SlideShare a Scribd company logo
1 of 38
B.PHARM. 5th SEMESTER
PHARMACOGNOSY ANDPHYTOCHEMISTRY-II
BP504T.
UNIT-III
Isolation, Identification and Analysis of
Phytoconstituents
Terpenoids: Menthol, Citral, Artemisin
What is terpenoids
Any of a large class of organic compounds including
terpenes, diterpenes, and sesquiterpenes.
They have unsaturated molecules composed of linked
isoprene units, generally having the formula (C5H8) n.
Isoprene unit are 5 carbon unit
For having isoprene unit they are also called
isoprenoids
Physical properties of terpenoids
Odorous
Volatile at room temperature
They represents essence or active constituents of plants hence
called essential oil
Lighter than water
Have high refractive index
Are optically active
Chemical Properties of teprenoids
Terpenoids are the hydrocarbones of plant origin of the general formula (C5H8)n
as well as their oxygenated, hydrogenated & dehydrogenated derivatives
Terpenoids are regarded as derivatives of polymers of isoprene, CH3H2C=C-
CH=CH2 (i.e.C5H80)
They consist of a complex mixture of terpenes or sesquiterpenes, alcohols,
aldehydes, ketone, acid & esters.
They are unsaturated compounds.
They undergo addition reaction with hydrogen, halogen, halogen acids to form
addition products.
They undergo polymerization and dehydrogenation in the ring.
On thermal decomposition, terpenoid gives isoprene as one of the product.
Chemical Properties of teprenoids
Terpinoids are modified terpenes, where in methyl groups
have been moved or removed, or oxygen atoms added.
Sometimes terpinoids are added to proteins, e.g., to
enhance their attachment to the cell membrane.
The steroids and sterols in animals are biologically
produced from terpinoid precursors.
MENTHOL
CH3
CH3
H3C
OH
Menthol
• Biological source: Menthol a monoterpene alcohol
obtained from oils of Mentha piperita var. Vulgaris
(Black peppermint) & Mentha piperita var. officinalis
(White peppermint),
• Family: Labiatae.
Mentha species contains about 1-3% of volatile oils & oil
contains not less than 44%.
G.S.: Asia, Europe, North America, Cultivated in India,
japan, Germany, France, Italy.
Menthol is isolated by 2 methods
Isolation
Method 1: Hydro-
distilation
Method 2: Steam
distillation
Hydro-distillation
Required quantity of coarse powder leaves
of Mentha piperita.
The peppermint oil is extracted by
hydrodistillation method by using clevenger
apparatus.
The oil is separated from water and allow to
cooling. After cooling crystals of (-) menthol
will separate out.
The crystals are collected by centrifugation
and re- crystallized the menthol by acetone
or any other low boiling point solvent.
1. Hydro-distillation by Clevenger apparatus system
2. Steam distillation
Required quantity of air dried mentha
plant is charged into stainless steel still
having perforated bottom.
The steam under pressure is generated
with the help boiler and steam is passed
through the drug.
The mixture vapour (water and volatile oil) are
passed through the condenser where vapour is
cooled and back to liquid form.
The mentha oil is collected in separating
can. Mentha oil is floated on top of the
water due to lighter than water.
The oil is then decanted and filtered.
2. Steam distillation
3. Method of Isolation
Pippermint volatile oil- separated by steam & water distillation.
To make oil free of moisture- it is passed through a bed of Na2SO4
The moisture free oil is frozen to -60°C for 7days by packing in tightly sealed plastic
container.
Menthol in oil separates as flaky crystals, separated by filtration.
Mother liq. Containing some menthol along with other terpenes.
The mother liq is treated with boric acid & further boiled for 3hr, followed by distillation
for removal of menthone.
Borate of menthol subjected to saponification with 50ml of NaOH by heating reflux for
1hr
Resultant sol. Is allowed to cool for separation of remaining menthol crystals and mixed,
dried in dessicator.
Properties:
• Appearance: White crystalline substances, which is solid at room
temperature and melts slightly above (m.p. 41 to 43°C).
• Odor: Characteristic and pleasant
• Taste: Pungent followed by cooling sensation
• Solubility: Soluble in 70% alcohol, ether and chloroform, insoluble
in water
• Identification by chemical test:
• Few drops of sample is mixed with 5ml of nitric acid and heated on
water bath. Blue color is developed within 5 minutes, after some
time it becomes yellow which indicate the presence of menthol.
• Small qty of menthol in TT + equal qty of thymol or camphor →
liquifaction of contents in TT→ presence of Menthol
• Crystals of Menthol in watch glass+ heat on water bath → material
evaporates
Analysis
Take 10gm drug, add 10ml of acetic anhydride & 2gm of sodium
acetate to the flask. Reflux for 1 hr.
After cooling add 30ml water, boil it for 15 min. with continuous
stirring. Then transfer in separating funnel.
The oil layer is washed with water until it become neutral.
Add 2 gm of sodium sulphate & shake it. Allow it for 30 min. &
filter it.
Take both layer in different flask, add 5 ml ethanol and
Phenolphthelin indicator by adding 25 ml KOH ethanol solution.
(for neutralize it)
Again reflux for 1 hr. cool it titrated with HCl.
• % Menthol = 7.814 x (c-a) {0.021 x (c-b)/U}
A– 0.021 x (c-a)
• Where,
“a” & “b” = ml of HCl for oil
“c” = ml of HCl for blank
“U”= Un-acetylated oil
“A”=Acetylated oil
Analysis by TLC
• Sample preparation: 1mg of Menthol is dissolved 1ml of methanol
• Standard sample: Menthol
• Stationary phase: Silica gel 60 F 254
• Mobile phase: Pure Chloroform OR hexane: EtOAc :: 8: 2
• Detecting agent:
1% vanillin-sulphuric acid reagent and heat the plate at
110°C for 10 minutes
Anisaldehyde-sulphuric acid reagent, heat at 115°C-10min.
• RF Value: 0.34, 0.48-0.62
Utilization
• It is used as in various dosage forms for its cooling sensation,
flavoring property,
• carminative, antispasmodic and antipruritic
Storage condition
• It should be store in well closed and air-tight containers protected
from light and in cool place.
CITRAL
Citral
Biological source: Citral is a monoterpene aldehyde obtained
from oils of lemon grass Cymbopogon flexuosus and C. martini.
Family: Graminae.
It also present in Lemon oil (Citrus limonus) and Orange oil
(Citrus aurantium).
Family: Rutaceae.
