Alkaloids

Definition: Definition: the term alkaloid (alkali-like) is (alkalicommonly used to designate basic heterocyclic nitrogenous compounds of plant origin that are physiologically active. active.

Deviation from Definition:

Basicity: Some alkaloids are not basic e.g. Colchicine, Piperine, Quaternary alkaloids.  Nitrogen: The nitrogen in some alkaloids is not in a heterocyclic ring e.g. Ephedrine, Colchicine, Mescaline.  Plant Origine: Some alkaloids are derived from Bacteria, Fungi, Insects, Frogs, Animals.


Classification:
True (Typical) alkaloids that are derived from amino acids and have nitrogen in a heterocyclic ring. e.g Atropine  Protoalkaloids that are derived from amino acids and do not have nitrogen in a heterocyclic ring. e.g Ephedrine  Pseudo alkaloids that are not derived from amino acids but have nitrogen in a heterocyclic ring. e.g Caffeine  False alkaloids are non alkaloids give false positive reaction with alkaloidal reagents.


New Definition: Alkaloids are cyclic Definition:

organic compounds containing nitrogen in a negative state of oxidation with limited distribution among living organisms. organisms.

Distribution and occurrence:

Rare in lower plants.  Dicots are more rich in alkaloids than Monocots.  Families rich in Alkaloids: Apocynaceae, Rubiaceae, Solanaceae and Papaveracea.  Families free from Alkaloids: Rosaceae, Labiatae


Distribution in Plant:
 All

Parts e.g. Datura.  Barks e.g. Cinchona  Seeds e.g. Nux vomica  Roots e.g. Aconite  Fruits e.g. Black pepper  Leaves e.g. Tobacco  Latex e.g. Opium

Forms of Alkaloids:
 Free

bases  Salts with Organic acids e.g. Oxalic, acetic acids  Salts with inorganic acids e.g. HCl, H2SO4.  Salts with special acids: e.g. Meconic acid in Opium Quinic acid in Cinchona  Glycosidal form e.g. Solanine in Solanum. Solanum.

Function in Plants


   

They may act as protective against insects and herbivores due to their bitterness and toxicity. toxicity. They are, in certain cases, the final products of detoxification (waste products). products). Source of nitrogen in case of nitrogen deficiency. deficiency. They, sometimes, act as growth regulators in certain metabolic systems. systems. They may be utilized as a source of energy in case of deficiency in carbon dioxide assimilation. assimilation.

Nomenclature:
Trivial names should end by "ine". These names "ine". may refer to:  The genus of the plant, such as Atropine from Atropa belladona. belladona.  The plant species, such as Cocaine from species, Erythroxylon coca. coca.  The common name of the drug, such as Ergotamine from ergot.  The name of the discoverer, such as Pelletierine discoverer, that was discovered by Pelletier.  The physiological action, such as Emetine that action, acts as emetic, Morphine acts as narcotic.  A prominent physical character, such as Hygrine character, that is hygroscopic.

Prefixes and suffixes:

Prefixes:  "Nor"Nor-" designates N-demethylation or N-demethoxylation, e.g. norpseudoephedrine and nornicotine. nornicotine.  "Apo"Apo-" designates dehydration e.g. apomorphine. apomorphine.  "Iso- pseudo- neo"Iso-, pseudo-, neo-, and epi-" indicate different types of epiisomers. isomers. Suffixes: Suffixes:  "-dine" designates isomerism as quinidine and cinchonidine. cinchonidine.  "-ine" indicates, in case of ergot alkaloids, a lower pharmacological activity e.g. ergotaminine is less potent than ergotamine. ergotamine.

Physical Properties:
I- Condition:
  

Most alkaloids are crystalline solids. solids. Few alkaloids are amorphous solids e.g. emetine. Some are liquids that are either: Volatile e.g. nicotine and coniine, or NonNon-volatile e.g. pilocarpine and hyoscine.

II- Color: IIThe majority of alkaloids are colorless but some are colored e.g.:  Colchicine and berberine are yellow.  Canadine is orange.  The salts of sanguinarine are copper-red. copper-

Physical Properties:
III- Solubility: IIIBoth alkaloidal bases and their salts are soluble in alcohol. alcohol. Generally, the bases are soluble in organic solvents and insoluble in water Exceptions: Exceptions:  Bases soluble in water: caffeine, ephedrine, codeine, water: colchicine, pilocarpine and quaternary ammonium bases. bases.  Bases insoluble or sparingly soluble in certain organic solvents: solvents: morphine in ether, theobromine and theophylline in benzene. benzene.
  

Salts are usually soluble in water and, insoluble or sparingly soluble in organic solvents. solvents. Exceptions: Exceptions:
Salts insoluble in water: quinine monosulphate. water: monosulphate. Salts soluble in organic solvents: lobeline solvents: hydrochlorides are soluble in chloroform. chloroform. and apoatropine

 

IV- Isomerization: IV     

Optically active isomers may show different physiological activities. l-ephedrine is 3.5 times more active than d-ephedrine. l-ergotamine is 3-4 times more active than d-ergotamine. 3d- Tubocurarine is more active than the corresponding lform. Quinine (l-form) is antimalarial and its d- isomer (l quinidine is antiarrythmic. The racemic (optically inactive) dl-atropine is (optically dlphysiologically active.

