Definition, Classification and Properties of Alkaloids:

Definition

Any of a class of naturally occurring organic nitrogen-containing bases. Alkaloids

have diverse and important physiological effects on humans and other animals. Well-known
alkaloids include morphine, strychnine, quinine, ephedrine, and nicotine. Alkaloids are found
primarily in plants and are especially common in certain families of flowering plants. In fact,
as many as one-quarter of higher plants are estimated to contain alkaloids, of which several
thousand different types have been identified. In general, a given species contains only a few
kinds of alkaloids, though both the opium poppy (Papaver somniferum) and the ergot
fungus (Claviceps) each contain about 30 different types. Certain plant families are
particularly rich in alkaloids; all plants of the poppy family (Papaveraceae) are thought to
contain them, for example. The Ranunculaceae (buttercups), Solanaceae (nightshades), and
Amaryllidaceae (amaryllis) are other prominent alkaloid-containing families. A few alkaloids
have been found in animal species, such as the New World beaver (Castor canadensis)
and poison-dart frogs (Phyllobates).[1]

Key characteristics of alkaloids include:

1. Nitrogen Content: Alkaloids contain nitrogen atoms, often in a cyclic structure. The
nitrogen is typically in a basic or alkaline form.
2. Natural Occurrence: Alkaloids are found in nature, primarily in plants. However, some
are produced by fungi and a few by animals.
3. Biological Activity: Many alkaloids have notable physiological effects on living
organisms. Some alkaloids exhibit pharmacological properties and can be used for
medicinal purposes.
4. Diverse Structures: Alkaloids have diverse chemical structures, and they can be
classified into different groups based on their chemical characteristics.
5. Roles in Plants: In plants, alkaloids often serve as secondary metabolites. They may act
as defense mechanisms against herbivores or play a role in attracting pollinators.
6. Pharmacological Significance: Some alkaloids have been utilized in traditional medicine,
and many continue to be important in the development of pharmaceutical drugs.
Examples include morphine, quinine, and caffeine.
Different types of alkaloids:

Table: Types of alkaloids [2]

Physical Properties:
The physical properties of alkaloids can vary widely due to the diverse structures within this
group of compounds. However, some general characteristics and physical properties can be
identified:

 State: Alkaloids can exist in various physical states, including solids, liquids, and, in
some cases, gases.Many alkaloids are crystalline solids but few are amorphous solids
(Example: Emetine). Solid nature due to the presence of O-atom in the ring structure.
 Solubility: Alkaloids are insoluble in water but soluble in most organic solvents.
Caffeine, cocaine, codeine, nicotine are slightly soluble in water, Morphine, yohimbine
is very slightly water-soluble.
 Optical Activity: Alkaloids are optically active. Most of the alkaloids are levo-rotatory
but few are dextro-rotatory (e.g. Coniine) and few are even optically inactive, viz.
papaverine.
 Taste: Alkaloids are bitter and have pronounced physiological activity.
 Molecular weight: They have a molecular weight of between 100-900.
 Volatility: They are sometimes non-volatile liquids (Examples: Hyoscine, Pilocarpine),
sometimes volatile liquids (Examples: Nicotine, Coniine).
 Color: They are generally colorless (Few are colored such as Colchicine and Berberine
are yellow, Betanidine is orange, Salt of Sanguinarine is cupper red).
 Melting Point: The melting points of alkaloids can vary considerably. Different alkaloids
have different temperature ranges at which they change from a solid to a liquid state.

