ALKALOID

Lia Marliani, M.Si., Apt

Rubi Farmakognosi-Fitokimia STFB

Glucose

hn

CH2OH (6 carbons)
O
OH
HO
OH
OH

CO2
photosynthesis

CH2OH
O
OH
HO

O
OH

starch

CH2OH
O
OH

CH2OH
O
OH

O
OH

O
OH

glycolysis
CHO
CH2OH

CH OH

CHO

CH OH

HC OH

CH2OP

CH2OP

erythrose4-phosphate

C O

CH2
C OP

COOH

HO

COOH

shikimic
acid

anthanilic
acid

OH
OH

NH2

phenylpropanes

lysine
ornithine

phenylalanine
tyrosine
tryptophan

alkaloids

phosphoenol
pyruvate (PEP)

H3C C SCoA

oxaloacetate

nicotinic
acid

H3C C CH2 C CH2

lipids
fatty acids

(3 carbons)

COOH

polyketides
acetogenins

CH2OP

aspartic
acid

acetylcoenzymeA
(2 carbons)

HO

CH3

mevalonic
acid
citric
acid
cycle

energy (ATP)
+ CO2 + H2O
NH3

glutamic acid

terpenes
steroids
carotenoids

Qualitative test for Alkaloids :

a. 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

b. Colour Reagents

 Classification of Alkaloidal precipitating agents:

1- Reagents that form double salts:
a- Mayers Reagent:

Potassium Mercuric Iodide.

b- Dragendorffs Reagents: Potassium Iodobismethate.

c- Gold Chloride.

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

ALKALOIDS IDENTIFICATION

Most alkaloids are precipitated from neutral or slightly acid solution by Mayers reagent
(potassiomercuric iodide solution), by Wagners reagent (solution of iodine in potassium iodide), by
solution of tannic acid, by Hagers reagent (a saturated solution of picric acid), or by Dragendorffs
reagent (solution of potassium bismuth iodide). These precipitates may be amorphous or crystalline and
are of various colours: cream (Mayers), yellow (Hagers), reddish-brown (Wagners and Dragendorffs).
Caffeine and some other alkaloids do not give these precipitates
Caffeine, a purine derivative, does not precipitate like most alkaloids. It is usually detected by mixing
with a very small amount of potassium chlorate and a drop of hydrochloric acid, evaporating to dryness
and exposing the residue to ammonia vapour. A purple colour is produced with caffeine and other
purine derivatives

REAKSI MAYER

Reaksi Wagner

DRAGENDRORFF

POSITIF PALSU ALKALOID

Protein : mengendap dengan adanya logam berat

Tannin : mengkhelat ion logam, membentuk senyawa tak larut

Senyawa non nitrogen yang memiliki karbonil terkonjugasi atau lakton : teralkilasi mengendap. Co : kumarin,

chalchone, digitoxigenin
Pemisahan : pembasaan, diikuti, dengan ekstraksi dengan pelarut organik, kemudian diekstraksi dengan asam

encer
Isolat dari Melicope ternata (1949) bereaksi positif terhadap pereaksi alkaloid namun isolat tersebut adalah

hidroksiflavon teralkilasi :meliternatin, melliternin, dan ternatin

NEGATIF PALSU ALKALOID

Protoalkaloid : nitrogen nonheterosiklik , co : efedrin, meskalin.

Alkaloid quarterner dan amine oxide

(*contohnya ???? Kenapa ???)

Pengujian dilakukan terhadap lapisan basa dan lapisan asam

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.
Method II:
The powdered material is extracted with water or aqueous alcohol containing dilute acid.
Alkaloids are extracted as their salts together with accompanying soluble impurities.
Method 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.

EXTRACTION OF ALKALOIDS
Process A.
The powdered material is moistened with

water and mixed with lime which combines

with acids, tannins and other phenolic
substances and sets free the alkaloids (if they
exist in the plant as salts).
Extraction is then carried out with organic

solvents such as ether or petroleum spirit. The

concentrated organic liquid is then shaken with
aqueous acid and allowed to separate. Alkaloid
salts are now in the aqueous liquid, while many
impurities remain behind in the organic liquid

Process B.
The powdered material is extracted with water

or aqueous alcohol containing dilute acid.

Pigments and other unwanted materials are
removed by shaking with chloroform or other
organic solvents.
The free alkaloids are then precipitated by the

addition of excess sodium bicarbonate or

ammonia and separated by filtration or by
extraction with organic solvents

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 enough to liberate free

bases. However, choice of strong alkalis must be avoided in some cases:

1- Ester Alkaloids

e.g. Solanaceous Alkaloids

2- Amide 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 OF THE FREE BASES:

CHCl3:

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
Volatile liquid alkaloids such as nicotine and coniine are most conveniently isolated by distillation. An
aqueous extract is made alkaline with caustic soda or sodium carbonate and the alkaloid distilled off
in steam.

