The document discusses the industrial production, estimation, and utilization of various phytochemicals, including sennoside, forskolin, diosgenin, digoxin, atropine, vincristine, and vinblastine, detailing their extraction methods and applications. It highlights the importance of these compounds in medicine, particularly in cancer treatment, and provides specific extraction and estimation procedures for each phytochemical. Additionally, it references notable figures in the field of phytochemistry and their contributions to drug discovery.

1. Unit IV
Industrial Production, Estimation and Utilization of
the Phytochemicals
Dr. Amit Gangwal
Associate Professor
Shri Vile Parle Kelavani Mandal’s
Institute of Pharmacy, Dhule
1

2. INDUSTRIAL PRODUCTION, ESTIMATION AND UTILIZATION OF PHYTOCHEMICALS
O
O
OH
OH
OH
OH
HO
O
OH
OH
OH
Forskolin
1. Sennoside
2. Artemisinin
3. Diosgenin
4. Digoxin
5. Atropine
6. Podophyllotoxin
7. Caffeine
8. Taxol
9. Vincristine and Vinblastine
10. Forskolin
11. Caffeine

4. INDUSTRIAL PRODUCTION, ESTIMATION AND UTILIZATION OF PHYTOCHEMICALS

6. Taxol

8. Sennoside

10. 1. Sennoside: Isolation and Utilization of Sennoside
 Sennosides are obtained from the dried leaflets of Cassia senna (Cassia acutifolia Delile and Cassia
angustifolia Vahl (Fam: Leguminosae).
 Senna leaves are powdered to 20-40 mesh and loaded into vertical/continuous extractors.
 Acetone at ambient temperature is circulated through the material to remove adherent impurities of
pesticides, and other acetone soluble unwanted material of no therapeutic value.
 It is then made free of acetone and extracted with 70% V/V alcohol (ethyl or methyl) pre-adjusted to pH
3.9 with citric acid at temperature 45-50°C.

11. The extraction is continued till washing show a positive test for
anthraquinones glycosides (colour reaction or TLC).
After extraction, the marc is discarded. The extracted liquid is filtered
and transferred to a tank fitted with stirrer.
The pH is adjusted to 6.0-6.2 with lime water.
It is then concentrated to a paste of 65-70% total solids in a multiple
effect evaporator.
The paste is dried in rotary vacuum drier at temperature 50-55°C. The
flakes obtained are pulverized to a fine powder.
It is then sifted to 80 mesh and packed preferably by vacuum sealing.

12. Estimation of Sennoside
• Hot water is useful for the extraction of anthraquinone glycosides.
• Acidified the aqueous extract and treat with chloroform to make free the
aglycone which is largely present in the solution.
• First neutralize the solution and then centrifuge it afterwards add ferric
chloride to the solution.
• Reflux the solution and further acidified it to bring oxidation and
hydrolysis.
• The aglycone present should be extracted with ether and re dispersed in
the magnesium acetate solution.
• Measure the absorbance of the sennoside and express the
concentration in comparison to sennoside B at 515 nm.

13. Forskolin
The botanical source of the crude drug is root of Coleus forskohlii synonym
Plectranthus barbatus belonging to Family Labiatae. The marker constituent of the
drug is forskolin (labdone diterpenoids).

14. 1. Roots of C. forskohlii are collected, washed, dried, and pulverised into granules.
2. The whole forskolin is extracted (utilising methanol as a solvent) in crude form by the
traditional methods.
3. The obtained methanol extract is concentrated, and the resultant concentrate is added
with chloroform.
4. Equal volume of water is added to the separating funnel.
5. The resulting mixture is shaken thoroughly, then allowed to settle, after which the
chloroform layer is separated.
6. The mixture is treated with water for 2 -3 times and the chloroform layer is separated and
concentrated.
7. A precipitate of forskolin is obtained by using ice cold n-hexane.
8. Forskolin is obtained as a reddish brown to brown coloured powder.

