‫الرحيم‬ ‫الرحمن‬ ‫هللا‬ ‫بسم‬
Al-Quds University
Faculty of Science & Technology
Applied and Industrial Technology Program
Extract of Stinging Nettle, Phase behavior and
total bioactive compounds
Prepared by :Afaf Abuhilal
Registration No:2012450
Supervisor: Dr. Ibrahim Kayyali
Co Supervisor: Dr. Fuad Al Rimawi

Introduction
• stinging nettle, (Urtica dioica), common nettle,
weedy perennial plant of the nettle family
(Urticaceae).
• The stinging trichomes of the leaves and stems
have bulbous tips that break off when brushed
against, revealing needlelike tubes that pierce
the skin.
• It is a wild plant, In Palestine we find it in
colonies, common (
‫قريص‬
) it is known for years in
medicine and kitchen use.

Nutritional composition:
-Fresh leaves contain approximately: 90% moisture,
up to 3.7% proteins, 0.6% fat, 2.1% ash, 6.4%
dietary fiber and 7.1% carbohydrates .
-sterols , fatty acids, polysaccharides,
-minerals such as(iron ,potassium ,manganese ,
calcium.)
-vitamin(A, B, C, D and K).
-Protein and amino acids
-Poly phenols (Ursolic acid and quercetine).

Polyphenols of Urtica
Phenolic compounds from U. dioica roots (9)

Medical benefits of Urtica:
Graphical abstract (8)

Medical Benefits of Urtica
• It inhibits Corona virus replication, a study was
conducted on a mouse model. (10)
• When Treated with UDA at 5 mg/kg significantly
protected the mice against a lethal infection with
mouse-adapted SARS-CoV (p<0.001.
• The mechanism of Action of Urtica, it might bind
to Nacetylglucosamine-like residues present on
the glycosylated envelope glycoproteins, thereby
preventing virus attachment to cells.

Side effects and toxicity:
• It is considered as a safe plant.
• Internal use of Nettle might cause diarrhea
, constipation, and upset stomach in some
people.
• When applied to the skin: Stinging nettle is
possibly safe. Touching the stinging nettle plant
can cause skin irritation.
• Fresh nettle causes stings and can rarely lead to
severe allergic reactions in susceptible
individuals.

Preparation of plant material:
2- dry the plant for 8-10 days
4- 150 ml ethanol 5% in soxhlet
8 hour
3- crushed into powder
5- evaporator using rotary
evaporator
1- collected nettle
plant (leaves, stalk &
(
roots

Methods of Extraction:
Extraction: is the separation of medicinally active
portions of plant or animal tissues from inactive
or inert components by selective solvents in
standard extraction procedures.
-method of extraction:
1-Soxhlet Extraction.
2-Ultrasound-assisted Extraction (UAE)
3-Supercritical Fluid Extraction (SFE)
4-Shake extraction
5-Accelerated Solvent Extraction (ASE)

Soxhlet Extraction:
In this method ,the finely solid material containing
desired compound is placed inside thimble made from
thick filter paper,which is loaded into the main
chamber of the extractor. The extracting solvent in
flask is heated and its vapors condense .
The condensed extractant drips into the thimble
containing the crude drug and extract it by
contact,when the level of liquid in chamber rises to the
top of siphon tube ,the chamber automatically empty
by siphon side arm into flask.

Method for phenolic acid
determination:
For separation ,purification and identification of
phenolic compound ,numerous
chromatography method have been used
,such as:
1-High performance liquid chromatography
(HPLC).
•
2-Gas chromatography
•
3-Capillary electrophoresis

Total content of Phenols
• Folin-Ciocalteuassay involves the reduction of
the F–C reagent with phenolic compounds in an
alkaline medium. The reaction is accompanied by
the formation of a blue-colored complex that has
a maximum absorbance at 765 nm. The
absorbance is directly proportional to the TPC
which is reported as Gallic acid equivalent.
• Quantitation of total flavonoid content (TFC) is
widely performed using the aluminum chloride
colorimetric assay against a flavonoid standard
assuming equal responses from all flavonoids

