinale characterized so far and
also determined.
of the studies supporting its use as a medicinal
[C] Determination of Ash content:
plant.
Dried leaves 2 g were taken and heated at
Accordingly, the leaves of Taraxacum
900ºC for 20 min. in muffle furnace until the
officinale has drawn the attention of researches
material converted to white powder, after its
and consumers and an understanding of the
cooling the percentage of ash content was
chemical and pharmacological properties
determined [20].
should be of importance from scientific points
[D]Chemical detection of the plant
of view, therefore, this study is aimed at
components:
evaluating the ash contents, pH, leaves
The chemical components of the prepared
components, trace elements, effect of its
watery and alcoholic extract were detected as
extracts on different microorganisms and
shown in Table (1). They included: glycosides,
enzyme,checking of some the presence of
alkaloids, saponins, phenolic compounds,
flavonoids in these extracts.
tannins, resins, flavonoids [16-18] and proteins
Materials and Methods [21].
Protocol of Method
[E] Determination of trace element levels:
[A] Collection and treatment of samples:
Dried leaves 3 g were taken and mixed
The leaves of Taraxacum officinale were
with 8 ml of concentrated nitric acid and 2 ml
collected from the north of Baghdad, Iraq.The
of 60% prechloric acid in a conical flask, the
leaves were transported to the laboratory,
mixture was kept for 24hrs covered with watch
washed with water, cleaned with filter paper or
glass. After that it was left for 6hrs at sand
soft clothes to remove all traces of dust and
bath at 80ºC, until the digestive material
insects, then dried in shad 25-30ºC for one
converted to white powder. Deionized water
week, with continuous overturn to prevent
8 ml were added to this powder, and the
mould. weighed, ground in a mortar and
trace elements were determined [20] by
pestle, placed in airtight bottles and stored in
(Shimadzu AA-670, Flame Atomic Absorption
dissicator to by used for extraction.
Spectrophotometer).
[B] Preparation of extracts:
[F] Inhibition of hyaluronidase:
a) Watery extract:
Hyaluronidase inhibition activity was
Air dried leaves 50 g were suspended in 1
determined turbidimetrically by the method of
liter of distilled water and left for 24 hrs at
Kass et al [22] by using 0.01 mg/ml enzyme
35ºC with continuous stirring in shaking
mixed with 250 µg/ml from the extracts with
incubator. Then the mixture was filtered by
inhibition time 45 min. and the percentage of
filter paper. The filtrate was centrifuged for 10
inhibition %I was calculated according to this
min. at 2500 rpm, and the extract was
equation [23]:
evaporated to dryness at 40ºC in the incubator.
Activityof control - Activityin the presence of Extract
% Inhibition =ð ´ð100
Activityof control
[G]The biological activity: isolates were obtained from department of
The biological activity against various Biology/College of Science /Al-mustansiriyah
bacterial species was determined. As gram University .The concentrations used for each
negative bacteria, Proteus mirabilis and E.cole extracts were 0.1, 0.5 and 1 mg/ml [18, 19].
were used, while Staphylococcus aureus
was used as gram positive bacteria. These
8
Journal of Al-Nahrain University Vol.15 (3), September, 2012, pp.7-14 Science
[H] HPLC Analysis: Results and Discussion
Analysis of flavonoids, Kaempferol and Our results showd that pH value for watery
Morin, were carried out in the chemical and alcoholic extracts are pH =8.15 and 4.81,
research center, ministry of science and respectively (This may reflect the presence
technology, Baghdad, Iraq, by using of alkaloids in the watery extracts). The
(Shimadzu, LC2010A, Japan) HPLC. Standard Ash content for the Taraxacum officinale
solution were prepared (standard Kaempferol leaves is (12.8%). The qualitative chemical
and Morin were gift supplied from analysis of the watery and alcoholic extracts
Dr.Mohammed Mustafa Radi). are represented on Table 1,which show that
A luna 5U C-18Column(250mm x 4.6 x leaves contents are (glycosides, alkaloids,
5µm)Was used and coupled with 20µl of phenolic compounds, tannins, flavonoids and
sample at 40ºC with a linear gradient mobile proteins) in watery extract and the absence of
phase containing solvent A (water), solvent B alkaloids in the alcoholic extract. Other studie
(acetonitrile) and solvent C (5% formic acid show that the major phenolic fraction in
in water, v/v) with flow rate set at 0.2ml/min. dandelion is anthocyanins, a type of flavonoids
The linear gradient program started with [1,14], phenolic acid [25] and poly phenolic
88% A:10% B:2% C (v/v) and finished at 73% compounds [26], these phenolic compound
A:25% B:2% C (v/v) with retention time are an ecologically significant class of
10min. The chromatograms were recorded at secondary metabolites expressed in all higher
280nm. Standard and leaves extracts samples plants with high chemopreventive potential
were made at 25ppm then analyzed directly by [27]. The saponins and resins are not found in
HPLC [24]. both extracts.