16. 3. Diosgenin
It is obtained from the dried tubers of Dioscorea deltoidea Wallich family: Dioscoreaceae
Industrial Production:
a. The tubers of the plant are cut into small pieces and dried under sun.
b. The dried tubers are powdered, extracted with ethanol or methanol twice for 6-8 hours.
c. It is filtered and the filtrate is concentrated to a syrupy liquid.
d. The concentrated liquid is then hydrolysed using an acid, hydrochloric acid or sulphuric acid for
2-12 hrs.
e. About 85 % of the crude Diosgenin is precipitated.
f. The precipitate are filtered, washed with water and purified by alcohol.

17. Estimation of Diosgenin
Thin Layer Chromatography of Diosgenin:
 Sample Preparation: Dissolve Diosgenin in methanol (1:1 w/v)
 Stationary Phase: Silica gel plates
 Mobile Phase: Toluene: Ethyl acetate (7:3)
 Rf: 0.37

18. By UV standard curve method
• Prepare the solution A (0.5 ml ρ-anisaldehyde in 99.5 ml ethyl acetate) and
solution B (50 ml sulphuric acid with 50 ml ethyl acetate).
• The test samples is dissolved in 2 ml ethyl acetate and add 1 ml of reagent A and
B.
• Stirred well and maintain the temperature 60° C for 10 minutes to develop the
colour.
• Allow to cool at 25°C and measure the absorbance at 430 nm using ethyl acetate
as blank.
• Similarly the calibration curve of standard diosgenin (2-70 μg) in ethyl acetate
was made and determine the concentration of unknown sample.

19. Pharmacognosy & Phytochemistry-II
4. Digoxin
 Digoxin or lanoxin is the most widely used cardiac glycoside obtained from Digitlais lanata family:
Scrophulariaceae.
 It is a secondary glycoside which is produced from a primary glycoside i.e Lanotoside C.
 Its hydrolysis yields three molecules of Digitoxose sugar (glycone) and digoxigenin (aglycone).
 It is highly potent drug and should be handled with exceptional care.
INDUSTRIAL PRODUCTION, ESTIMATION AND UTILIZATION OF PHYTOCHEMICALS

20. • The dried powdered leaf extracted with petroleum ether.
• Discard the extract and digested the marc with water at 0-4°C so that the
polysaccharide may remove. Again the filtrate is discarded and the marc is extracted
with alcohol and water.
• Under reduced pressure alcoholic extract is concentrated at 50°C. This concentrate
solution is treated with lead acetate to remove the impurities. In the water soluble
glycoside portion (obtain after the treatment of lead acetate) maintain the pH around 6
and then wash with non polar solvent.
• Discard the organic layer and treat the aqueous layer with 0.5 percent sodium sulphate
to remove the lead sulphate. The aqueous layer is then treated with chloroform and
afterwards with ethanol.
• Chloroform portion contain less polar glycoside while ethanolic portion contain more
polar glycoside. The ethanolic portion further treated and subjected to chromatography
to separate the digoxin.

22. • Atropine is an alkaloid isolated from the Hyoscyamus niger (leaf
and flowering tops) or dried leaves of Hyoscyamus muticus or
Atropa belladonna (aerial parts) or Atropa acuminata family
Solanaceae.
• Atropine is a poisonous product and its production on synthetic
way is costly compare to extraction from natural way.
• Atropine is needle like crystal, white colour or colourless, optically
inactive usually present with laevorotary hyoscyamine.
• The melting point of atropine is 115-116°C.
Atropine

23. Atropine
Method of Industrial Production:
a. The powdered drug material is thoroughly moistened with an aqueous solution of
sodium carbonate and then extracted with ether or benzene.
b. The alkaloidal free bases are extracted from the solvent with water acidified with acetic
acid.
c. The acid solution is then shaken with solvent ether to remove colouring matter.
d. The alkaloids precipitated with sodium carbonate, filtered off, washed and dried.
e. The dried mass is dissolved in solvent ether or acetone and dehydrated with anhydrous
sodium sulphate before filtration.
f. The filtrate after concentration and cooling yields crude crystals of hyoscyaine and
atropine from the solution.
g. The crude crystalline mass obtained after filtration is dissolved in alcohol, and sodium
hydroxide solution is add the mixture is allowed to stand until hyoscyamine is
completely racemized to atropine which indicated y the absence of optical activity.