Antimicrobial activity
• The antimicrobial effects of the plant extracts will be evaluated
using Antimicrobial tests which will be carried out by the Disc
diffusion method.
• This method is based on the diffusion of an antibiotic from a filter
paper disc through the solidified culture medium (Muller- Hinton
agar) of Petri dishes.
• Whatman filter paper will be used to prepare small discs (6 mm in
diameter), then sterilized at 121 C for 15 min in an autoclave. The
discs were separately impregnated with the extracts for 1-4 hour at
room temperature and allowed to dry.
• A swap of bacterial broth will be dispersed on the agar plate
surface. After drying, the previous discs placed on the Petri dishes
and then incubated at 37 C for 24 to 48 hour. Three replicated
plates used for each treatment. The diameter of inhibition zone
created by each disc will be measured (in mm) using a ruler.

Literature review:
• (Kornelia Koszegi, etal, 2014)) applied three kinds of
solvents into two type of extraction techniques :
• Soxhelt extraction with 96%ethanol,Soxhelt extraction
with n-hexane and supercritical fluid extraction
(SFE)with CO2,by repeating the measurement and
analysis three times,
• The result exhibited that Soxhlet extraction with
ethanol is the highest while the soxhlet extraction and
the super critical extraction has lower yield ,the total
polyphenol content and the H-donating ability of these
extracts were low .

Literature review
• (Semih Otles and Buke, (2011)), different amount of
nettle were collected from different region to study its
phenolic component and antioxidant activities , nettle
is seperated into 3 part:leaves,stalk and leaves.
• To analysis them he used HPLC .
• Folin –Ciocalteu method to collect total phenolic
components ,and DPPH to measure antioxidat activity.
• According to the result the component of each part
had different quantity and quality for example phenolic
content :
• Root 7.82/stalk 9.91/leaves 7.62gm

Literature review
• ( Hanan Bander et al,(2013)),studied and applied different
method and techniques with different solvent for
extraction of bioactive compound from Nettle
.(maceration,reflux,soxhlet,hydrodistillation,Ultrasound-
Assisted Extraction (UAE)and Microwave-Assisted
Extraction (MAE)).
• Using different solvents (hexane,dichloro
methane,aceton,ethanol,water).
• The result show that the extraction method , solvent and
time had a significant effect on the amount of extracted
compound .Ethanol was the most effective organic solvent
and soxhelt had the highest extraction yield.

Literature review
• Namazi et al,(2011),studied the effect of nettle extract on
insulin sensetivity and some inflammatory indicators in
type2 diabetic patient ,this was randomized double-blind
clinical trial on 50men and woman with type2 diabetes
done over an 8 week period .
• They received 100mg nettle extract in3 portions a day for
8weeks.interleukin6 (IL-6)and Fasting insulin concentration
were measured .
• Insulin sensitivity was calculated at the beginning and the
end of the study .
• After8 weeks, IL-6 showed a significant decrease ,so this
finding showed that nettle extraction has decreasing effect
on IL-6 in patients with type2 diabetes after 8 week trial.

Literature review
• (Capucine Bourgeois et al(2016)),used HPLC
method to determined ursolic acid and quercetin
in nettle extract, and to evaluated the antioxidant
and anti-aging activity of these extract.
• The anti-aging potential involves the inhibition of
enzyme activity such as elastase and collagenase,
and these inhibitory effect due to ursolic acid and
quercetin present in nettle extract .
• The result also show the extraction parameter
such as :extraction time,temperature,and ethanol
percentage.

Literature review
( Fedorovska Maryana Ivanivna et ,al,(2018)).
Used nettle fresh upper parts of stem with 5-6
leaves to developed gel with nettle juice and
study its effect in rats .
The result show that increase in the growth of
rat wool ,increase in the number of the mast
cell and increase in the hair follicle .

Construction of phase diagram:
-The pseudo ternary phase diagram consisting
of oil, water, surfactant and co-surfactant
mixture of different HLB values were
constructed using water water titration
method .
1-The sample of extracted oil and nonionic
surfactant are insert into 10ml glass test tube
with screw caps at room temperature at
different weight ratios.