Lemon oil: Lemony, sweet smell, yellow in color.
It contains not less than 75% of aldehyde calculated as Citral.
Citral is isolated by following methods
Isolation
Method 1: Hydro-
distillation
Method 2: Fractional
crystallization Method
Hydro-distillation
Required quantity of coarse powder leaves of
lemon grass or other powder.
The lemon grass oil is extracted by hydro
distillation method by using Clevenger
apparatus.
The oil is separated from water and allow to
cooling. After cooling of lemon grass will
separate out.
Citral
The fresh plant material is hydro-distilled
to obtain lemon grass oil.
It is purified by fractional crystallization.
To the total oil, first Sodium sulphite is added,
the citrals get converted into its sulphite salt.
The salt crystallizes out of the solution.
The crystals are filtered and washed with
ether or chloroform.
Properties:
• Appearance: Clear pale yellow liquid
• Odor: Strong lemon like odour
• Taste: Lemon like taste
• Solubility: Soluble in 3 parts of 70% alcohol,
chloroform and fixed oil. Insoluble in water.
• Boiling point: 224-228 0C
• Identification by chemical test:
• Alcoholic solution of Sudan red III is added to the
sample. Red colour is appeared which indicate
presence of citral.
• Tincture alkane is added to the sample. Red colour
is appeared which indicate presence of citral.
• Analysis by TLC
• Sample preparation : 1mg of Citral is dissolved 1ml of methanol
• Standard sample : Citral
• Stationary phase : Silica gel –G
• Mobile phase : Pure Chloroform
• Detecting agent : 2, 4, dinitrophenyl hydrazine reagent
• Color spots : Yellow to orange
• RF Value : 0.51
• Utilization:
• It is used as a flavouring agent and perfumery. Commercially citral
is act as precursor for the synthesis of ß-ionone. ß-ionone is used
as starting material for the synthesis of Vitamin A
• Storage condition:
• It should be store in well closed and air-tight containers protected
from light and in cool place.
ARTEMISIN
Artemisin
Biological source: Artemisin is a sesquiterpenoid
lactone, obtained from the unexpanded flower- heads of
Artemisia cina Berg, Artemisia brevifolia Wall,
Artemisia maritime Linn. and other species
Family: Asteraceae.
It contains 2-3% of essential oil and two crystalline
substances i.e Santonin and Artemisin.
G.S: It is found in Italy, Iran, South Austrelia, In India
Gujarat, J&K, Karnataka, UP and Hariyana
Extraction
Extraction can be done
by 2 ways;-
Soxhlet extraction
Microwave-assisted extraction
REQUIREMENTS
• Raw material
• Solvents (methanol, n-hexane, n-heptane,
benzene, toluene, chloroform, methylene
dichloride, ethyl acetate, acetone, acetonitrile..)
SOXHLET EXTRACTION AND TEST SAMPLE
PREPARATION
100 mg of fine powder was placed into an extraction
thimble
extracted with 170 ml of solvent via hot soxhlet extraction
method for 6 hours over a water bath
The extract was evaporated and redissolved in 5 ml
methanol
10 μl of these test solutions was used for quantification
purpose.
MICROWAVE-ASSISTED EXTRACTION AND TEST
SAMPLE PREPARATION
10 μl of these test solutions was used for quantification purpose.
The extract thus obtained was evaporated and redissolved in 5 ml methanol
For microwave-assisted extraction (MAE) the same parameters should be followed for every
solvent
Extraction parameters (160 watts, 120s, 10 ml per extraction cycle, two extraction cycles,
and cleanup with 2 ml of corresponding solvent at the end of second cycle of extraction)
100 mg of fine powder was extracted under the influence of microwave energy using
solvents
Partition the extract using 50 ml hexane
Partitioning done many times until hexane layer become colorless
Two layers got from this process
The methanol extract obtained was added 10 ml of distilled water
Partitioned again using 50 ml of ethyl acetate
Partitioning done many times until the ethyl acetate layer is colourless.
Again we get 2 layers
PARTITION OF EXTRACT
1. hexane extracts (non-
polar fraction)
2. methanol extracts
Each extract was concentrated using a rotavapor at a temperature of 40˚c.
2.methanol-water
extract (polar fraction)
1.ethyl acetate extract
(semipolar fraction)
ESTIMATIONS
By 2 methods:-
• TLC Densitometer
• HPTLC method
TLC METHOD
• Mobile phase :ethyl Acetate: hexane (3:97)
• Stationary phase :60 F254 silica gel
• Detecting reagent :anisaldehyde-sulphuric acid reagent
• Spot volume :10 μL of test and standard sample spots
• Spot colour :Pink-colored spots of artemisinin
• Room temp. :25 +- 2˚C
• Relative humidity :45+- 2%
Extrapolating the graph to x axis give
concentration of test sample
Values are plotted in calibration curve (con
Vs spot area)
A simple TLC-densitometric technique has
been developed for the rapid and accurate
analysis of artemisinin in a at 540-610 nm
using tungsten lamp
large number of Artemisia annua plantlets
HPLC METHOD
•EQUIPMENT MODEL: -- WATERS 501
•COLUMN: -- WATERS C18
•DETECTOR: -- UV 260 nm
•MOBILE PHASE: -- phosphate buffer : methanol (6:4) – PH – 7.9
•FLOW RATE: -- 1ml/min
Calibration curve was plotted with diff conc
Description - - colour less needles /white crystalline powder,
odourless
Identification - - TLC/ HPLC with std
Molecular formula - - C15H22O5
Molecular wt - - 282.3
Solubility - - partially insoluble in water
very soluble in dichloromethane
MP - -
freely soluble methanol, ethanol
151-154 c
LOSS ON DRYING - - NMT 0.5%
STORAGE - - Tightly closed container
SPECIFIC SHEET
Identification by chemical test:
Sample is boiled with 10ml of alcohol and filtered. Sodium
hydroxide is added to filtrate and heated. Red colour is
appeared in liquid .
• Analysis by TLC
• Sample preparation: 1mg of Artemisin is dissolved
Chloroform
• Standard sample: Artemisin
• Stationary phase: Silica gel –G
• Mobile phase: Petroleum ether-Ethyl acetate (1:2)
• Detecting agent: p-dimethyl amino benzaldehyde and
heat at 80° C to produce color
• RF Value: Compare with standard Artemisin
Artemisnin
• Analysis by UV
• 1mg of Artemisin is mixed with 10ml of methanol
and analysed λ200-400nm.
• Utilization:
• It is used as antimalarial.
• It acts both against chloroquine sensitive and
resistant P. falciperum and P. vivax malarial
parasites.
• Storage condition:
• It should be store in well closed and air-tight
containers protected from light and in cool place.
THANKYOU