24.  The crude atropine is purified by crystallization from acetone.
 Atropine sulphate is the most important salt of atropine.
 It occurs in form of colourless crystalline powder. It is soluble in water and alcohol but insoluble in
ether and chloroform.
Melting Point: 115-116⁰C
Estimation of Atropine: TLC Profile
 Sample Preparation: Dissolve 1% of atropine in 2N acetic acid is spotted over silica gel G plate and eluted in
the solvent system of strong ammonia solution: methanol (1.5:100).
 The TLC plate is spread with an acidified iodoplatinate solution.
 Atropine gives the Rf value 0.18.

25. Estimation
• Add alcohol in atropine sulphate and evaporate the alcohol.
• Dissolve the residue again in alcohol and add calculated quantity of 0.1N HCl in
this alcohol and titrate excess of this acid with 0.1N NaOH using methyl red as
indicator.
• Each ml of 0.1 N HCl is equivalent to 0.3384 gm of atropine sulphate.
• In another method atropine sulphate (around 0.1 gm) was weighed and
dissolved in glacial acetic acid (50 ml).
• Titrate this solution with 0.1N per chloric acid and determine end point by
potentiometrically.
• Make the necessary correction by performing the blank determination.
• Factor 1 ml of 0.1N HClO4 ≡ 0.0670 gm of atropine sulphate.

26. 1. Atropine sulphate acts as anticholinergic drug.
2. It is used in surgery as antisialogogue to reduce nasal, salivary,
bronchial and pharyngeal secretion.
3. It is dispensed through intra muscularly injection before the
anesthesia.
4. Atropine is used as an antidote for the poisoning of
organophosphate insecticide and physostigmine.
5. It also shows mydriatic and anti spasmodic property. In
ophthalmology it is also used to measure the refractive error.
Uses

27. Vincristine and Vinblastine
Both these indole alkaloids are isolated from the dried entire plant of Catharanthus roseus Linn (Apocynaceae)

28. 28
Although the plant has a certain reputation in
folk medicine for the treatment of diabetes,
modern investigators have been unable to
confirm this property.
Instead Canadian workers, during 1955–1960,
discovered that extracts of the leaves
produced leukopenic actions in rats.
These observations led researchers at Eli Lilly
to undertake an intensive phytochemical
investigation of the plant with a view to the
isolation of constituents of value in cancer
chemotherapy.
Six alkaloids proved active in this respect and
two are now available commercially

29.  Industrial Production of Vinblastine:
a) The dried leaf material is taken and is extracted with a solution of hot ethanol-water-
acetic acid in a ratio of 9:1:1.
b) The solvent is removed and to the residue hot 2 % hydrochloric acid solution is added.
c) The pH of the acidic extract is adjusted to 4, for the precipitation of the non-alkaloidal
components, which can be separated by filtration.
d) The pH of the aqueous acidic solution is now adjusted to 7 and then extracted with
benzene.
e) The benzene layer is evaporated to obtain vinblastine and other alkaloids.

31. • Because these alkaloids are only minor constituents of the plant
(vincristine is obtained in about 0.0002% yield from the crude drug),
large quantities of raw material are required and chromatographic
fractionations are extensively employed in the isolation procedures.
• In addition, there is a growing demand for vincristine rather than
vinblastine, but the plant produces a much higher proportion of
vinblastine.
• Fortunately, it is now possible to convert vinblastine into vincristine
either chemically, or via a microbiological N-demethylation using
Streptomyces albogriseolus.

