corrected hand book - isca 978-93-84648-67-1.pdf · a hand book on: semi micro ... this handbook...
TRANSCRIPT
![Page 1: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/1.jpg)
By
Dr. G. Madhumitha & Ms. J. Fowsiya
International E – Publication
www.isca.me , www.isca.co.in
A HAND BOOK ON: SEMI MICRO
TECHNIQUE FOR EXTRACTION
OF ALKALOIDS
![Page 2: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/2.jpg)
A HAND BOOK ON: SEMI MICRO
TECHNIQUE FOR EXTRACTION
OF ALKALOIDS
By
Dr. G. Madhumitha
M.Sc., M. Phil., Ph.D., MBA (HR), F.I.S.C.A.
Assistant Professor,
Chemistry of Heterocycles and natural product research laboratory,
School of Advanced Sciences,
VIT University, Vellore 632014, Tamilnadu, India.
Email: [email protected], [email protected]
&
Ms. J. Fowsiya Chemistry of Heterocycles and natural product research laboratory,
Organic Chemistry Division,
School of Advanced Sciences,
VIT University, Vellore 632014, Tamilnadu, India.
2015
International E - Publication
www.isca.me , www.isca.co.in
![Page 3: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/3.jpg)
International E - Publication 427, Palhar Nagar, RAPTC, VIP-Road, Indore-452005 (MP) INDIA
Phone: +91-731-2616100, Mobile: +91-80570-83382
E-mail: [email protected] , Website: www.isca.me , www.isca.co.in
© Copyright Reserved
2015
All rights reserved. No part of this publication may be reproduced, stored, in a
retrieval system or transmitted, in any form or by any means, electronic,
mechanical, photocopying, reordering or otherwise, without the prior permission
of the publisher.
ISBN: 978-93-84648-67-1
![Page 4: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/4.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS iii
PREFACE
A Hand Book on: Semi Micro Technique for Extraction of Alkaloids published by
ISCA provides information on the alkaloids, its classification and the various methods
such as conventional and non-conventional methods involved in the extraction of
alkaloids from different sources. This hand book serves as a dictionary on extraction of
Alkaloids for under graduate and post graduate chemistry students. First, considerably
greater emphasis has been placed on extraction and identification of compounds from the
natural source by simple chemical tests. Secondly, whilst retaining undiminished the full
and clear directions provided for students who are studying the Phytochemistry/
Pharmacognosy/ Natural Product Chemistry. We have extended the scope of the work so
that it covers most of the needs of students working for an Honors/Special degree. These
experiments form the central building in pillar in Phytochemical approach around which
most of the recent research developments revolve. We have maintained the standard
which was self-imposed when this book was written, namely, that all the experiments in
the handbook had been critically examined.
DR. G. MADHUMITHA
&
Ms. J. FOWSIYA
![Page 5: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/5.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS iv
Index
S.no Content Pg.no
1 Introduction 1
2 General characteristics of alkaloids 1
3 Classification of alkaloids 2
4 Extraction of alkaloids 5
4.1 Different extraction methods of alkaloids 5
4.2 Sample preparation 6
4.3 Choice of solvent 6
4.4 Soxhlet extraction 7
4.5 Stas otto process (method-1) 9
4.6 Stas otto process (method-2) 10
4.7 Kippenberger’s process 11
4.8 Maceration 12
4.9 Manske’s process 13
4.10 Negative pressure cavitation extraction 14
4.11 Microwave assisted extraction 15
4.12 Accelerated solvent extraction 16
4.13 Subcritical water extraction 17
4.14 Ultrasonic assisted extraction 18
4.15 Pulse electric field extraction 19
4.16 Merck process 20
4.17 Thiboumery and Mohr process 21
4.18 Robertson-gregory process 22
5 Separation of different types of alkaloids 23
6 Qualitative analysis of alkaloids 24
![Page 6: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/6.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 1
1. Introduction
Phytochemistry deals with chemical constituents of plants, phytochemicals structure, and
their biosynthesis and biological activities. Phytochemicals referred as biological active
components of plants. The plant constituents were classified into primary and secondary
metabolites. The primary metabolites are sugar, amino acids, nucleic acid, etc. Whereas
secondary were alkaloids, anthocyanin, flavonoids, terpenoids, etc., [1]
. The quantities of these
metabolites vary from species to species [2]
. The Alkaloids were the most important group of
secondary metabolites found in living organism. Alkaloids never occur alone, they were a
mixture of few major and few minor alkaloids. It has different types of structure, properties and
pharmacological activities. Most of the alkaloids contain nitrogen in negative oxidation state and
also in addition to carbon, hydrogen it contains sulphur, oxygen and phosphorus.
In 1803 semi-pure alkaloid was isolated by Derosne and in 1805 Serturner isolated and
characterized the opium alkaloids from Papaver somniferum. The first synthesized alkaloid was
coniine from Conium maculatum in 1886. Strychnine, Emetine, Brucine, Piperine, Caffeine,
Quinine, Cinchonine and Colchicine alkaloids are the cornerstone of all that has occurred in
alkaloids chemistry to the present day [3]. Most of the alkaloids were derived from amine by
decarboxylation of amino acids.