More Related Content

Similar to 3.1 A-Terpenoids-Menthol-Citral-Artemisin.pptx

Bitter principles Lec.2 (2017)
Bitter principles Lec.2 (2017)Bitter principles Lec.2 (2017)
Bitter principles Lec.2 (2017)Ahmed Metwaly
 
Bulet kumar Gupta B.pharma 3rd year unit 4.pptx
Bulet kumar Gupta B.pharma 3rd year unit 4.pptxBulet kumar Gupta B.pharma 3rd year unit 4.pptx
Bulet kumar Gupta B.pharma 3rd year unit 4.pptxBkGupta21
 
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Srinivas college of pharmacy, Mangalore
 
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Srinivas college of pharmacy, Mangalore
 
Extraction, isolation & estimation of ephedrine
Extraction, isolation & estimation of ephedrine Extraction, isolation & estimation of ephedrine
Extraction, isolation & estimation of ephedrine KUNAL KELZARKAR
 
Terpenoids, carotenoids, vitamins and quassinoids
Terpenoids, carotenoids, vitamins and quassinoidsTerpenoids, carotenoids, vitamins and quassinoids
Terpenoids, carotenoids, vitamins and quassinoidsSana Raza
 
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptx
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptxEXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptx
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptxHeeraKaremore
 
Phytochemistry essential oils esters, oxides and peroxides
Phytochemistry essential  oils   esters, oxides and peroxidesPhytochemistry essential  oils   esters, oxides and peroxides
Phytochemistry essential oils esters, oxides and peroxidesMostafa Mahmoud Hegazy
 
Isolation, industrial production of phytoconstituents by Pooja Khanpara
Isolation, industrial production of phytoconstituents by Pooja Khanpara Isolation, industrial production of phytoconstituents by Pooja Khanpara
Isolation, industrial production of phytoconstituents by Pooja Khanpara POOJA KHANPARA
 