Chemical Properties:
The chemical properties of alkaloids are diverse, reflecting the wide range of structures within
this group of naturally occurring compounds. Here are some general chemical properties
associated with alkaloids:

 Basic Nature: Alkaloids are characterized by their basic nature due to the presence of
nitrogen atoms, often in a cyclic structure. The basicity is a result of lone pair electrons
on the nitrogen atoms.
 Amphoteric Behavior: Alkaloids can behave as both acids and bases, depending on
the reaction conditions. In acidic environments, they can act as bases by accepting
protons, while in basic environments, they may act as weak acids by donating protons.
 Reaction with Acids: Alkaloids react with acids to form salts. This is a common reaction
that can be utilized in the isolation and purification of alkaloids.
 Isomerism: Alkaloids may exhibit various types of isomerism, including structural
isomerism and stereoisomerism. The presence of chiral centers in many alkaloids
contributes to their stereoisomeric forms.
  Oxidation and Reduction: Some alkaloids can undergo oxidation or reduction
reactions. These reactions can lead to the formation of new compounds with different
chemical properties.

 Complex Ring Systems: Alkaloids often contain complex ring structures, which
contribute to their diverse chemical behaviors. The presence of multiple functional
groups within these ring systems can result in a variety of reactions.

 Hydrolysis: Some alkaloids are susceptible to hydrolysis, particularly under acidic or

basic conditions. This can lead to the cleavage of bonds within the alkaloid molecule.

 Reaction with Precipitating Reagents: Alkaloids can often be identified or separated

based on their reactions with specific reagents. For example, alkaloids containing the
nitrogen atom in a heterocyclic ring may form precipitates with certain reagents.

 Synthesis: While alkaloids are primarily derived from natural sources, they can also be
synthesized in the laboratory. The synthesis of alkaloids involves complex organic
chemistry reactions.
 Biological Activity: Many alkaloids exhibit pharmacological properties due to their
interactions with biological systems. Their chemical structures often contribute to
specific biological activities, making them important in the development of drugs.

Importance in Pharmacology and Medicine

Alkaloids play a pivotal role in pharmacology and medicine due to their diverse
pharmacological activities and potential therapeutic applications. The significance of alkaloids
in these fields is underscored by their historical use in traditional medicine and their continued
contribution to modern drug discovery. The importance can be elaborated through several
key points:
 Biological Activity and Therapeutic Potential: Alkaloids often exhibit potent biological
activities that can be harnessed for therapeutic purposes. Their structural diversity
allows them to interact with various biological targets, making them valuable
candidates for drug development.
 Analgesic and Anesthetic Properties: Certain alkaloids, such as morphine derived
from the opium poppy, have strong analgesic properties. Morphine and its derivatives
are essential components in pain management, providing relief in various medical
contexts.
 Antimicrobial Effects: Many alkaloids possess antimicrobial properties, making them
valuable in the treatment of infectious diseases. Examples include quinine (used
against malaria) and berberine (with antibacterial and antifungal properties).
 Anticancer Agents: Several alkaloids have demonstrated anticancer activity.
Vinblastine and vincristine, derived from the Madagascar periwinkle, are used in the
treatment of certain cancers due to their ability to inhibit cell division.
  Cardiovascular Effects: Alkaloids like reserpine have been used in the treatment of
hypertension. These compounds can impact cardiovascular function, making them
important in managing cardiovascular disorders.
 Central Nervous System Effects: Alkaloids influence the central nervous system,
leading to their use in treating neurological disorders. Examples include atropine and
scopolamine, which affect cholinergic receptors and find application in various medical
scenarios.
 Traditional Medicine and Ethnopharmacology: Alkaloids have been integral to
traditional medicine practices in various cultures. The knowledge of alkaloid-
containing plants has been passed down through generations, contributing to the
development of ethnomedicine and providing a basis for drug discovery.
 Drug Discovery and Development: Alkaloids continue to be a valuable source of lead
compounds in drug discovery. The unique structures and diverse bioactivities of
alkaloids make them attractive starting points for developing new pharmaceuticals.
 Toxicology and Poisoning: While alkaloids can have therapeutic benefits, some can be
toxic at higher doses. Understanding the toxicological aspects of alkaloids is crucial for
avoiding adverse effects and managing cases of poisoning.[3]

Identification of Alkaloids
(a) Precipitation reactions:

 Alkaloid sample reacts with Dragendorff’s reagent (Potassium-bismuth-iodide

solution) which gives reddish-brown precipitate. Reagent Composition: Bismuth
nitrate-8.00 g, Nitric acid- 21.5 g, Potassium iodide-27.2 g, and water to make 100 ml.
 Alkaloid sample reacts with Mayer reagent (Potassium-mercuric-iodide solution)
which gives cream color precipitate. Reagent Composition: Mercuric chloride- 1.3 g,
Potassium iodide-5.00 g, and water to make 100 ml.
 Alkaloid sample reacts with Wagner reagent (Iodine-potassium-iodide solution) which
gives Brown color precipitate. Reagent Composition-Iodine: 1.3 g, Potassium iodide-
2.0 g, and water to make 100 ml.
 Alkaloid sample reacts with Hager reagent (Saturated solution of picric acid) which
gives yellow color precipitate. Reagent Composition- Picric acid- 1.0 g, water to make
100 ml.
 Alkaloids react with Valser’s test (Mercuric iodide) which gives a white color
precipitate.
 Alkaloid sample reacts with acids and gives buff color precipitate.
 Alkaloids react with picolinic acid and give a yellow color precipitate.
(b) Important Color reactions:
 Vitali Morin Test: This test is positive for Solanaceae family drugs such as Belladonna,
Datura, Henbane, Mandrake, Tobacco, etc. Test: Sample alkaloid mixed with fuming
nitric acid, evaporated to dryness. Dissolved residue in acetone and added a
methanolic solution of KOH. It produced a violet color. It indicates the presence of the
tropane group.
 Van Urk’s Test: This test is positive for Ergot alkaloid. Test: Sample reacts with para-
dimethyl amino benzaldehyde and dilutes sulphuric acid and gives blue color. It
indicates the presence of the indole group especially the clavinet group of alkaloids.
 Friend’s Test: This test is positive for opioid alkaloids. Sample reacts with Froehd’s
reagent (sodium molybdate in concentrated sulphuric acid) and forms brownish-black
color due to the presence of the Isoquinoline group.
 Thalloquin Test: This test is positive for Quinine alkaloid. Cinchona powder reacts with
bromine water in presence of strong ammonia and gives an emerald green color due
to the presence of the quinoline group.
 Rosequin Test: This test is also known as the Erythroquinine test. This test is positive
for Quinine.Test: Sample solution added with dilute acetic acid and a few drops of
bromine water. Then added a drop of a solution of potassium ferrocyanide and added
a drop of strong ammonia solution. The solution turns to red color. In this solution, a
few ml of chloroform is added then the chloroform layer became red.
 Murexide Test: This test is positive for purine derivative alkaloids, mainly Caffeine. Test:
Caffeine sample is taken in China dish and added potassium chlorate and dilute HCl.
Evaporated to dryness. It produces a red color. Further, exposing the China dish to
 strong ammonia vapor. The red color is converted into purple or violet color. When
caffeine reacts with HCl it forms tetramethylalloxanthine which further in presence of
ammonia forms ammonium salt of tetramethylpurpuric acid (Murexide).[4]

Extraction of Alkaloids
 Stas-Otto method:

The method is based on the distribution of alkaloidal bases between acid or aqueous
solution and immiscible organic solvent. Powdered drug is mixed with water and alkali to
make a paste. Alkali is added because it is mixed with acids, tannins and other phenolic
substances to make it free of alkaloids. The thereafter mixed drug is extracted with
chloroform or other organic solvents using the Soxhlet apparatus. This step is carried out
because freed alkaloids are dissolved together with other substances soluble in the
solvent. Then further re-extract the chloroform layer with dilute sulphuric acid in the
separating funnel and the acid layer is collected. This is because alkaloids are converted
into alkaloidal sulphate which is soluble in water. Then in the acid layer water is added so
that soluble constituents are passes into the aqueous solution. Then the aqueous layer is
mixed with ammonia to make the media alkaline. Then evaporate to dryness and the
precipitate is collected and dried. This step is carried out because ammonia converts
alkaloidal sulphate to ammonium sulphate which is soluble in water. The free alkaloids
which are insoluble in water, are precipitated.
 Plant sample extracted with water or alcohol. The aqueous layer is further extracted
with an organic solvent like chloroform to remove pigments or other impurities which
are soluble in that solvent. Then shake with acid to form a salt of alkaloid in the
aqueous layer. The aqueous layer is then treated with ammonia to get freed alkaloid
which is separated with organic solvents. After evaporation to dryness residue of
alkaloidal extract forms [5].
Purification and Analysis of Alkaloids
Purification:

This technique capitalizes on the varying solubilities of alkaloids in organic solvents versus
water. Following extraction, the solvent is evaporated, yielding a crude extract that can
undergo further purification steps. Steam distillation is particularly useful for volatile alkaloids
with lower boiling points. This method utilizes steam to carry volatile compounds from the
plant material, and the resulting steam-alkaloid mixture is condensed and collected.
Supercritical fluid extraction, utilizing supercritical carbon dioxide,Solvent extraction is a
fundamental method employed for the isolation of alkaloids from plant material provides a
selective, efficient, and environmentally friendly alternative for obtaining alkaloids, especially
those that are thermally labile.

Column chromatography, a widely adopted technique, facilitates the purification of alkaloids

based on their differential affinities for a stationary phase. The mixture is loaded onto a
column, and by selecting appropriate solvents and adjusting conditions, alkaloids are Liquid
Chromatography (HPLC) is an advanced form of chromatography suitable for isolating larger
quantities of alkaloids. Flash chromatography, relying on positive pressure to force the solvent
through the column, is a rapid method for efficient large-scale purification.[

Analysis:
In the realm of quantitative analysis, chromatography-based quantification is frequently
employed. High-performance liquid chromatography (HPLC) and gas chromatography (GC)
offer precise measurements by comparing peak areas or retention times with calibration
curves generated using standard alkaloid solutions. Mass spectrometry-based quantification,
coupled with chromatography, enhances accuracy and sensitivity in detecting alkaloids.
Qualitative analysis involves confirmatory tests to identify alkaloids preliminarily. Reagents like
Dragendorff's or Mayer's reagent induce specific reactions with alkaloids, resulting in
characteristic precipitates or color changes. Chiral analysis becomes imperative for
determining the stereochemistry of alkaloids. Techniques such as chiral chromatography or
chiral capillary electrophoresis aid in the separation and analysis of enantiomers, contributing
to a comprehensive understanding of alkaloid structures. These purification and analysis
methods collectively form a robust approach, allowing researchers to obtain highly purified
alkaloids for in-depth study and to quantify their presence in diverse samples. Adjustments to
these techniques are made based on the specific alkaloid under investigation and the nature
of the sample being analyzed.[6]
References
1 Yubin, J. I., Miao, Y., Bing, W., & Yao, Z. (2014). The extraction, separation and
purification of alkaloids in the natural medicine. Journal of Chemical and
Pharmaceutical Research, 6(1), 338-345.
2 Leitão, G. G., Leal, C. M., Mendonça, S. C., & Pereda-Miranda, R. (2021).
Purification of alkaloids by countercurrent chromatography. Revista Brasileira de
Farmacognosia, 1-23.
3 Dey, P., Kundu, A., Kumar, A., Gupta, M., Lee, B. M., Bhakta, T., ... & Kim, H. S. (2020).
Analysis of alkaloids (indole alkaloids, isoquinoline alkaloids, tropane alkaloids).
In Recent advances in natural products analysis (pp. 505-567). Elsevier.
4 Pelletier, S. W. (Ed.). (1983). Alkaloids: chemical and biological perspectives (Vol. 1).
New York: Wiley.
5 Verpoorte, R., & Schripsema, J. (1994). Isolation, Identification, and Structure
Elucidation of Alkaloids A General Overview. Alkaloids, 1-24.
6 Fattorusso, E., & Taglialatela-Scafati, O. (Eds.). (2007). Modern alkaloids: structure,
isolation, synthesis, and biology. John Wiley & Sons.