The Alkaloids are biologically active compounds used for anesthetics, sedatives, stimulants,
Tranquilizers. Today, several alkaloids are still useful such as caffeine used for the mental
alertness, Cocaine used as anesthetic and Morphine used as the painkiller [4]
.
The present study is on the extraction of known and unknown alkaloids from varies plant
species with new techniques of modern science. This handbook provides an easy and brief
explanation for the different extraction methods and qualitative tests for alkaloids.
2. General characteristics of alkaloids
Most of the alkaloids are bitter taste, weak base, colorless, poorly soluble in water and
readily soluble in organic solvent such as diethyl ether, chloroform, etc., a few alkaloids are
colored such as Berberine is yellow color and salt of Sanguinarine is copper Red color. The
![Page 7: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/7.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 2
Alkaloids will be decomposed by heat except strychnine and caffeine. Mostly the physical form
of alkaloids is crystalline solids and few are amorphous solids [8]
.
3. Classification of alkaloids
When compared with other class of naturally occuring conpounds, there is no uniform structure
classification for alkaloids. Present days alkaloids classified based on the carbon skeleton that
present in the alkaloids [9]
.
Scheme 1. Types of Alkaloids
• Atrophine
• Nicotine
• Morphine
True alkaloids: It contains heterocyclic ring with Nitrogen and origenated from amino acids
• Ephedrine
• Mescaline
• Adrenaline
Proto alkaloids: No heterocyclic ring with Nitrogen and derived from amino acids
• Caffeine
• Theobromine
• Theophylline
Pseudo alkaloids: Contains heterocyclic ring with Nitrogen but not derived from amino acids
![Page 8: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/8.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 3
Apart from the above classification, the following methods can be employed in the classification
of alkaloids, they are as follows.
Scheme 2. Classification of alkaloids
In general, alkaloids classified into two division, i) Heterocyclic ii) Non-Heterocyclic alkaloids
The following are few examples for Heterocyclic and Non-heterocyclic alkaloids.
Fig 1. Types of Alkaloids based on their Structure
Biosynthetic classification
• Indole
• Piperidine
• Pyrrolidine
• Phenylethylamine
• Imidazole
Chemical classification
• Tropane
• Quinoline
• Purine
• Diterpene
• Steroidal
Pharmacological classification
• Morphine
• Quinine
• Lobeline
• Aconitine
• Ergonovine
Taxonomic classification
• Cannabinaceous
• Rubiaceous
• Solanaceous
![Page 9: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/9.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 4
Scheme 3: A few examples for Heterocyclic and Non-Heterocyclic alkaloids
4. Extraction of Alkaloids
The extraction is defined as the analytical tool for the separation of biologically active
components of plants or animals from inert or inactive component by using suitable solvent in
Standard extraction procedure. Maceration, percolation, digestion, soxhlet extraction,
microwave-assisted extraction (MWE), sonication (USE), super fluid extraction (SFE) is general
techniques for the extraction of alkaloids [10]
. The extract composition is depending on, extraction
Heterocyclic Alkaloids
Pyrrole
Pyrrolidine
Pyridine
Tropane
Quidine - Quinoline
Isoquinoline
Aporphine
Purine
Indole
Terpenoid
Non-Heterocyclic Alkaloids
N-Methyltramine
Ephedrine
Pachysandrine
Mescaline
Erythromycin
Colchicine
Jurubin
Taxol
![Page 10: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/10.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 5
method, temperature, solvent nature, concentration of solvent, polarity, solubility and extraction
time. These traditional methods have some limitation including, long time extraction, solvent
consumption, less efficiency etc., but in recent year new extraction techniques have been
developed with more efficiency, less consumption of solvent and easy to isolation
[11]. The
following extraction methods were outstanding development in the area of extraction of different
alkaloids, spectroscopic techniques, to determine activity of alkaloids and structural elucidation.
4.1. Extraction methods of alkaloids
The following methods were used for the extraction of alkaloids from various natural sources.
1. Soxhlet extraction.
2. Stas-otto extraction
3. Kippenberger’s process
4. Manke’s process
5. Microwave assisted extraction (MAE)
6. Ultrasonic assisted extraction (UAE)
7. Subcritical water extraction (SWE)
8. Maceration
9. Negative pressure cavitation extraction (NPCE)
10. Accelerated solvent extraction (ASE)
11. Heat reflux extraction
12. Pulse elecric field extraction (PEFE)
13. Merck process
14. Thiboumery and Mohr process
15. Robertson Gregory process
![Page 11: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/11.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 6
Scheme 4: Conventional, Non-conventional method
4.2. Sample preparation
The fresh plant material was used for the extraction and the source has been kept in
alcohol for few minutes. But most commonly dried material of the plant species were used for
the extraction. The drying method should be carried out at controlled condition without using
high temperature in air drier. This dried plant can be stored in a container for long time. The
plant material is reduced to a coarse powder using grinders and sieves to facilitate maximum
effective contact of the solvent with alkaloids and tissues.
4.3. Choice of solvent
The solvent act an important role in the extraction process and its choice depends on the
plant material. Commonly alcohol, ethyl acetate, chloroform and water were used as solvent to
obtain high yield. The alcohol used for the pretreatment is to remove chlorophyll and impurities.