Phytochemistry essential oils terpene hydrocarbons
Phytochemistry essential  oils terpene hydrocarbonsPhytochemistry essential  oils terpene hydrocarbons
Phytochemistry essential oils terpene hydrocarbonsMostafa Mahmoud Hegazy
 

Similar to 3.1 A-Terpenoids-Menthol-Citral-Artemisin.pptx (20)

Isolation and identification of the phytoconstituents
Isolation and identification of the phytoconstituentsIsolation and identification of the phytoconstituents
Isolation and identification of the phytoconstituents
 
Bitter principles Lec.2 (2017)
Bitter principles Lec.2 (2017)Bitter principles Lec.2 (2017)
Bitter principles Lec.2 (2017)
 
Bulet kumar Gupta B.pharma 3rd year unit 4.pptx
Bulet kumar Gupta B.pharma 3rd year unit 4.pptxBulet kumar Gupta B.pharma 3rd year unit 4.pptx
Bulet kumar Gupta B.pharma 3rd year unit 4.pptx
 
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
 
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
Isolation,identification and analysis of phytoconstituents .Dr.U.Srinivasa, P...
 
Volatile oils ppt
Volatile oils pptVolatile oils ppt
Volatile oils ppt
 
Unit-III Pharmacognosy and Phytochemistry II.pdf
Unit-III Pharmacognosy and Phytochemistry II.pdfUnit-III Pharmacognosy and Phytochemistry II.pdf
Unit-III Pharmacognosy and Phytochemistry II.pdf
 
Extraction, isolation & estimation of ephedrine
Extraction, isolation & estimation of ephedrine Extraction, isolation & estimation of ephedrine
Extraction, isolation & estimation of ephedrine
 
volatile oil (2)
volatile oil (2)volatile oil (2)
volatile oil (2)
 
extraction of spices
extraction of spicesextraction of spices
extraction of spices
 
Terpenoids, carotenoids, vitamins and quassinoids
Terpenoids, carotenoids, vitamins and quassinoidsTerpenoids, carotenoids, vitamins and quassinoids
Terpenoids, carotenoids, vitamins and quassinoids
 
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptx
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptxEXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptx
EXTRACTION, ISOLATION & ESTIMATION OF EPHEDRINE-1.pptx
 
Phytochemistry essential oils esters, oxides and peroxides
Phytochemistry essential  oils   esters, oxides and peroxidesPhytochemistry essential  oils   esters, oxides and peroxides
Phytochemistry essential oils esters, oxides and peroxides
 
Isolation, industrial production of phytoconstituents by Pooja Khanpara
Isolation, industrial production of phytoconstituents by Pooja Khanpara Isolation, industrial production of phytoconstituents by Pooja Khanpara
Isolation, industrial production of phytoconstituents by Pooja Khanpara
 
Industrial production, estimation and Utilization of Phytoconstituents
Industrial production, estimation and Utilization of PhytoconstituentsIndustrial production, estimation and Utilization of Phytoconstituents
Industrial production, estimation and Utilization of Phytoconstituents
 
Unit-IV Pharmacognosy and phytochemistry II.pdf
Unit-IV Pharmacognosy and phytochemistry II.pdfUnit-IV Pharmacognosy and phytochemistry II.pdf
Unit-IV Pharmacognosy and phytochemistry II.pdf
 
Phytochemistry essential oils terpene hydrocarbons
Phytochemistry essential  oils terpene hydrocarbonsPhytochemistry essential  oils terpene hydrocarbons
Phytochemistry essential oils terpene hydrocarbons
 
Isolation by pooja
Isolation by poojaIsolation by pooja
Isolation by pooja
 
Fragrances
FragrancesFragrances
Fragrances
 
Volatile oils
Volatile  oilsVolatile  oils
Volatile oils
 

Recently uploaded

Narcotic and Non Narcotic Analgesic..pdf
Narcotic and Non Narcotic Analgesic..pdfNarcotic and Non Narcotic Analgesic..pdf
Narcotic and Non Narcotic Analgesic..pdfPrerana Jadhav
 
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...DhatriParmar
 
ICS2208 Lecture6 Notes for SL spaces.pdf
ICS2208 Lecture6 Notes for SL spaces.pdfICS2208 Lecture6 Notes for SL spaces.pdf
ICS2208 Lecture6 Notes for SL spaces.pdfVanessa Camilleri
 
week 1 cookery 8 fourth - quarter .pptx
week 1 cookery 8  fourth  -  quarter .pptxweek 1 cookery 8  fourth  -  quarter .pptx
week 1 cookery 8 fourth - quarter .pptxJonalynLegaspi2
 
4.11.24 Mass Incarceration and the New Jim Crow.pptx
4.11.24 Mass Incarceration and the New Jim Crow.pptx4.11.24 Mass Incarceration and the New Jim Crow.pptx
4.11.24 Mass Incarceration and the New Jim Crow.pptxmary850239
 