Construction of phase diagram
2-mechenical shaking for 1min
3-titration with water is injected into each test tube
(drop by drop).
4-vigorous mechanical shaking for 5min ,on the
vortex due to the high viscosity of the surfactant
and oil in order to guarantee homogenous
dispersion .
5-placing the tube within thermostatic water bath
to control the temperature.
6- To reach equilibrium left rest for 24hrs then the
addition water and analyzing the samples.

Construction of phase diagram
9:1
8:2
7:3
6:4
5:5
4:6
3:7
2:8
1:9
Weight ratio
0:9
0:8
0:7
0:6
0:5
0:4
0:3
0:2
0:1
Surfactant
1:0
2:0
3:0
4:0
5:0
6:0
7:0
8:0
9:0
Oil

Determination of phase behavior :
1-Visual observation: microemulsion will
identify by color, texture and transparency.
2- Anisotropy :is detected by the cross
polarizer.clear isotropic,one-phase system
were designated as microemulsion system .

Time table:
July
2023
May
2023
March-
APR
2023
Feb
2023
Dec-Jan
2022
Nov
2022
X
X
1- Literature Review
X
2-Collecting and
preparing sample
X
X
X
3-Extraction
X
4- writing
X
5-Presentation of
results

References:
1-Ilhami Gulcin , O. Irfan Kufrevioglu , Munir Oktay ,Mehmet Emin Buyukokuroglu,
Antioxidant, Antimicrobial,antiulcer and analgesic activities of nettle ,Journal of
Ethnopharmacology (2004) 205-2015.
Kornelia koszegi, Gyula Vatia , Erika Bekassy-Molnar, Comparison the Soxhlet and
Supercritical Fluid Extraction of Nettle Root, Periodica polytechnica Chemical
Engineering, (2015) 168-173.
3-Solvita Zeipina,InaAlsina,Liga Lipese and Mara Duma . Antioxidant Activity in Nettle
and Garden Orache (Atriplex Hortensis L) Leaves During Vegetation Period
.Chemine Technologija (2015) 28-33.
4-Semih Otles and Buket Yalcin, Phenolic Compound Analysis of Root,Stalk and Leaves
of Nettle.The scientific World Journal, 12pages (2012).
5-Sadegh Fattahi ,Ebrahim Zabih,Zeinab Abedian, Roghayeh Pourbagher ,Ali
Motevalizadeh Ardekani, Amrollah Mostafazadeh and Haleh Akhavan-Niaki.Total
Phenolic and Flavonoid Content of Aqueous Extract of Stinging Nettle and in Vitro
Antiproliferative Effect on Hela and BT-474 Cell Lines.Int Jmol Med Spring (2014) 1-

References
6-Capucin Bourgeois, Emilie A.Leclerc ,Cyrielle Corbin, Joel Doussot,Valerie Serrano,
Jean-Raymond Vanier,Daniel Auguin, Eric Lain and Christoph Hano. Nettle as
asource of antioxidant and anti-aging phytochemicals for cosmetic application
,Comptes Rendus Chemie(2016)1090-1100.
7-Lenka Drostinova, Jana Branisa, Dominika Boncikova and Klaudia Jomova. Effect of
Drying Methods on Content of some Natural Pigments in Urtica DioicaL. An
Melissa Officinalis L. Journal of Microbiology, Biotechnology and Food
SciencemScience,(2015)182-185.
8-Stinging Nettle (Urtica dioica L.): Nutritional Composition, Bioactive Compounds,
and Food Functional Properties by Hari Prasad Devkota, Molecules 2022, 27(16),
5219
9-Therapeutic Perspectives of Molecules from Urtica dioica Extracts for Cancer
treatment by Sabrina Esposito Molecules 2019,24 (15), 2753
10- Yohichi Kumakia, Miles K. Wanderseea, Aaron J. Smitha, Yanchen Zhoub.Inhibition
of severe acute respiratory syndrome coronavirus replication in a lethal SARS-CoV
BALB/c mouse model by stinging nettle lectin, Urtica dioica agglutinin.J.Antiviral
(2011) 90(1): 22–32.