The solvent should possess the following property such as low toxic, ease in handling and
storage, chemically inert. A good solvent must have the following properties,
• Soxhlet
• Maceration
• Stas Otto method
• Kippenberger's process
Conventional
• MWE
• SWE
• NPCE
• PEFE
Non-Conventional
![Page 12: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/12.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 7
Scheme 5: Good qualities of solvent
4.4. Soxhlet extraction
The main advantage of the Soxhlet extraction is, it is continuous
extraction process, less extraction time and sample gets continuous
contact with solvent, no filtration required and low cost. There is some
drawback of this method when compared with other techniques. The
drawbacks are, large amount of extractant wasted, sample should be
boiled over boiling point of solvent for long time which cause
decomposition of thermo-labile component and more solvent consuming
method [12]
• The powdered plant material taken in the fat free cellulose
extraction thimble and covered with cotton.
• A suitable solvent is added to the chamber.
• Soxhlet chamber, condenser and collecting flask
assembled in correct manner and heat the setup under reflux [13]
.
Goo
d q
uali
ties
of
solv
ent
Non-inflammable
Non-toxic or low toxic
Easily volatile
Low Eco-impact
In-expensive
High purity
Sholud give high yield of product
Low polarity
Chemically inert with other components
Easily Recoverable
![Page 13: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/13.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 8
Fig. 2: Soxhlet Extraction
Residue dried in oven at
103°C
Ground sample
Evaporate the solvent
Sample dried in
extraction thimble
Sample weighed in
extraction thimble
Moist sample dried for
2-3 h at 103 °C
Sample weighed in
extraction thimble
Liquid sample weighed
and evaporated in dish
Add sand and anhydrous
sodium sulfate
Close the sample with fat free cotton
wad and taken in soxhlet apparatus
Fill the solvent in solvent vessel at 110-130°C
for 20-30 cycles of extraction (4-6 h)
![Page 14: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/14.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 9
4.5. Stas otto process (Method-1)
Method 1: The solid ingredients were moistened with appropriate amount of solvent and
allowed to stand for 4 h in closed container and top of the percolator apparatus is closed. The
mixture is allowed to stand for 24 h. The outlet of the percolator then is opened and the liquid
contained therein were allowed to drip slowly. Later sufficient solvent is added and the mixed
liquid is clarified by filtration or by standing followed by decanting [14].
Fig 3: Stas Otto process (method-1)
Fatty component removed by
sodium carbonate, lime,
ammonia
Extract by ether, chloroform with
H2SO4
Ammonia added to the
aqueous layer
Residue extracted by
chloroform in percolator apparatus
Distilled off under vacuum
![Page 15: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/15.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 10
4.6. Stas otto process (Method-2)
The Powdered plant material was acidified with tartaric acid and then extracted with
Ethanol (90-95%). The alcohol portion was then distilled off under vacuum condition. Later
Petroleum ether was added to the aqueous residue to remove fatty components. Again the
aqueous portion was filtered and evaporated in Rotary evaporator [15]
.
Fig 4: Stas Otto process (Method-2)
Aqueous layer
Residue extracted with ethanol and Alkaloids precipitated
Acidified sample with tartaric acid
Ethanol added (90-95%)
Alcohol distilled off under vacuum Aqueous layer
Petroleum ether added (60-80°C)
Fatty component removed
If any alkaloid removed by
petroleum ether recovered by dil.
mineral acids
Filtered and evaporated to dryness in
rotary thin film evaporation
![Page 16: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/16.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 11
4.7. Kippenberger’s process
In the Kippenberger’s process, the powdered sample was digested with tannin in glycerol
at 40°C for 48hr. The residue was again heated, then cooled and filtered. To the filtrate the
petroleum ether was added to remove all the fatty components. Then the petroleum ether was
removed either on electric water bath or exposure to IR lamp. The fat free part was then acidified
and shaken well with the chloroform to remove the narcotine, codeine alkaloids. And CO2 was
passed to remove morphine, narceine. Alkali hydroxide converts into carbonate and finally
subjected to alcohol and chloroform. Third alkaloids are removed by equal volume of ether and
chloroform [15].
Fig 5: Kippenberger’s process
CO2 passed. Extracted using alcohol and chloroform
Acidified and shaken with chloroform
Powdered sample
Digest with tannin in glycerol at 40°C for 48 h
Residue heated at 50°C cooled and filtered
Filtrate shaken with petroleum ether
Fatty material removed Extract heated in water bath
(electric) or exposure to IR-lamp
Petroleum ether removed Fat free crude
Third alkaloids extracted using ether and
chloroform. And crude alkaloids precipitated.
![Page 17: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/17.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 12
4.8. Maceration
The coarsely powdered plant material was soaked in suitable solvent and kept in closed
container for a defined period with agitation to remove all soluble components dissolved in it.