32. As such, access to these chemicals is only possible through extraction from the Madagascan
periwinkle.
Methods have been developed to synthesize these drugs starting from precursors, called
catharanthine and vindoline, also isolated from the plant.
However, today approximately 500 Kg of dried leaves are required to produce 1 g of vinblastine.
In this context, the possibility to increase the yield through synthetic biology would allow to
overcome the challenges of having a steady supply and possibly decrease the price of the
drugs, allowing more people to have access to treatment.

33. Isolation of Vinblastine and Vincristine
i. The phenolic materials are removed by the washing the extract with dilute alkali.
ii. The washed extract is subjected to chromatography on alumina and elution is
carried out in 18 fractions starting with benzene-methylene chloride (65:35) mixture
to pure methylene chloride.
iii. Vinblastine recovered in the ninth fraction. Further elution of the column results in
separating the fractions of vincristine.
Estimation of Vinca alkaloids
Thin Layer Chromatography of Vincristine: Vincristine dissolved in 25% water in
methanol solution, spotted in silica gel-G plate and developed using the solvent,
acetonitrile:benzene (30:70). The dried plates are sprayed with 1% solution of ceric
ammonium sulphate in 85 % of phosphoric acid. The Rf value of the appeared spot
would be 0.39.

34. Vincristin and vinblastin sulphate are estimated with the help of HPLC. The following solution are
prepared for the estimation:
Solution 1: 0.1 percent w/v of the substance being examine.
Solution 2: Contain 0.2 percent w/v each of vinblastine sulphate RS and vincristine sulphate RS
solution.
Solution 3, 4 and 5: 0.1 percent w/v, 0.002 percent w/v and 0.0001 w/v respectively of vinblastin
sulphate (if vinblastin estimated) or vincristin sulphate (if vincristin sulphate estimated).
Column: Packed with stationary phase LC2.
Flow rate: 1.0 ml/min.
Mobile phase: (For vinblastin) Mixture of 70 volume of methanol 30 volume of 1.5 percent w/v of
diethylamine (pH adjusted 7.5 with phosphoric acid).
Detection wavelength: (For vinblastin sulphate) 297 nm.
Mobile phase: (For vincristin sulphate) Mixture of 50 volume of methanol 38 volume of 1.5 percent w/v
of diethylamine (pH adjusted 7.5 with phosphoric acid) 12 volume of acetonitrile.
Detection wavelength: (For vincristin sulphate) 262 nm.

35. • Vinblastine is used mainly for the treatment of generalized Hodgkin’s disease, and non-
Hodgkin’s lymphomas.
• Vincristine is used principally in the treatment of acute lymphocytic leukaemia in children. It
has other applications for lymphomas, small-cell lung cancer, cervical and breast cancers.
• The semi-synthetic vindesine is also used in the treatment of acute lymphoid leukaemia in
children.
• Vincristine has a superior antitumour activity compared to vinblastine, but is more neurotoxic.
• Vinorelbine is a newer, orally active, semi-synthetic anhydro derivative of 8′-norvinblastine
with a broader anticancer activity and lower neurotoxic side-effects than the other
Catharanthus alkaloids.
Utilisation of Vinca alkaloids

36. Taxol:Taxol is a diterpenoid alkaloid obtained from bark of plant Taxus brevifolia, of family
Taxaceae. It is a potent antineoplastic agent.
Monroe Eliot Wall and Mansukh Wani (Left)

37. • Professor Mansukh C. Wani, Ph.D. (died 2020), was a principal scientist (emeritus) at
the Research Triangle Institute in North Carolina.
• He was co-discoverer of Taxol and camptothecin, two anti-cancer drugs considered
standard in the treatment to fight ovarian, breast, lung and colon cancers.
• In 2000, Dr. Wani received an award for applied research in medicine, the Charles F.
Kettering Prize, from the General Motors Cancer Research Foundation.
• Dr. Wani was born in Nandurbar, Maharashtra, India.
• He attended the University of Bombay, receiving a bachelor's degree in 1947 (in
chemistry) and a master's in 1950 (in organic chemistry).
• He moved to the United States in 1958, and received his PhD from Indiana University
Bloomington in 1962, when he joined Research Triangle Institute.
• He used to live in Durham, North Carolina. He died on 11 April 2020.