How to Make a Duplicate of Your Odoo 17 Database
How to Make a Duplicate of Your Odoo 17 DatabaseHow to Make a Duplicate of Your Odoo 17 Database
How to Make a Duplicate of Your Odoo 17 DatabaseCeline George
 
Unraveling Hypertext_ Analyzing Postmodern Elements in Literature.pptx
Unraveling Hypertext_ Analyzing  Postmodern Elements in  Literature.pptxUnraveling Hypertext_ Analyzing  Postmodern Elements in  Literature.pptx
Unraveling Hypertext_ Analyzing Postmodern Elements in Literature.pptxDhatriParmar
 
Active Learning Strategies (in short ALS).pdf
Active Learning Strategies (in short ALS).pdfActive Learning Strategies (in short ALS).pdf
Active Learning Strategies (in short ALS).pdfPatidar M
 
Multi Domain Alias In the Odoo 17 ERP Module
Multi Domain Alias In the Odoo 17 ERP ModuleMulti Domain Alias In the Odoo 17 ERP Module
Multi Domain Alias In the Odoo 17 ERP ModuleCeline George
 
4.16.24 21st Century Movements for Black Lives.pptx
4.16.24 21st Century Movements for Black Lives.pptx4.16.24 21st Century Movements for Black Lives.pptx
4.16.24 21st Century Movements for Black Lives.pptxmary850239
 
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptx
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptxBIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptx
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptxSayali Powar
 
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITW
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITWQ-Factor HISPOL Quiz-6th April 2024, Quiz Club NITW
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITWQuiz Club NITW
 
MS4 level being good citizen -imperative- (1) (1).pdf
MS4 level   being good citizen -imperative- (1) (1).pdfMS4 level   being good citizen -imperative- (1) (1).pdf
MS4 level being good citizen -imperative- (1) (1).pdfMr Bounab Samir
 
Scientific Writing :Research Discourse
Scientific  Writing :Research  DiscourseScientific  Writing :Research  Discourse
Scientific Writing :Research DiscourseAnita GoswamiGiri
 
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptx
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptxINTRODUCTION TO CATHOLIC CHRISTOLOGY.pptx
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptxHumphrey A Beña
 
Transaction Management in Database Management System
Transaction Management in Database Management SystemTransaction Management in Database Management System
Transaction Management in Database Management SystemChristalin Nelson
 
ROLES IN A STAGE PRODUCTION in arts.pptx
ROLES IN A STAGE PRODUCTION in arts.pptxROLES IN A STAGE PRODUCTION in arts.pptx
ROLES IN A STAGE PRODUCTION in arts.pptxVanesaIglesias10
 

Recently uploaded (20)

Narcotic and Non Narcotic Analgesic..pdf
Narcotic and Non Narcotic Analgesic..pdfNarcotic and Non Narcotic Analgesic..pdf
Narcotic and Non Narcotic Analgesic..pdf
 
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...
Blowin' in the Wind of Caste_ Bob Dylan's Song as a Catalyst for Social Justi...
 
Faculty Profile prashantha K EEE dept Sri Sairam college of Engineering
Faculty Profile prashantha K EEE dept Sri Sairam college of EngineeringFaculty Profile prashantha K EEE dept Sri Sairam college of Engineering
Faculty Profile prashantha K EEE dept Sri Sairam college of Engineering
 
ICS2208 Lecture6 Notes for SL spaces.pdf
ICS2208 Lecture6 Notes for SL spaces.pdfICS2208 Lecture6 Notes for SL spaces.pdf
ICS2208 Lecture6 Notes for SL spaces.pdf
 
week 1 cookery 8 fourth - quarter .pptx
week 1 cookery 8  fourth  -  quarter .pptxweek 1 cookery 8  fourth  -  quarter .pptx
week 1 cookery 8 fourth - quarter .pptx
 
4.11.24 Mass Incarceration and the New Jim Crow.pptx
4.11.24 Mass Incarceration and the New Jim Crow.pptx4.11.24 Mass Incarceration and the New Jim Crow.pptx
4.11.24 Mass Incarceration and the New Jim Crow.pptx
 
How to Make a Duplicate of Your Odoo 17 Database
How to Make a Duplicate of Your Odoo 17 DatabaseHow to Make a Duplicate of Your Odoo 17 Database
How to Make a Duplicate of Your Odoo 17 Database
 
Unraveling Hypertext_ Analyzing Postmodern Elements in Literature.pptx
Unraveling Hypertext_ Analyzing  Postmodern Elements in  Literature.pptxUnraveling Hypertext_ Analyzing  Postmodern Elements in  Literature.pptx
Unraveling Hypertext_ Analyzing Postmodern Elements in Literature.pptx
 
Active Learning Strategies (in short ALS).pdf
Active Learning Strategies (in short ALS).pdfActive Learning Strategies (in short ALS).pdf
Active Learning Strategies (in short ALS).pdf
 
Mattingly "AI & Prompt Design: Large Language Models"
Mattingly "AI & Prompt Design: Large Language Models"Mattingly "AI & Prompt Design: Large Language Models"
Mattingly "AI & Prompt Design: Large Language Models"
 