Table (1)
Chemical components analysis (qualitative methods) for watery and alcoholic extracts of
Taraxacum officinale leaves.
Result Result
Components Reagents Note Watery Alcoholic
extract extract
Iodine test Brown ppt.
Ve+ Ve-
Glycosides Molish test Violet ring Ve+ Ve+
Ve+ +Ve
Benedict test Orange ppt.
Mayer's reagent No white ppt.
Ve- Ve-
Alkaloids Wagner reagent Brown ppt. Yellow Ve- Ve-
Ve- Ve-
Picric acid ppt.
No dense foam for
Fast stirring Ve - Ve-
Saponins long time
Mercuric Chloride Ve - Ve-
No White ppt.
Phenolic Aqueous%1 Green ppt.
Ve+ Ve+
compounds Ferric chloride
Aqueous%1
Green ppt. Ve+ Ve+
Tannins Ferric chloride
Preface yellow ppt. Ve+ Ve+
Lead acetate%1
Ethanol + Boiling +
Resins No turbidity Ve- Ve-
Distilled Water
aqueous%1
Ferric chloride Green ppt. Ve+ Ve+
Flavonoids
Ethanol hydroxide Yellow ppt. Ve+ Ve+
alcohol
Proteins Folin-Ciocalteau reagent Blue color Ve+ Ve+
9
 Abdul Kadir M. N. Jassim
The concentration trace elements in This work shows, the two extracts were
Taraxacum officinale leaves are represented in examined for their effects on hyaluronidase.
Table (2) which shows, high concentrations The percentage of inhibition for each extract
of (K, Ca, Na, Fe) with (185.1, 22, 19.5, 11.2) was 1.3% with respect to control assays run
ppm, respectively, and low concentrations simultaneously. Kuppusamy, et al. show that
of (Zn, Cd, Cu) with (6.3, 1.3, 0.2) ppm, morin and kaempferol, types of flavonoids,
respectively, other reports indicated the having potent inhibitory effect on this enzyme
presence of these metal in this plant by using [30] with 56%, 31%, respectively. Absence of
different method [28,29], while (Cr, pb) were these two flavonoids, as shown latter by
not found. The high concentration of HPLC, may cause the lower inhibitory effects
potassium cause the leaf acts as a diuretic; this of these extracts on hyaluronidase.
spares the human body's potassium, which The effect of these extracts on different
tends to be excreted with diuretic use. The microorganisms were studied and compared.
effect is more therapeutic and beneficial However, the results in Table (3), show that
overall. The young leaves that come up in the the concentrations 0.5 and 1 mg/ml exhibit
spring may also be add to spring salads for this effective inhibition towards the growth of
benefit [14]. Regardless of its intended use, the intended bacteria for both extracts specially on
high mineral content of this herb greatly Gram positive, Staphylocoous aureu, while
increases the chance of drug-drug interactions less inhibition effects were seen for Gram
with the conventional medicines that are negative, Proteus mirabilis and E.coli.
sensitive to cations [13]. Alcoholic extract with concentrations
0.5 mg/ml was more effective inhibitor for the
Table (2)
Gram negative, E.coli than that for the
The concentration trace elements content of
watery extract; and the concentration
Taraxacum officinale leaves.
0.1 mg/ml of this extract failed to inhibit any
Concentration
microorganisms.