38. • The research team of Dr. Monroe E. Wall and Dr. Mansukh C. Wani of
Research Triangle Institute discovered two first-in-class life-saving
chemotherapeutic agents.
• Camptothecin, first isolated and identified from Camptotheca acuminata,
was found to kill cancer cells uniquely via topoisomerase I poisoning.
• Presently, two first-generation analogues of camptothecin are used to treat
ovarian, colorectal, and small-cell lung cancers, and several second-
generation analogues are in clinical trials.
• Taxol, first isolated and identified by Wall and Wani from Taxus brevifolia,
was found to inhibit cancer cell growth via the stabilization of microtubules.
• In 1992, taxol was approved for refractory ovarian cancer and today is used
against breast and non-small cell lung cancers.

39.  Industrial Production of Taxol:
i. Take the accurately weighed quantity of bark powder of Taxus plant. Extract the powder exhaustively with ethanol or
methanol.
ii. Filter and evaporate filtrate at 40 ⁰C to dryness. Dissolve this residue again in methanol.
iii. Filter and evaporate the filtrate to dryness. Now dissolve this residue in a mixture of carbon tetrachloride and water.
iv. Filter and centrifuge. Separate and combine carbon tetrachloride layer. Evaporate this layer to dryness.
v. Dissolve this residue in mixture of methanol and carbon tetrachloride (1:1). Filter and evaporate to obtain dry residue of
taxol alkaloids.

40.  This crude residue can be further purified by preparative TLC using silica gel as stationary phase and mobile phase
of carbon tetrachloride and methanol (95:5). Rf value: 0.35.
Utilization of Taxol
 As a potent anticancer drug. Used in treatment of ovarian, lung, bladder, esophageal & other types of cancers.
 Paclitaxel Injection is a clear, colorless to slightly yellow viscous solution. It is supplied as a non-aqueous solution
intended for dilution with a suitable parenteral fluid prior to intravenous infusion. Paclitaxel is available in 30 mg (5
mL), 100 mg (16.7 mL), and 300 mg (50 mL) multidose vials.

42. Taxol is an especially versatile drug. It is used for breast cancer in the early
stages as well as for metastatic breast cancer.
Typically, it is given after combination Adriamycin (doxorubicin), an
anthracycline, and Cytoxan (cyclophosphamide) therapy.
It is also used prior to surgery in cases when it is necessary to shrink a
tumor before it's removed (neoadjuvant chemotherapy).
Besides breast cancer and ovarian cancer, Taxol can be used to treat several
other types of cancer, including lung cancer and Kaposi's sarcoma, a rare type of
skin cancer that affects people with AIDS.
The chemotherapy class taxanes includes the drug Taxotere (docetaxel) as well
as Taxol.

44. 9. Podophyllotoxin
Indian podophyllum is the root and rhizome of Podophyllum hexandrum Royle (Berberidaceae)
 Industrial production of Podophyllotoxin:
a) Extract the powdered rhizome/roots of P. emodi with methanol.
b) Then it is reduced under vacuum. Semisolid mass is put into acidulated water (10 ml HCL in 100
ml water).
c) The precipitates are allowed to settle. Filtrate is decanted and then washed with cold water.
d) Resin obtained is dried and upon drying, it gives dark brown amorphous powder called
podophyllin.
 Another method of extraction: Pure podophyllotxin is obtained by dissolving the chloroform
soluble fraction in alcohol. Then it is refluxed with neutral aluminium oxide so that solution
becomes light yellow. To alcoholic solution benzene is added which yielded Podophyllotoxin of
95-98% purity.