Multi Domain Alias In the Odoo 17 ERP Module
Multi Domain Alias In the Odoo 17 ERP ModuleMulti Domain Alias In the Odoo 17 ERP Module
Multi Domain Alias In the Odoo 17 ERP Module
 
4.16.24 21st Century Movements for Black Lives.pptx
4.16.24 21st Century Movements for Black Lives.pptx4.16.24 21st Century Movements for Black Lives.pptx
4.16.24 21st Century Movements for Black Lives.pptx
 
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptx
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptxBIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptx
BIOCHEMISTRY-CARBOHYDRATE METABOLISM CHAPTER 2.pptx
 
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITW
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITWQ-Factor HISPOL Quiz-6th April 2024, Quiz Club NITW
Q-Factor HISPOL Quiz-6th April 2024, Quiz Club NITW
 
MS4 level being good citizen -imperative- (1) (1).pdf
MS4 level   being good citizen -imperative- (1) (1).pdfMS4 level   being good citizen -imperative- (1) (1).pdf
MS4 level being good citizen -imperative- (1) (1).pdf
 
Scientific Writing :Research Discourse
Scientific  Writing :Research  DiscourseScientific  Writing :Research  Discourse
Scientific Writing :Research Discourse
 
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptx
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptxINTRODUCTION TO CATHOLIC CHRISTOLOGY.pptx
INTRODUCTION TO CATHOLIC CHRISTOLOGY.pptx
 
Transaction Management in Database Management System
Transaction Management in Database Management SystemTransaction Management in Database Management System
Transaction Management in Database Management System
 
INCLUSIVE EDUCATION PRACTICES FOR TEACHERS AND TRAINERS.pptx
INCLUSIVE EDUCATION PRACTICES FOR TEACHERS AND TRAINERS.pptxINCLUSIVE EDUCATION PRACTICES FOR TEACHERS AND TRAINERS.pptx
INCLUSIVE EDUCATION PRACTICES FOR TEACHERS AND TRAINERS.pptx
 
ROLES IN A STAGE PRODUCTION in arts.pptx
ROLES IN A STAGE PRODUCTION in arts.pptxROLES IN A STAGE PRODUCTION in arts.pptx
ROLES IN A STAGE PRODUCTION in arts.pptx
 