After few hours, the marc was then filtered and the residues were filtered using Whatman filter
paper. On repeated maceration, the yield extracted will be very high effectively. Maceration
extraction is best method for the thermo-labile alkaloids. The maceration method is time and
solvent consuming [17]
. In recent years, there is new emergence of maceration methodology
where, unorganized drugs such as resin, gum, etc., were carried out [17]
. The different methods of
maceration were listed below.,
Powdered plant
material
Taken in a closed
container
80% Ethanol was
added
Placed for seven days
Residue pressed
Filtered using
whatman paper
Maceration process repeated
Residue filtered
Solvent distilled &
concentrated
• Useful for unorganized drugs
• Quick process and no need to press the marc
Modified maceration
• Generally not used
• Maximum efficiency
Multiple maceration
• Shorter time extraction
• Increase permiability of cell walls
Vacuum maceration
Scheme 6: Types of maceration
extraction
Scheme 7: Maceration extraction
![Page 18: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/18.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 13
4.9. Manske’s process
The samples were defatted with petroleum ether and alcohol was added to the residue. The
alcoholic extract was dissolved in water, the pH were adjusted to 2 and then filtered. To the
filtrate ether and chloroform was added, the non-alkaloidal impurities were removed and the
organic layer was evaporated and crude alkaloids were collected [18]
.
Organic layer taken and evaporated
Sample and petroleum ether
Residue and methanol
Alcoholic extract (PH to 2)
Steam distillation
Placed in refrigerator several
days (impurities removed)
Filtered and ether,
chloroform added
Non-alkaloid impurity removed
and basified, treated with
chloroform or ether
Organic layer taken and
evaporated
Crude alkaloids
Heated with paraffin
Filtered and ether, chloroform added
Non-alkaloid impurity removed and
basified then treated with chloroform
or ether
Crude alkaloids
Defatted
![Page 19: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/19.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 14
4.10. Negative pressure cavitation extraction (NPCE)
About 25 g of the powdered sample was transferred into the extraction chamber with
different concentration of extracting solvent. A vacuum pump was connected to the extraction
pot through a condenser to generate negative pressure. The pressure was monitored by adjusting
the airflow rate this was supplied from the bottom of the device. After each extraction, the
extraction solutions were combined and dried on a rotary evaporator at 45°C. The HPLC-grade
methanol was added in a constant volume to prepare the samples for RP-HPLC analysis [19]
. This
method offers many advantages namely, low cost, reduced noise, low energy, short time
extraction, high yield and Eco-friendly.
Fig 6: Manske’s process
Sample and solvent is taken in
extraction pot
Negative pressure device
connected to vacuum pump
Vacuum pump connected
with condenser
Nitrogen gas passed into
extraction pot
After extraction solvent
collected into collection pot
Residue collected from pot
Negative
pressure
cavitation
extraction
Fig 7: Negative pressure cavitation extraction
![Page 20: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/20.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 15
4.11. Microwave-assisted extraction (MWE)
The electromagnetic radiation consists of electric and magnetic field perpendicular to each
other. The microwave generated in the frequency of 300MHz to 300GHz. This microwave
extraction based on the two mechanisms namely, ionic conduction and dipole rotation. The
migration of ions under electric field is called as ionic conduction. When the solution is
resistance to the migration of ions, then friction is generated and the solution is heated.
Therefore, dipole of the molecules changes under electric field and is known as dipole rotation.
The heating occurs only at 2450 MHz. microwave interacts with polar solvent and heating occur
due to the influence of above two phenomena [20]-[22]
.
Residue transferred into 10 ml volumetric flask and chloroform
added for further analysis
Sample and solvent
Placed in microwave extraction system MES 1000
Extract allowed cooling to room temperature before
opening the vessel
Extract filtered through anhydrous sodium sulphate into 100 ml
RB flask
Solvent evaporated by rotary vacuum evaporated at 40°C
Fig 8: Microwave assisted extraction
![Page 21: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/21.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 16
4.12. Accelerated solvent extraction (ASE) or pressurized solvent extraction (PSE)
The accelerated solvent extraction method is a new technique which holds, less solvent
requirement, single stage extraction, shorter time, and the faster kinetics when compared with
other methods. The main advantage of this method is the high temperature and pressure
improves metabolite solubilization, decrease in liquid viscosity and accelerate the extraction
kinetics which leads to the high yield [23].
Sample, Sea sand (4:1) loaded
in ASE 100ml extraction vessel
Solvent filled in reservoir bottle and
Extracted (70-100°c) at 1000psi to 2000 psi
Solvent transferred into 25 ml flask
Evaporated at 50° c by rotary evaporated
Residue dissolved in 25 ml of water
Filtered through 0.22µm filter
Nitrogen
atm.,
Fig 9: Accelerated solvent extraction
![Page 22: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/22.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 17
4.13. Subcritical water extraction (SWE)
The Extraction of phytochemicals using hot water (100 to 250°C) under pressure is known as
subcritical water extraction. This method works on the bases of dielectric constant of water with
temperature. At room temperature dielectric constant of water nearly is close to 80. When water
heated up to 250°C dielectric constant becomes 27. Here Reservoir connected to pump to
introduce solvent into the system [24]
.
![Page 23: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/23.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 18
Fig 10: Subcritical water extraction
4.14. Ultrasonic assisted extraction (sonication)
The Sample was transferred into glass container in contact with 50ml of solvent and closed
with aluminum foil then placed in ultrasonic bath. The Ultrasonic extraction works from 20 kHz
which induce a mechanical stress to produce cavitation on plant material. The High yield
obtained in an increase of solubilization of metabolite by breaking of cellular. The extraction
was repeated for every 30min and extract was filtered with anhydrous sodium sulphate and the
solvent was evaporated in rotary evaporated at 45°C [25]
.