45.  Estimation of Podophyllotoxin: TLC profile of Podophyllotoxin:
 Preparation of sample: 1 mg dissolved in methanol
 Stationary phase: Silica gel GF
 Mobile phase: Toluene: ethyl acetate (5:7)
 Detecting agent: Sulphuric acid
 Rf: 0.39

46. Method 2: HPLC Determination of Podophyllotoxin Column
Mobile Phase
Methanol:water (62:38).
Flow Rate
1.0ml/min.
UV detection at 280 mm.
Standard Preparation
Solution of p odophyllotoxin (of known concentration) is prepared in methanol within the linearity range (20 g–2 g).
Sample Preparation
1) The powdered sample (2gm) is refluxed with 50ml methanol for an hour, cooled, and filtered.
2) The marc is refluxed for another 1 hour with 50ml methanol.
3) The filtrates of both the cycles of refluxes are combined and evaporated to dryness under vacuum.
4) The residue is dissolved in 20ml methanol and volume is made up to 50ml with the same solvent.
Procedure
5ml each of standard and sample preparations are injected to HPLC on triplicate.
Then the peak areas for podophyllotoxin are recorded, and the yield % in the sample is calculated.

47. 10. Caffeine
Source: xanthine alkaloid obtained from the leaves of Camellia sinesis (Theaceae), seeds of
Coffea arabica (Rubiaceae).
The most commonly known sources
of caffeine are coffee, cocoa beans,
cola nuts and tea leaves. Caffeine is a
bitter, white crystalline xanthine
alkaloid that is a psychoactive
stimulant drug. Caffeine is one of the
most naturally occurring methyl
derivatives of xanthine. Black tea
contains caffeine (0.5%). Theobromine
(0.017%) and theophylline (0.13%).

49. • Teal leaves’ powder is boiled with 2% sodium carbonate for 10 min & filtered in hot
condition.
• Evaporate & partitioned with dichloromethane.
• Evaporate to get crystals of caffeine.
• Purified by recrystallization from hot ethanol.
Industrial production of caffeine

50. 1. Place 15 g of tea leaves, 5 g of calcium carbonate powder and 200 mL of water
into a 600 mL beaker.
2. Boil the solution on a hot plate for 20 minutes with occasional stirring.
3. Cool the solution but, while it is still warm, vacuum filter through a Buchner
funnel using a fast filter paper, if available.
4. Rinse the leaves with 50 mL of water. Carefully press out as much filtrate as
possible since the caffeine is in the aqueous layer. Rinse again with 50 mL of
water.
5. Cool the solution to room temperature and pour it into a 500 mL separatory
funnel. Extract with 35 mL of methylene chloride. Then shake vigorously for 10
seconds and relieve pressure, repeat the shaking two more times. An emulsion
will probably form.

51. 6. To break the emulsion formed in the methylene chloride layer, slowly
drain the methylene chloride layer through a small amount of anhydrous
magnesium sulphate in a powder funnel with a loose cotton plug (a tight
plug will prevent drainage).
7. Extract the aqueous solution once again with a 35 mL of methylene
chloride, repeating the steps above to collect the lower layer. Combine
the methylene chloride extracts and, if necessary, dry further with
additional anhydrous magnesium sulphate.

54. Estimation through HPLC method
Assay: Weigh accurately about 0.4g of caffeine and dissolve it in 40ml of acetic anhydride cool and add
80ml of benzene and titrate with 0.1N perchloric acid determining the end point by potentiometrically by
performing a blank titration and make necessary correction. Each ml of 0.1N perchloric acid is
equivalent to 0.01942g of caffeine.

56. Utilization
• Stimulant and diuretic
• Increases the blood pressure.
• Stimulates the central nervous system.
• Promotes urine formation.
• Stimulates the action of the heart and
lungs.
• Treats migraine.
• Increases the potency of analgesics.
• Relieves asthma attacks.

57. 11. Artemisinin
Source : It is a sesquiterpene lactone
obtained from the leaves and the
closed unexpanded flower heads of
Artemisia annua, belonging to the
family Asteraceae.