3.1 A-Terpenoids-Menthol-Citral-Artemisin.pptx

  • 1. B.PHARM. 5th SEMESTER PHARMACOGNOSY ANDPHYTOCHEMISTRY-II BP504T. UNIT-III Isolation, Identification and Analysis of Phytoconstituents Terpenoids: Menthol, Citral, Artemisin
  • 2. What is terpenoids Any of a large class of organic compounds including terpenes, diterpenes, and sesquiterpenes. They have unsaturated molecules composed of linked isoprene units, generally having the formula (C5H8) n. Isoprene unit are 5 carbon unit For having isoprene unit they are also called isoprenoids
  • 3.
  • 4. Physical properties of terpenoids Odorous Volatile at room temperature They represents essence or active constituents of plants hence called essential oil Lighter than water Have high refractive index Are optically active
  • 5. Chemical Properties of teprenoids Terpenoids are the hydrocarbones of plant origin of the general formula (C5H8)n as well as their oxygenated, hydrogenated & dehydrogenated derivatives Terpenoids are regarded as derivatives of polymers of isoprene, CH3H2C=C- CH=CH2 (i.e.C5H80) They consist of a complex mixture of terpenes or sesquiterpenes, alcohols, aldehydes, ketone, acid & esters. They are unsaturated compounds. They undergo addition reaction with hydrogen, halogen, halogen acids to form addition products. They undergo polymerization and dehydrogenation in the ring. On thermal decomposition, terpenoid gives isoprene as one of the product.
  • 6. Chemical Properties of teprenoids Terpinoids are modified terpenes, where in methyl groups have been moved or removed, or oxygen atoms added. Sometimes terpinoids are added to proteins, e.g., to enhance their attachment to the cell membrane. The steroids and sterols in animals are biologically produced from terpinoid precursors.
  • 7.
  • 9. Menthol • Biological source: Menthol a monoterpene alcohol obtained from oils of Mentha piperita var. Vulgaris (Black peppermint) & Mentha piperita var. officinalis (White peppermint), • Family: Labiatae. Mentha species contains about 1-3% of volatile oils & oil contains not less than 44%. G.S.: Asia, Europe, North America, Cultivated in India, japan, Germany, France, Italy. Menthol is isolated by 2 methods Isolation Method 1: Hydro- distilation Method 2: Steam distillation
  • 10. Hydro-distillation Required quantity of coarse powder leaves of Mentha piperita. The peppermint oil is extracted by hydrodistillation method by using clevenger apparatus. The oil is separated from water and allow to cooling. After cooling crystals of (-) menthol will separate out. The crystals are collected by centrifugation and re- crystallized the menthol by acetone or any other low boiling point solvent.
  • 11. 1. Hydro-distillation by Clevenger apparatus system
  • 12. 2. Steam distillation Required quantity of air dried mentha plant is charged into stainless steel still having perforated bottom. The steam under pressure is generated with the help boiler and steam is passed through the drug. The mixture vapour (water and volatile oil) are passed through the condenser where vapour is cooled and back to liquid form. The mentha oil is collected in separating can. Mentha oil is floated on top of the water due to lighter than water. The oil is then decanted and filtered.
  • 14. 3. Method of Isolation Pippermint volatile oil- separated by steam & water distillation. To make oil free of moisture- it is passed through a bed of Na2SO4 The moisture free oil is frozen to -60°C for 7days by packing in tightly sealed plastic container. Menthol in oil separates as flaky crystals, separated by filtration. Mother liq. Containing some menthol along with other terpenes. The mother liq is treated with boric acid & further boiled for 3hr, followed by distillation for removal of menthone. Borate of menthol subjected to saponification with 50ml of NaOH by heating reflux for 1hr Resultant sol. Is allowed to cool for separation of remaining menthol crystals and mixed, dried in dessicator.
  • 15. Properties: • Appearance: White crystalline substances, which is solid at room temperature and melts slightly above (m.p. 41 to 43°C). • Odor: Characteristic and pleasant • Taste: Pungent followed by cooling sensation • Solubility: Soluble in 70% alcohol, ether and chloroform, insoluble in water • Identification by chemical test: • Few drops of sample is mixed with 5ml of nitric acid and heated on water bath. Blue color is developed within 5 minutes, after some time it becomes yellow which indicate the presence of menthol. • Small qty of menthol in TT + equal qty of thymol or camphor → liquifaction of contents in TT→ presence of Menthol • Crystals of Menthol in watch glass+ heat on water bath → material evaporates
  • 16. Analysis Take 10gm drug, add 10ml of acetic anhydride & 2gm of sodium acetate to the flask. Reflux for 1 hr. After cooling add 30ml water, boil it for 15 min. with continuous stirring. Then transfer in separating funnel. The oil layer is washed with water until it become neutral. Add 2 gm of sodium sulphate & shake it. Allow it for 30 min. & filter it. Take both layer in different flask, add 5 ml ethanol and Phenolphthelin indicator by adding 25 ml KOH ethanol solution. (for neutralize it) Again reflux for 1 hr. cool it titrated with HCl.
  • 17. • % Menthol = 7.814 x (c-a) {0.021 x (c-b)/U} A– 0.021 x (c-a) • Where, “a” & “b” = ml of HCl for oil “c” = ml of HCl for blank “U”= Un-acetylated oil “A”=Acetylated oil
  • 18. Analysis by TLC • Sample preparation: 1mg of Menthol is dissolved 1ml of methanol • Standard sample: Menthol • Stationary phase: Silica gel 60 F 254 • Mobile phase: Pure Chloroform OR hexane: EtOAc :: 8: 2 • Detecting agent: 1% vanillin-sulphuric acid reagent and heat the plate at 110°C for 10 minutes Anisaldehyde-sulphuric acid reagent, heat at 115°C-10min. • RF Value: 0.34, 0.48-0.62 Utilization • It is used as in various dosage forms for its cooling sensation, flavoring property, • carminative, antispasmodic and antipruritic Storage condition • It should be store in well closed and air-tight containers protected from light and in cool place.
  • 20. Citral Biological source: Citral is a monoterpene aldehyde obtained from oils of lemon grass Cymbopogon flexuosus and C. martini. Family: Graminae. It also present in Lemon oil (Citrus limonus) and Orange oil (Citrus aurantium). Family: Rutaceae. Lemon oil: Lemony, sweet smell, yellow in color. It contains not less than 75% of aldehyde calculated as Citral. Citral is isolated by following methods Isolation Method 1: Hydro- distillation Method 2: Fractional crystallization Method