Sample loaded into cell
Cell filled with solvent at 1500 psi pressure
Initial heat up time applied
Static extraction performed with all system valves closed
Cell is rinsed with 60% volume of cell using solvent
Solvent purged from cell by Nitrogen gas
Depressurization
Solvent evaporated
Extract collected and protected from
light at 4°C
Residue
![Page 24: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/24.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 19
Fig 11: Ultrasonic assisted extraction
4.15. Pulse electric field extraction (PEFE)
The principle of Pulse Electric Field Extraction is to destroy the cell membrane structure to
increase the kinetics of the extraction. During the suspension of a living cell in an electric field,
the electric potential passes through the membrane of that cell. Based on the dipole nature of
membrane molecules, the electric potential separates the molecules according to their charge in
the cell membrane. The PEF can increase the mass transfer during the extraction by destroying
the membrane structure of plant materials for enhancing extraction and decreasing extraction
time. The dried sample was mixed with solvent and soaked for 24 h. The mixture was pumped
through the PEF system with a flow velocity of 120 L h-1. After being processed for the desired
time, the high voltage pulsing was turned off, the mixture was pre-filtered through a piece of
tulle to remove solid particulates, and then through a 0.45-µm hydrophilic polypropylene
membrane [26]
.
Sample taken in glass container
Suitable solvent added
Put the container
in ultrasonic bath frequency in 40kHz at 40°C
extraction repeated and crude alkaloids collected
![Page 25: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/25.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 20
Fig 12: Pulse electric field extraction
4.16. Merck process
In the Merck process, the sample was mixed with cold water and soaked for 24h. The liquid
obtained is concentrated and precipitated hot with sodium carbonate. Ammonia is then
evaporated on heating the mixture and it should be alkaline to phenolphthalein. After 24h the
mixture is filtered, dissolved in alcohol at 85 °C. The excess alcohol is then evaporated and
residue mixed with acetic acid for neutralization. This acetic solution is then decolorized with
charcoal, filtered and precipitated with ammonia [27]
.
Sample in cold water
Sodium carbonate added and heated, left for 24h
Precipitate is filtered and washed with cold water
Precipitate dissolved in alcohol at 85°C
Sample soaked in solvent for 24h
Mixture pumped through PEF
system at 120 Lh-1 velocities
Pulse frequency, charge voltage adjusted to the desired level and
extraction strated
Filtered through 0.45µm
hydrophilic polypropylene
![Page 26: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/26.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 21
4.17. Thiboumery and Mohr process
The sample material was treated with hot water with triplicate. The liquid is filtered off
and the residues were again treated with three times its weight of water. The solutions evaporated
to half their volume and poured into boiling milk of lime. The precipitate were filtered off and
re-treated with three parts of water to one part of opium. It is then filtered off again and the
solution was heated to boiling, and the alkaloids were precipitated by the addition of ammonium
Fig 13: Merck process
![Page 27: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/27.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 22
chloride. After cooling, it is filtered and the precipitate was washed and then purified by solution
in hydrochloric acid [28]
.
4.18. Robertson-Gregory process
The plant materials were completely treated with five to ten times of cold water
and the solution was evaporated and the process was repeated with cold water. The solution
obtained was evaporated and few crystals of calcium chloride added to the boiling solution
which is then diluted with cold water. The solution was then filtered and concentrated further.
This deposit is filtered off and the solutions were left in on-standing. After a few days, the
Fig 14: Thiboumery and Mohr process
Sample and hot water (3times than sample)
Filtered and residue retreated with hot
Evaporated and poured into boiling milk
Precipitate retreated with hot
Filtered off again, ammonium chloride
Filtered, heated to boiling and cooled. Again filtered
Precipitate washed and purified by HCl
![Page 28: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/28.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 23
filtrated material it becomes a crystalline in nature. The crystals obtained are drained and then
placed in a cloth. The solution were then decolorized each time with animal charcoal and
dissolved in water and the alkaloids are precipitated with ammonia [28]
.
`
5. Separation of different types of alkaloids [29]-[30]
Sample and cold water (5times than sample)
Solution evaporated and retreated with cold water
Calcium chloride added to the boiling solution
Diluted with cold water and concentrated
Filtered off and left for few days.
Alkaloids in acidic solution and filtered
Extract with organic solvent (methylene chloride)
Dichloromethane layer
Extract with 1-2% NaOH
Acidic layer taken and add
ammonia dichloromethane
Fig 15: Robertson-Gregory process
![Page 29: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/29.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 24
Fig 16: Separation of Alkaloids (A)
![Page 30: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/30.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 25
Fig 17: Separation of Alkaloids (B)
6. Qualitative analysis of alkaloids
The qualitative analyses of alkaloids are important study for the quantification of alkoaids
in a particular crude extract. The concentrated extract solution was used for the investigation of
alkaloids for the lack of satisfactory crystals formation. When alkaloids or alkaloids salts were
not dissolved in water, 10% HCl was used to dissolve the crystals. These solutions are taken for
the test with different reagent. The crystals formed are visible under microscopic condition. Few
drops of test solution and reagent added on glass rod and examine for microscopic condition.