58. Malaria is a major cause of human morbidity and mortality in the tropical endemic countries
worldwide. This is largely due to the emergence and spread of resistance to most antimalarial
drugs currently available.
Based on the World Health Organization recommendation, artemisinin-based combination
therapies are now used as first-line treatment for Plasmodium falciparum malaria.
Artemisinin or qinghaosu (Chinese name) and its derivatives are highly potent, rapidly acting
antimalarial drugs.
Artemisinin was discovered in 1971 by a Chinese medical scientist Youyou Tu, who was
awarded the Nobel Prize in 2015 on her discovering the antimalarial properties of qinghaosu
from the traditional Chinese qinghao plant.
Nevertheless, artemisinin resistance in falciparum malaria patients has first emerged on the Thai-
Cambodian border in 2009, which is now prevalent across mainland Southeast Asia from Vietnam
to Myanmar.

61. Tu Youyou, born 30 December 1930, is a Chinese pharmaceutical chemist and malariologist.
She discovered artemisinin and dihydroartemisinin, used to treat malaria, a breakthrough in twentieth-
century tropical medicine, saving millions of lives in South China, Southeast Asia, Africa, and South
America.
For her work, Tu received the 2011 Lasker Award in clinical medicine and the 2015 Nobel Prize in
Physiology or Medicine jointly with William C. Campbell and Satoshi Ōmura.
Tu is the first Chinese Nobel laureate in physiology or medicine and the first female citizen of
the People's Republic of China to receive a Nobel Prize in any category.
She is also the first Chinese person to receive the Lasker Award. Tu was born, educated and carried out
her research exclusively in China.

62. In 1967, a drug discovery project was set up in China, named Project 523, conducted by Youyou Tu at the Chinese
Academy of Medical Sciences in Beijing.
Her team had screened over 2000 traditional Chinese recipes and made 380 herbal extracts which were tested on
malaria-infected mice.
A herbal extract used for over 1600 years in traditional Chinese therapy for “intermittent fever” the hallmark symptom
of malaria, was found effective.
The extract from qinghao or huanghuahao (Artemisia annua L.), named qinghaosu, was isolated by low temperature
ethyl ether extraction and chemically characterized in 1971.
The active antimalarial moieties and the physicochemical properties were determined in vitro and in vivo in both
animal models and in human.
The drug was distributed to the rest of the world in 1979. Tu was awarded the Nobel Prize in Medicine on October 5,
2015, for her discovery of qinghaosu/artemisinin and the more potent derivative dihydroartemisinin, effective
antimalarial drugs which saved millions of lives.

63. Industrial production
• Fresh leaves are dried below 60˚C, powder is extracted with methanol
by maceration.
• Methanol extract partitioned with hexane to remove unwanted material.
Mixed with water and hydro-alcoholic extract is obtained.
• The hydro alcoholic extract partitioned with ethyl acetate until the
colorless and concentrated at controlled temperature at 40˚C under
vacuum.
• Artemisinin obtained as fine white crystals after recrystallization with
cyclohexane.

65. Artemisinin and its derivatives are endoperoxide-
containing natural products that are being used widely as
antimalarials in many parts of the world.
The most commonly used artemisinin derivatives
are artesunate, artemether, arteether, and
dihydroartemisinin.

67. Estimation
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-coloured spots of artemisinin
• These compounds lack physicochemical properties; therefore, detection by
using direct UV does not provide the required sensitivity and selectivity for
the analysis of trace amounts of these compounds.
• Chromatographic column treated with alkalis is used to detect a stable
product of artemisinin with maximum absorption at 289nm. It is used for
metabolites having peroxide linkages.
• The sensitivity of various detection methods can be enhanced by
chromogenic derivatisation methods.

68. Utilization
• Mainly as antimalarial
• In gastric infections
• Suppress inflammatory immune
reactions
Predecessor of
Artemisinin