Trace elements symbol
(ppm)
Potassium K 185.1
Calcium Ca 22
Sodium Na 19.5
Iron Fe 11.2
Zinc Zn 6.3
Cadmium Cd 1.3
Cupper Cu 0.2
Chrome Cr Nil
Lead Pb Nil
Table (3)
The effect of watery and alcoholic extracts of Taraxacum officinale represented by inhibition
zone (mm) against different bacteria species.
Bacterial species Alcoholic extract (mg/ml) Watery extract (mg/ml)
0.1 0.5 1 0.1 0.5 1
Staphylococcus
++ ++ ++ ++
- -
aureus (gram + ve)
Proteus mirabilis
+ + + +
- -
(gram - ve)
E.coli
+ + +
- - -
(gram - ve)
(-) No inhibition zone
(+) Inhibition zone between (1-4) mm .
(++) Inhibition zone between (4-10) mm .
01
Journal of Al-Nahrain University Vol.15 (3), September, 2012, pp.7-14 Science
This inhibitory effects may be duo to the The HPLC chromatogram in Fig.(1),
(glycosides and/or phenolic compounds and/or shows the standard Morin (A), standard
tannins and/or flavonoids and/or alkaloids Kaempferol (B) and leaves extract sample (C).
and/or proteins) presence in the plant extracts. Identification of these flavonoids in leaves
Such compounds had been reported to have an extract sample was checked according to
active effect on the bacterial cells membrane, retention time obtained from standard run at
which may caused destroy these identical conditions.
microorganisms [7, 18]. The alkaloids interact The major peaks for Morin standard had
with the DNA, the tannins inhibit the carrier retention time was 2.35min (Fig.(1, A)), while
enzymes and proteins present in the cells for that of Kaempferol was 3.3 and 3.56 min.
membrane, while the phenolic compounds (Fig.(1, B)). As it is clear from the
form complex with dissolved protein out of the chromatogram. These two flavonoids were
cells or with cells membrane which made to absent in the leaves of Taraxacum officinale.
destroy the bacteria [18, 22].
HO
1
O
OH
7 B
HO
A C
3
OH
5
O
HO
(A) Standard Morin
OH
1 B
O
7
HO
C
A
3
OH
5
O
HO
(B) Standard Kaempferol
(C) leaves watery extract
Taraxacum Officinale
Fig.(1) HPLC chromatogram of (A) standard Morin, (B) standard Kaempferol, (C) leaves watery
extract for Taraxacum officinale leaves.
Using luna 5U C-18Column(250mm x 4.6 x 5µm) the mobile phase at 40 ºC containing solvent A
(water) ,solvent B (acetonitrile) and solvent C (5% formic acid in water ,v/v) with flow rate set at
0.2ml/min. The absorbance at (280nm).
00
 Abdul Kadir M. N. Jassim
Conclusion [8] chutz, k.; Carle, R. and Schieber, A.,
The present study confirm that plant leaves "Taraxacum A review on Its phytochemical
extracts (watery and alcoholic) for Dendelion, and Pharmacological Profile, Journal of
Taraxacum officinale leaves posses in vitro Ethnopharmacology", 107,3, 2006, 313-323.
antibacterial activity because of its content,
[9] Shi, S.; Zhao, Y.; Zhang, Y.; Huang, K.
glycosides, alkaloids, phenolic compounds,
and Liu, S., "Phenylpropanoids From
tannins, flavonoids, proteins and trace
Taraxacum mongolicum", Biochemcal
elements, however, if plant leaves extracts are
Systematics and Ecology, 36, 2008, 716-
to be used for food preservation or medical
718.
purposes, issues of safety and toxicity will
[10] Shi, S.; Zhao, Y.; Zhao, H.; Zhang, Y.;
need to be addressed, and this results will
Jiang, X. and Huang, K.,". Identification
serve as a pilot experiment for further research
Antioxidants From Taraxacum mongolicum
and improvement strategies of this important
by High-performance Liquid Chromato
plant .
raphy  Diode Array Detection  Radical -
Scavenging Detection - Electrospray
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02
 Abdul Kadir M. N. Jassim
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03