Sometimes the crystals not identified quickly. Hence care should be taken and stir the
solution to form crystals. The stirring with glass rod is avoided on the account of abnormal result
of crystals. Crystal shows variation upon amount of reagent added. Normally few ml of test
solution were needed to the formation of crystals. The following table consists of some chemical
test for alkaloids. Different class of alkaloids gives different color with reagents.
Types of Separation of
Alkaloids
based on pH
based on Solubility
based on Functiona
l group
Chromato
-graphy
HPLC
![Page 31: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/31.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 26
Table 1 Chemical test for Alkaloids [31]-[36]
S.no Experiment name Chemical required Inference
1 Mayer’s Test 1.36g of Mercuric chloride, 3g of Potassium
iodide in 100 ml of Water
Cream color
Precipitate
2 Wagner’s Test 1.3g of iodine and 2g of potassium Iodide Brown or Reddish
Color precipitate
3 Kraut’s Test 8g of Bismuth Nitrate, 20ml of Nitric acid
and 27.2g of Potassium Iodide in 100ml of
Water
Brown color
precipitate
4 Marme’s Test 10g of cadmium Iodide boiled in 100ml of
Water then 20g of Potassium Iodide added
White color
precipitate
5 Scheiblers Test 20g of sodium tungstate, 70g of Disodium
phosphate in 100ml of water
White color
precipitate
6 Hager’s Test Saturated Picric acid Yellow color
precipitate
7 Sonnenschein
reagent test
1% solution of Phospho molybdic acid in
Ethanol
Orange or Red
color precipitate
8 Bertrand’s test Silicotungstic acid in distilled water
White color
precipitate
9 Reineckate salt
solution
1g of Ammonium Reineckate, 0.3g of
Hydroxyl Amine Hydrochloride and 100ml
of Ethanol
Pink color
precipitate
10 Froehde reagent
Test
5g of Molybdic acid or 5ml of Sodium
Molybdate in 5ml of concentrated Sulphuric
acid
Blue-Green color
precipitate
11 Erdmann’s reagent
Test
10drops of con. Nitric acid, 100 ml of water
is added to 20ml of con. Sulphuric acid.
Cream color
precipitate
12 Marquis reagent test 3drops of Formaldehyde (40%) and 3ml of
Sulphuric acid.
Orange-Green
![Page 32: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/32.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 27
13 Mandelin’s Reagent
Test
1g of Ammonium vanadate, 100ml of con.
Sulphuric acid.
Olive black
14 Mecke’s reagent
Test
1g of Selenious acid mixed with 100ml of
con. Sulphuric acid
Red color
precipitate
15 Rosenthaler Reagent
Test
1g of Potassium Arsenates in 100ml of con.
Sulphuric acid
Red color
precipitate
16 Folin-ciocalteu’s
phenol Test
Folin reagent and equal volume of distilled
water
White color
precipitate
17 Van Urk reagent
Test
Para Dimethyl Aminobenzaldehyde in 65%
of Sulphuric acid or HCl
Reddish brown
color precipitate
18 Dragendorff’s
reagent test
Potassium Bismuth Iodide Reddish Brown
color precipitate
Test for Aconitine [37]
1. Amorphous precipitate from in the solution of sodium carbonate
2. Slowly crystals form with potassium permanganate.
3. Slowly crystals form with potassium hydroxide
Test for Anhalonine [37]
1. Precipitate formed with Kraut’s reagent(1:200)
2. Stirring with 1:50 solution of Gold chloride a large rods are formed
3. Amorphous precipitate formed with picric acid. With more dilute solution we get small
solids.
4. Alkaloids with Wagner’s reagent form a heavy amorphous precipitate.
Test for Brucine
1. With concentrated solution of potassium chromate rosettes are formed.
2. Small crystals are formed with Platinum chloride.
3. Many of the needles formed with Potassium ferricyanide.
4. Million’s reagent gives a rose-red color with alkaloids.
![Page 33: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/33.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 28
Test for Caffeine
1. Needle shaped cyrstals formed with Mercuric chloride.
2. Very small and amorphous precipitates formed with Silico-tungstic acid.
Test for Coumarins [38]
1. To the alkaloid 1N sodium hydroxide added, test tube covered with filter paper and heated
in water bath and examined for UV light for yellow fluorescene.
2. Add few ml of chloroform and 10% sodium hydroxide yellow color formed.
Test for Choline
1. Heavy Amorphous precipitate formed with phosphortungstic acid.
2. A large number of small crystals formed with Barium nitrate.
Test for Cocaine Hydrochloride
1. Purple colored plates are formed with Potassium Permanganate.
2. Yellowish precipitate formed with Chromic acid and a small amount of concentrated
Hydrochloric acid.
3. Ferric chloride added to the alkaloids and stirred well. A blade like crystals formed with
chisel shaped ends.
Test for Codeine
1. Amorphous precipitate formed with Marme’s reagent.
2. Yellow color precipitate obtained by adding Million’s reagent
Test for Heroine (Diacetyl Morphine)
1. Rosettes of rods obtained with sodium phosphate.
2. Rosettes are formed with 1:200 solution of potassium cyanide.
Test for Hyoscyamine
1. Vitali-morin’s test: Alkaloid treated with fuming Nitric acid and evaporated to dryness.
To the residue add acetone
![Page 34: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/34.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 29
Test for Morphine
1. Marme’s reagent: this is the best test for morphine. Gelatinous precipitate formed with
1:1000 of reagent.