Chemical Properties:
I- Nitrogen:
   

Primary amines RR-NH2 Secondary amines R2-NH Tertiary amines R3-N Quaternary ammonium salts R4-N

e.g. e.g. e.g. e.g

Norephedrine Ephedrine Atropine d-Tubocurarine

II- Basicity: II R2-NH > R-NH2 > R3-N



Saturated hexacyclic amines is more basic than aromatic amines.

According to basicity Alkaloids are classified into:

  

Weak bases e.g. Caffeine Strong bases e.g. Atropine Amphoteric * Phenolic Alkaloids e.g. Morphine *Alkaloids with Carboxylic groups e.g. Narceine Neutral alkaloids e.g. Colchicine

III- Oxygen: III Most

alkaloids contain Oxygen and are solid in nature e.g. Atropine. alkaloids are free from Oxygen and are mostly liquids e.g. Nicotine, Coniine.

 Some

IV- Stability: IV Effect

of heat: heat: Alkaloids are decomposed by heat, except Strychnine and caffeine (sublimable). sublimable). with acids:

 Reaction

1- Salt formation. 2- Dil acids hydrolyze Ester Alkaloids e.g. Atropine

3- Conc. acids may cause:

 Dehydration:

Atropine Apoatropine Morphine Apomorphine  Demethoxylation: e.g. Codeine

Effect of Alkalies: 1- Dil alkalis liberate most alkaloids from their salts e.g. NH3. 2- They may cause isomerization (racemization) of alkaloid as the conversion of hyoscyamine to atropine. 3- They also can form salts with alkaloids containing a carboxylic group e.g. narceine. 4- Strong alkalis: such as aqueous NaOH and KOH form salts with phenolic alkaloids. 5- Strong alkalis cause hydrolysis of Ester alkaloids (e.g. atropine, cocaine and physostigmine) and Amide alkaloids ( colchicines). 6- Strong alkalis cause opening of lactone ring.

 Effect

of light and Oxygen:

Some alkaloids are unstable when exposed to light and Oxygen:

Oxygen

Eserine
Alkaline solutions

Rubreserine

Reserpine Ergot Alkaloids

Decomposition Lumi Alkaloids

(Inactive)

Qualitative test for Alkaloids:



Precipitation Reagents: They are used to: 1- Indicate the absence or presence of Alkaloids 2- Test for complete of extraction Disadvantages: Some non alkaloids interfere such as Proteins, lactones, coumarins

Classification of Alkaloidal precipitating agents:

a- Mayers Reagent: Potassium Mercuric Iodide. b- Dragendorffs Reagents: Potassium Iodobismethate. c- Gold Chloride.

1- Reagents that form double salts:

2- Reagents Containing Halogens:

a- Wagners Reagent: Iodine/ Potassium Iodide.

3-Organic Acids:
a- Hagers Reagent: Picric Acid b- Tannic Acid.

4- Oxygenated High Molecular Weight Acids:

a- Phosphomolybdic acid b- Phosphotungestic acid c- Silicotungestic Acid

Colour Reagents:

1- Froehds Reagent: Phosphomolybdic acid 2- Marquis Reagent: Formaldehyde/ Conc. H2SO4 3- Mandalins Reagent: Sulphovanidic acid 4- Erdmanns Reagent: Conc. HNO3/Conc. H2SO4 5- Mecke's Reagent: Selenious acid / conc. H2SO4 6- Shaer's Reagent: Hydrogen peroxide / conc. H2SO4 7- Rosenthaler's Reagent: Potassium arsenate / conc. H2SO4 8- Conc. HNO3

Extraction, Purification and Isolation of Alkaloids from Powdered plants



Extraction and purification

Method I: The powder is treated with alkalis to liberates the free bases that can then be extracted with water immiscible organic solvents. solvents. Method II: The powdered material is extracted with water or aqueous alcohol containing dilute acid. Alkaloids are extracted as acid. their salts together with accompanying soluble impurities. impurities. Method III: III: The powder is extracted with water soluble organic solvents such as MeOH or EtOH which are good solvents for both salts and free bases. bases.

Plant material and solvent

Organic solvent dissove Alkaloids Extract Organic solvent dissove Impurities

Concentration

Acidification Alkalinization

Acidified Extract (Alk. as salts) Alkaline aqueous layer

 Liberation

of the free bases:

Alkalis are used to liberate free bases. Alkalis must be strong bases. enough to liberate free bases. However, choice of strong bases. alkalis must be avoided in some cases: cases: 1- Ester Alkaloids 2- Amide Alkaloids e.g. Solanaceous Alkaloids e.g. Colchicine

3- Phenolic Alkaloids e.g. Morphine 4- Lactone Alkaloids e.g. Pilocarpine

5- Fatty Drugs due to saponification and emulsion formation.

