Assignment of CHEM 546

(Natural Product
Chemistry Techniques) on the topic of ‘’ Methods for Extraction of
Alkaloids ‘’
Submitted To: Dr. Dildar Ahmed Alavi

Submitted By: Madiha Muhammad Yousaf

Class: M. Phil Chemistry ( Semester I )

Roll No: 253080133

Date: 27-10-2023

Alkaloids
Alkaloids are a group of naturally occurring, organic compounds that contain nitrogen atoms,
typically in a cyclic ring structure. They are known for their diverse pharmacological effects and
are primarily found in plants, although they can also be found in animals and fungi. These
compounds often have basic, or alkaline, properties.

EXTRACTION OF ALKALOIDS

Extraction of Alkaloids involves the following steps:

Preparation of Plant Material:

 Choose the plant source that contains the desired alkaloids. Using the correct part of the
plant is essential, as alkaloid content can vary between roots, leaves, stems, and seeds.
 Choose a plant extract that shows a positive test for the presence of alkaloids. This can be
determined through specific alkaloid detection tests, such as Dragendorff's test or Mayer's
test.
 Remove any dirt, dust, or foreign material from the collected plant material by gently
washing it with clean water.
 After washing, allow the plant material to air-dry or use a drying apparatus to remove
excess moisture. Proper drying helps prevent the growth of molds and fungi and ensures
that the material is in a suitable condition for grinding.
 Once the plant material is dry, grind it into a fine powder. Grinding increases the
material's surface area, improving the extraction process's efficiency. The fineness of the
powder can impact the extraction yield.
 Store the powdered plant material in a cool, dry, and dark place. Make sure it is stored in
airtight containers to prevent moisture absorption and contamination. Proper storage
helps maintain the stability of the plant material.

Removal of Plant Oils:

  Use a suitable solvent that is selective for dissolving plant oils while sparing the alkaloids
or other compounds of interest. Common solvents for this purpose include hexane,
petroleum ether, or diethyl ether.
 Add the solvent to the crushed or ground plant material. The solvent should be chosen
based on its compatibility with the plant material and the specific plant oils you want to
remove.
 Agitate the mixture to ensure thorough contact between the solvent and plant material.
This can be done by stirring, shaking, or using mechanical agitation equipment.
 The solvent is then separated from the mixture using techniques such as filtration or
centrifugation.
 The residue obtained after solvent removal contains alkaloids, salts, glycosides, and other
polar compounds

Treatment with Bases

 The residue containing free alkaloids, or their salts, or glycosides are treated with bases
like Ca(OH)2, NH4OH, or Na2CO3 to liberate the free alkaloids.
 The free alkaloids may now be partitioned between water and an organic solvent like
benzene.

Extraction with Organic Solvent:

 Organic solvents such as methanol (CH3OH), ethanol (C2H5OH), or chloroform

(CHCl3) are commonly used for this purpose.
 The extraction process involves combining the prepared plant material with the solvent,
followed by agitation to facilitate the dissolution of alkaloids into the organic phase.
 After extraction, the solid plant material is separated from the solvent mixture through
filtration, leaving behind an organic solution containing the alkaloids.
 The next step involves concentrating this solution by evaporating the solvent, typically
using a rotary evaporator.

Extraction of alkaloids with acid

  The organic phase obtained from the previous step, which contains the alkaloids
dissolved in the organic solvent, is subjected to acid extraction.
 An acidic aqueous solution is prepared, utilizing weak acids like hydrochloric acid or
acetic acid. This acidified solution is combined with the prepared plant material, and the
alkaloids within the plant material are dissolved into the solution. The acidic environment
facilitates the conversion of alkaloids into their salt forms, which are more water-soluble.
 After extraction, the solid plant material is separated from the aqueous solution by
filtration, leaving an alkaloid-rich solution. To recover the alkaloids in their pure form,
the pH of the solution is adjusted to promote their conversion back into their free-base
form. Subsequently, the pH-adjusted solution is subjected to organic solvent extraction,
with the alkaloids transferring from the aqueous phase to the organic phase. The two
phases are then separated using a separatory funnel, and the organic phase, now
containing the alkaloids, is concentrated by evaporating the solvent.

Extraction with Aqueous Acids:

 Extraction of alkaloids with aqueous acids is a method commonly used to separate

alkaloids from plant material. It begins by immersing the prepared plant material in an
acidic aqueous solution, often containing weak acids like hydrochloric or acetic acid.
 The acid dissolves the alkaloids, converting them from their less soluble salt forms into
more soluble free-base forms. After thorough agitation and soaking, the alkaloid-rich
aqueous solution is separated from the solid plant material.
 The pH is then adjusted to convert the free-base alkaloids back into their salt forms, and
the alkaloids are further extracted into an organic solvent, which is immiscible with
water. The organic phase, now containing the alkaloids, is separated, and the solvent is
evaporated, leaving a concentrated alkaloid extract.

How to Obtain Individual Alkaloids?

To obtain individual alkaloids from a mixture, various chromatographic and chemical techniques
are commonly used. Here are the key methods:
 Column Chromatography: Column chromatography is often the initial step, where the
crude alkaloid mixture is loaded onto a column filled with a stationary phase. As the
mixture is eluted through the column, different alkaloids are separated based on their
chemical properties and interactions with the stationary phase. Fractions are collected,
and the target alkaloid can be isolated.
 Thin-Layer Chromatography (TLC): Thin-layer chromatography is a quick and simple
technique used for initial separation and identification. A small amount of the mixture is
spotted on a thin layer of adsorbent, and as a solvent flows up the plate, compounds are
separated based on their affinity for the adsorbent. Specific spots can be scraped off for
further isolation.
 High-Performance Liquid Chromatography (HPLC): HPLC is a high-resolution
technique used for precise separation and purification of individual alkaloids. It involves
a high-pressure liquid mobile phase passing through a column with a stationary phase,
allowing for fine separation and isolation of target compounds.
 Preparative Chromatography: Preparative chromatography involves using larger
columns and greater sample loads to isolate larger quantities of a specific alkaloid. This is
often used after initial chromatographic separation.
 Recrystallization: After isolating a compound, recrystallization can be used to further
purify it. The compound is dissolved in a solvent and then allowed to slowly crystallize.
Impurities are left in the solution.
 Spectroscopy: Analytical techniques like mass spectrometry, nuclear magnetic
resonance (NMR) spectroscopy, and infrared (IR) spectroscopy can be used for structural
identification and purity assessment of the isolated alkaloids.