2. Wagner’s reagent: a reddish brown precipitate formed before crystallization takes place.
3. yellow brown shiny crystals are formed with zinc chlor-iodide
4. Alkaloids form a needle in 1:50 solution of potassium iodide.
Test for Nicotine
1. It forms an amorphous precipitate with Kraut’s, Wagner’s Mayer’s reagent.
2. Small crystals are obtained with Barium chloride.
3. With more concentrated solution of Mercuric chloride forms an Amorphous precipitate.
Test for Quinoline
1. An Amorphous precipitate formed with Wagner’s, kraut’s and Mayer’s reagent.
2. Light-pink color formed when 1:50 solution of Million’s reagent added.
Acknowledgement
The authors wish to acknowledge DST-SERB (DST-FTYS-SB/FT/CS-113/2013) for providing
funding to carry out our research. Also, we wish to thank VIT University for providing
infrastructure.
![Page 35: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/35.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 30
Fig 18: Soxhlet extraction Fig 19: Microwave assisted extraction.
Fig 20: Ultrasonic assisted extraction Fig 21: pressurized solvent extraction
x
sealed glandto part of infused
valve closed
vaccum regulator
vaccum pump
degassed resin
![Page 36: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/36.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 31
Fig 22: Percolation method Figure 23: Heat reflux
Fig 24: Rotatory evaporator
![Page 37: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/37.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 32
References
[1]. Doughari JH.2012, Phytochemicals: Extraction Methods, Basic Structures and Mode of
Action as Potential Chemotherapeutic Agents. Department of Microbiology, School of Pure and
Applied Sciences, Federal University of Technology, Yola, Nigeria, 1-32.
[2]. Khan AM, Quresh RA, Ullah F, Gilani SA, Nosheen A, Sahreen S, Laghari MK, Laghari
MY, Shafiq-Ur R, Hussain I, Murad W.2011. Phytochemical Analysis of Selected Medicinal
Plants of Margalla Hills and Surroundings. J. Med. Plants Res. 5(25): 6017-6023.
[3]. Cordell G.A. 1983, Introduction to Alkaloids, A Biogenic approach,wiley, new york
[4]. Robert M.F and Wink M. 1998. Alkaloids: Biochemistry, Ecology and medicinal
application. Plenum press, New York.
[5]. Shankul kumar. 2010. Opium alkaloids
[6]. Hesse, Manfred (2002). Alkaloids: Nature's Curse or Blessing. Wiley-VCH. ISBN 978-3-
906390-24-6.
[7]. Handa SS, Khanuja SPS, Longo G, Rakesh DD.2008. Extraction Technologies for Medicinal
and Aromatic Plants. International centre for science and high technology, Trieste,
21-25.
[8]. B. Benthin, H. Danz and M. Hamburger, 1999. J. Chromatogr., A, 837, 211–219.
[9]. Luque de Castro M.D, Priego-Capote F. Soxhlet extraction: Past and present panacea.
Journal of Chromatography A, 1217 (2010) 2383–2389
[10]. Behr labor the extraction procedure www.behr-labor.com
[11]. Nuat duoc lieue. 2012. General methods of extraction and isolation of alkaloids. July 7.
![Page 38: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/38.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 33
[12]. Sunil kumar. 2014. Alkaloidal drugs-A review. Asian Journal of pharmaceutical science
and technology
[13]. Ncube NS, Afolayan AJ, Okoh AI. 2008. Assessment techniques of antimicrobial
properties of natural compounds of plant origin: current methods and future trends. African
Journal of Biotechnology, 7 (12): 1797-1806.
[14]. Jain N.K and Sharma. Text book of professional pharmacy, 115.
[15]. Shankul Kumar, 2011. Extraction of phytoconstituent.
[16]. Li-li dong, Yujie Fu, yuan-gangzu. August 2011. Negative pressure cavitation accelerated
processing for extraction of main bioactive flavonoids from Radix Scutellariae, chemical
engineering and processing: process intensification, 50(8), 780-789
[17]. Bethe H.A. 1947. The electromagnetic shift of energy levels. Phy.rev.72. 339-341.
[18]. Anastassiades J.D, Kaminsky E.H, Perea and Poezevara A. 1992. Solid-state microwave
generation, microwave technology series. Vol.4. Chapman and Hall London.
[19]. Carroll J.E. 1970. Hot electron microwave generators. Edward Arnold, London.
[20]. Haiyan wang, yuchaolu, jie chen, junchao, li jhuhui liu. 2012. Subcritical water extraction
of alkaloids in Sophora Flavescens Ait and determination by capillary electrophoresis with field
amplified sample stacking, journal of pharmaceutical and biomedical analysis, 58, 146-151.
[21]. Miguel Herrero, Alejandro Cifuentes, Elena Iban ez. 2006. Food chemistry: Sub- and
supercritical fluid extraction of functional ingredientsfrom different natural sources: Plants, food-
by-products, algae and microalgae. Vol.98. 136-148.