  • 21. Hydro-distillation Required quantity of coarse powder leaves of lemon grass or other powder. The lemon grass oil is extracted by hydro distillation method by using Clevenger apparatus. The oil is separated from water and allow to cooling. After cooling of lemon grass will separate out.
  • 22. Citral The fresh plant material is hydro-distilled to obtain lemon grass oil. It is purified by fractional crystallization. To the total oil, first Sodium sulphite is added, the citrals get converted into its sulphite salt. The salt crystallizes out of the solution. The crystals are filtered and washed with ether or chloroform.
  • 23. Properties: • Appearance: Clear pale yellow liquid • Odor: Strong lemon like odour • Taste: Lemon like taste • Solubility: Soluble in 3 parts of 70% alcohol, chloroform and fixed oil. Insoluble in water. • Boiling point: 224-228 0C • Identification by chemical test: • Alcoholic solution of Sudan red III is added to the sample. Red colour is appeared which indicate presence of citral. • Tincture alkane is added to the sample. Red colour is appeared which indicate presence of citral.
  • 24. • Analysis by TLC • Sample preparation : 1mg of Citral is dissolved 1ml of methanol • Standard sample : Citral • Stationary phase : Silica gel –G • Mobile phase : Pure Chloroform • Detecting agent : 2, 4, dinitrophenyl hydrazine reagent • Color spots : Yellow to orange • RF Value : 0.51 • Utilization: • It is used as a flavouring agent and perfumery. Commercially citral is act as precursor for the synthesis of ß-ionone. ß-ionone is used as starting material for the synthesis of Vitamin A • Storage condition: • It should be store in well closed and air-tight containers protected from light and in cool place.
  • 26. Artemisin Biological source: Artemisin is a sesquiterpenoid lactone, obtained from the unexpanded flower- heads of Artemisia cina Berg, Artemisia brevifolia Wall, Artemisia maritime Linn. and other species Family: Asteraceae. It contains 2-3% of essential oil and two crystalline substances i.e Santonin and Artemisin. G.S: It is found in Italy, Iran, South Austrelia, In India Gujarat, J&K, Karnataka, UP and Hariyana
  • 27. Extraction Extraction can be done by 2 ways;- Soxhlet extraction Microwave-assisted extraction REQUIREMENTS • Raw material • Solvents (methanol, n-hexane, n-heptane, benzene, toluene, chloroform, methylene dichloride, ethyl acetate, acetone, acetonitrile..)
  • 28. SOXHLET EXTRACTION AND TEST SAMPLE PREPARATION 100 mg of fine powder was placed into an extraction thimble extracted with 170 ml of solvent via hot soxhlet extraction method for 6 hours over a water bath The extract was evaporated and redissolved in 5 ml methanol 10 μl of these test solutions was used for quantification purpose.
  • 29. MICROWAVE-ASSISTED EXTRACTION AND TEST SAMPLE PREPARATION 10 μl of these test solutions was used for quantification purpose. The extract thus obtained was evaporated and redissolved in 5 ml methanol For microwave-assisted extraction (MAE) the same parameters should be followed for every solvent Extraction parameters (160 watts, 120s, 10 ml per extraction cycle, two extraction cycles, and cleanup with 2 ml of corresponding solvent at the end of second cycle of extraction) 100 mg of fine powder was extracted under the influence of microwave energy using solvents
  • 30.
  • 31. Partition the extract using 50 ml hexane Partitioning done many times until hexane layer become colorless Two layers got from this process The methanol extract obtained was added 10 ml of distilled water Partitioned again using 50 ml of ethyl acetate Partitioning done many times until the ethyl acetate layer is colourless. Again we get 2 layers PARTITION OF EXTRACT 1. hexane extracts (non- polar fraction) 2. methanol extracts Each extract was concentrated using a rotavapor at a temperature of 40˚c. 2.methanol-water extract (polar fraction) 1.ethyl acetate extract (semipolar fraction)
  • 32. ESTIMATIONS By 2 methods:- • TLC Densitometer • HPTLC method TLC METHOD • Mobile phase :ethyl Acetate: hexane (3:97) • Stationary phase :60 F254 silica gel • Detecting reagent :anisaldehyde-sulphuric acid reagent • Spot volume :10 μL of test and standard sample spots • Spot colour :Pink-colored spots of artemisinin • Room temp. :25 +- 2˚C • Relative humidity :45+- 2%
  • 33. Extrapolating the graph to x axis give concentration of test sample Values are plotted in calibration curve (con Vs spot area) A simple TLC-densitometric technique has been developed for the rapid and accurate analysis of artemisinin in a at 540-610 nm using tungsten lamp large number of Artemisia annua plantlets
  • 34. HPLC METHOD •EQUIPMENT MODEL: -- WATERS 501 •COLUMN: -- WATERS C18 •DETECTOR: -- UV 260 nm •MOBILE PHASE: -- phosphate buffer : methanol (6:4) – PH – 7.9 •FLOW RATE: -- 1ml/min Calibration curve was plotted with diff conc
  • 35. Description - - colour less needles /white crystalline powder, odourless Identification - - TLC/ HPLC with std Molecular formula - - C15H22O5 Molecular wt - - 282.3 Solubility - - partially insoluble in water very soluble in dichloromethane MP - - freely soluble methanol, ethanol 151-154 c LOSS ON DRYING - - NMT 0.5% STORAGE - - Tightly closed container SPECIFIC SHEET
  • 36. Identification by chemical test: Sample is boiled with 10ml of alcohol and filtered. Sodium hydroxide is added to filtrate and heated. Red colour is appeared in liquid . • Analysis by TLC • Sample preparation: 1mg of Artemisin is dissolved Chloroform • Standard sample: Artemisin • Stationary phase: Silica gel –G • Mobile phase: Petroleum ether-Ethyl acetate (1:2) • Detecting agent: p-dimethyl amino benzaldehyde and heat at 80° C to produce color • RF Value: Compare with standard Artemisin
  • 37. Artemisnin • Analysis by UV • 1mg of Artemisin is mixed with 10ml of methanol and analysed λ200-400nm. • Utilization: • It is used as antimalarial. • It acts both against chloroquine sensitive and resistant P. falciperum and P. vivax malarial parasites. • Storage condition: • It should be store in well closed and air-tight containers protected from light and in cool place.