 NH4OH:

Most widely used due to many advantages:

1- Strong enough to liberate most of alkaloids from their salts. 2- Milder than fixed alkalis so more safe. 3- Volatile so easy to get rid of it.

 Other

Alkalis:

Na2CO3, NaHCO3, Ca(OH)2, MgO.

 Extraction
 CHCl3:

of the free bases:

Strong solvent can extract most of the alkaloids. Extracts contain more impurities. Carcinogenic.
 Ether:

Gives cleaner Extract but have some disadvantages: 1- High volatility 2- Peroxide formation 3- High water miscibility

Volatile Alkaloids
 The

best way for their extraction is steam distillation. material + water + Fixed alkali Heat

 Plant

steam contain alkaloids received in acidic sloution.

Purification of the Crude Alkaloidal Fractions:

AcidAcid-Base procedures: Render extract Acidic, extract with Acidic, organic solvent (dissolve non alkaloidal impurities), Alkalinize and extract again with organic solvents (Dissolve Alkaloids). Alkaloids).  Precipitation with alkaloidal precipitating agent. agent.  Convert to crystalline salts. salts.
 Repeated

Separation of Alkaloidal Mixtures:

 Fractional

Crystallization:
oephe ri e alates

Ephe ri e & se

rystallizatio from water

Ephe ri e alate rystals

se

oephe ri e Sol tio

alate

Atropine

Hyoscyamineine Oxalates
Crystallization from Acetone/Ether

Atropine Oxalate Crystals

Hyoscyamine Oxalate olution

 Preparation

of Derivatives:
O

Separation of Primary, Secondary and Tertiary Alkaloids.

Mixture + p-toluenesulphonyl chloride
Add l a d filter

Cl

S O

Filtrate tertiary alkaloids as salt (no reaction with reagent 1ry alk derivative
O R-HN S O acidic hydrogen OH R-N S O

Precipitate 2ry alk derivative

O keto form R-N S R O insoluble in alkalis

enol form soluble in alkalis

NaOH, filter

Filtrate 1ry alk derivative

Precipitate 2ry alk derivative

Fractional Liberation:

Atropi e & yoscyami e & yosci e l salts the form of

1- Alkali ize by Na 2- Extract with Ether
3 pH

7.5

Ether Hyosci e free base (pKa = 6.2)

Aqueous layer Atropi e & Hyoscyami e HCl (pKa = 9.3)

 Fractional

Distillation: e.g. Separation of Nicotine and Anabasine

 Chromatographic

Separation.

Identification of Alkaloids:
 Melting

point  Colour test  Optical Rotation  Microcrystal test  HPLC, GC, GC-MS GC UV, IR, NMR, MS.

Quantitative Determination of Alkaloids:

 

Volumetric methods: These are based on reaction of alkaloidal bases with acids (Acid-Base titration). (AcidThey include: Aqueous titration: This is carried by either: 1- Direct titration of the alcoholic solution of the alkaloidal residue with standard acid, or 2- Back titration by dissolving the residue in a known amount of standard acid and back titration of residual acid against standard alkali. NonNon-aqueous titration: This method is suitable for determination of weak bases e.g. Caffeine.

Gravimetric methods:

These methods are recommended for determination of: of: 1- Very weak bases which can not be determined by volumetric methods e.g. caffeine and colchicine. colchicine. 2- Mixtures of alkaloids that are obtained from the same plant but differ greatly in their molecular weight e.g. Cinchona and Rawolfia alkaloids. alkaloids. They can be performed by either: either: 1- Direct Weighing of the alkaloidal mixtures 2- Precipitation of the total alkaloids and determination of the weight of the precipitate obtained. obtained. The major drawbacks of the gravimetric methods are: are: 1- They are insensitive to microamounts of alkaloids. alkaloids. 2- They could not be applied in case of thermolabile and volatile alkaloids. alkaloids. 3- Lipophilic impurities in the residue are calculated as alkaloids. alkaloids.

 Colourimetric

Method:

e.g. Morphine + NaNO2/HCl Ergot + p-dimethylaminobenzaldehyde p-

 Spectrophotometric  Polarimetric  Fluorimetric

Methods.

Method. Method. Methods

 Chromatographic

Classification of Alkaloids
 Biogenetic.
Based on the biogenetic pathway that form the alkaloids.

 Botanical

Source.

According to the plant source of alkaloids.

 Type

of Amines.

Primary, Secondary, Tertiary alkaloids.

 Basic

Chemical Skeleton

Phenylalkylamines:
e.g. Ephedrine

Pyridine and piperidine

e.g. lobeline, nicotine

Tropane
e.g. Atropine.
O

Quinoline
e.g.quinine and quinidine

Isoquinoline
e.g. papaverine

Phenantheren
e.g. Morphine

Indole
e.g.ergometrine

Imidazole
e.g. pilocarpine

Purine
e.g. caffeine
1 N 2

6 5

7 N

8 N 4 3 Puri N 9

Steroidal
e.g. Solanum and Veratrum alkaloids

Terpenoid
e.g. Taxol