![Page 39: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/39.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 34
[22]. Lan-Sook Lee, Namhyouck Lee, Young Ho Kim, Chang-Ho Lee, Sang Pil Hong, Yeo-Won
Jeon and Young-Eon Kim. 2013. Molecules: Optimization of Ultrasonic Extraction of Phenolic
Antioxidants from Green Tea Using Response Surface Methodology, 18, 13530-13545.
[23]. Azmir J, Zaidul I.S.M, Rahman M.M, Sharif K.M, Mohamed A, Sahena F, Jahurul
M.H.A, Ghafoor K, Norulaini N.A.N, Omar A.K.M. 2013. Techniques for extraction of
bioactive compounds from plant materials: A review. Journal of Food Engineering. 117,
426–436.
[24]. Andre barbier, the opium alkaloids extraction.
[25]. Robert J Bryant. 1987. The manufacture of medicinal alkaloids from the opium poppy.4-6.
[26]. Soni B, Visavadiya NP, Madamwar D. 2008. Toxicology, 248(1), 59-65.
[27]. Yubin JI, Yu Miao, Wang Bing and Zhang Yao. 2014. The extraction, separation and
purification of alkaloids in the natural medicine. Journal of Chemical and Pharmaceutical
Research, 6(1), 338-345.
[28]. James Hamuel Doughari, phytochemical: Extraction methods, Basic Structure and mode of
action as potential chemotherapeutic Agents.
[29]. Balakrishnan Arunachalam, Kokilavani Chinnaraju. 2012. preliminary phytochemical
analysis and cytotoxic potential of Cucumins Trigonus Roxb, journal of intercult
Ethnopharmacol, 1(1), 13-18.
[30]. Sunil kumar. 2014. alkaloidal drugs - a review, Asian journal of pharmaceutical science and
technology,4(3), 107-119.
[31].Arti Dixit, Ankur Rohilla, Jyoti Dixit and Vijender Singh. 2014. qualitative analysis of
various plant extracts of Alpinia Officinarum, international journal of pharmaceutical, chemical
and biological sciences, 4(3), 505-508.
![Page 40: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/40.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 35
[32]. Ashutosh kar. 2003. pharmacognosy and pharmacobiotechnology, 448-451.
[33]. S.B.Gokhale, C.K.Kokate, A.P.Purohit. A text book of pharmacognosy, 8.2-8.4
[34]. Nuat Duoc lieu. 2012. Ergot alkaloids,pharmacognosy.
[35]. Asia Nosheen.2011. Phytochemical analysis of selected medicinal plants of Margalla Hills
and surroundings, journal of medicinal plants research, 5(25),6017-6023.
[36]. K.N.Shailesh Kumar and S.Hemalatha. 2013. Phytochemical evaluation of leaf extracts of
Aegle marmelos, International Journal of Development Research, 3(7),29-33.
[37]. Charles H. Stephenson and C.E.Parker, some micro chemical tests for alkaloids.
![Page 41: Corrected hand book - ISCA 978-93-84648-67-1.pdf · A HAND BOOK ON: SEMI MICRO ... This handbook provides an easy and brief ... long time extraction, solvent consumption, less efficiency](https://reader031.vdocuments.mx/reader031/viewer/2022021823/5b3621097f8b9aad388cb4d6/html5/thumbnails/41.jpg)
International Science Congress Association
www.isca.in , www.isca.co.in , www.isca.net.co , www.isca.net.in
A HAND BOOK ON: SEMI MICRO TECHNIQUE FOR EXTRACTION OF ALKALOIDS 36
ABOUT THE AUTHORS
Dr G. Madhumitha is presently working as Assistant Professor,
Organic Chemistry Division, School of Advanced Sciences in VIT
University, Vellore. She is teaching Unit Process in Organic
Synthesis, Environmental Studies and Engineering Chemistry to UG
students since last two years. She did her B.Sc. degree in Applied
Chemistry from CIT University, Coimbatore; M.Sc., M.Phil., Ph.D.
from VIT University, Vellore and M.BA. From Bharathiyar
University, Coimbatore. She has published more than 15 research papers in international
journals of repute. She has received Young Scientist Award (FTYS-Scheme) from Department of
Science and Technology, New Delhi. Past 2 years she has received Research Award from VIT
University. Currently six research scholars are doing their Ph.D. work under her. Also she is
guiding B.Tech. and M.Sc. students.
J. Fowsiya has completed her Under Graduate degree (2008-11) in
chemistry in Muthurangam arts college, Vellore and master degree in
chemistry (2011-13) in Guru Nanak College, Velacheri, Chennai. She had
one year of teaching experience in S.S.S. arts and Science College, Arcot,
Vellore. In the duration of teaching days She handled physical chemistry,
general chemistry and spectroscopy and recorded a good teaching and result
in S.S.S. college. Now she is working as research assistant from Sep 2013 in VIT University
under the guidance of Dr. G. Madhumitha, Asst. Prof, Organic chemistry division, VIT
University, Vellore-14. She thank to almighty Allah for given strength to my entire life and to her
guide Dr. G. Madhumitha, Asst. Prof for giving a good opportunity to make her dreams becomes
true and also thankful to all her School, UG, PG and her parents and friends.