PURIFICATION OF THE CRUDE ALKALOIDAL FRACTIONS:

Repeated Acid-Base procedures:

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

The separation and final purification of a mixture of alkaloids may sometimes be

done by fractional precipitation or by fractional crystallization of salts such as
oxalates, tartrates or picrates. Chromatographic methods are particularly suitable
if the mixture is a complex one and if small quantities of alkaloids will suffice

Fractional Liberation:

Atropine & Hyoscyamine & Hyoscine

the form of HCl salts
1- Alkalinize by NaHCO3 pH 7.5
2- Extract with Ether

Ether
Hyoscine free base
(pKa = 6.2)

Aqueous layer
Atropine & Hyoscyamine HCl
(pKa = 9.3)

SEPARATION OF ALKALOIDAL MIXTURES:

Fractional Crystallization:

Ephedrine & Pseudoephedrine Oxalates

Crystallization from water

Ephedrine Oxalate
Crystals

Pseudoephedrine Oxalate
Solution

Atropine & Hyoscyamineine Oxalates

Crystallization from
Acetone/Ether

Atropine Oxalate
Crystals

Hyoscyamine Oxalate
Solution

PREPARATION OF DERIVATIVES:
SEPARATION OF PRIMARY, SECONDARY AND TERTIARY ALKALOIDS
O
Mixture + p-toluenesulphonyl chloride

Cl

Add HCl and filter

Filtrate
tertiary alkaloids as salt
(no reaction with reagent

Precipitate
1ry alk derivative
O
R-HN S

2ry alk derivative

O
R-N S
R
O

keto form

insoluble in alkalis
acidic hydrogen
OH
R-N S

enol form
soluble in alkalis

NaOH, filter

Filtrate

Precipitate

1ry alk derivative

2ry alk derivative

IDENTIFICATION OF ALKALOIDS:
Melting point

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

QUANTITATIVE DETERMINATION OF ALKALOIDS:

Volumetric methods:
These are based on reaction of alkaloidal bases with acids (Acid- Base titration).
They 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.

Non-aqueous titration: This method is suitable for determination of weak

bases e.g. Caffeine.

QUANTITATIVE DETERMINATION OF ALKALOIDS:

Gravimetric methods :

These methods are recommended for determination of:

1- Very weak bases which can not be determined by volumetric methods e.g. caffeine and
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.

They can be performed by either:

1- Direct Weighing of the alkaloidal mixtures
2- Precipitation of the total alkaloids and determination of the weight of the precipitate obtained.
The major drawbacks of the gravimetric methods are:
1- They are insensitive to microamounts of alkaloids.
2- They could not be applied in case of thermolabile and volatile alkaloids.
3- Lipophilic impurities in the residue are calculated as alkaloids.

QUANTITATIVE DETERMINATION OF ALKALOIDS:

Colourimetric Method:

e.g. Morphine + NaNO2/HCl

Ergot + p-dimethylaminobenzaldehyde
Spectrophotometric Methods.
Polarimetric Method.
Fluorimetric Method.

Chromatographic Methods

Correlation between Color after Spraying with Iron(III) Chloride/Perchloric Acid Spray and Chemical
Type for Mitragyna and Uncaria Heteroyohimbine Alkaloids (H & JDP)

TROPANE ALKALOID

The principal alkaloids of medicinal interest in

this group are ()-hyoscyamine; its more
stable racemate atropine, and hyoscine
(scopolamine). The compounds are esters
and are hydrolysed by heating at 60C with
baryta water; atropine yields tropic acid and
tropine; hyoscine gives tropic acid and oscine
(scopine is actually formed by enzymatic
hydrolysis but the chemical treatment
converts it to the more stable geometric
isomer, oscine).

TROPANE ALKALOID

The tropane alkaloids mainly occur in the plant family Solanaceae, but are

found as well in the families Convolvulaceae, Erythroxylaceae,

Proteaceae, and Rhizophoraceae
see Table 1 (Cordell, THE ALKALOIDS - Chemistry and

Pharmacology, volume 44, 1993)

Solanaceous Tropane Alkaloids

Occurrence:
Atropa, Datura, Hyoscyamus, Duboisia spp.

Main Alkaloids are:

1- Atropine.
2- Hyoscyamine.
3. Hyoscine (Scopolamine).

TROPANE ALKALOID STRUCTURES

3 -Monosubstituted tropanes
The 3-monosubstituted tropanes consist of 49 representatives. All
members (1-49) are formally derived from 3-hydroxytropane (1) or from
the not yet naturally found 3-hydroxynortropane.
3-Monosubstituted tropanes
The 3-monosubstituted tropanes comprise 3-hydroxytropane (SO), its five
naturally occurring ester derivatives (51,52,53, 55, and 56), and the
nortropane derivative 3benzoyloxynortropane (54).
3,6- and 3,7-disubstituted tropanes
The base compound of the large group of disubstituted tropanes (63
representatives) is 3,7-dihydroxytropane (57). All other compounds (58119) are mono- or diesters of 57 or of the corresponding 3,6-derivative

TROPANE ALKALOID STRUCTURES

3,6,7-Trisubstituted tropanes
The 3,6,7-trisubstituted tropanes (120-135) are formally derived
from the recently naturally found 3,6,7-trihydroxytropane( 120) (or
from the corresponding nortropane)
3-Substituted 6,7-epoxytropanes

The 3-substituted 6.7-epoxytropanes (136-143) are characterized by

the 6,7-epoxy ring
3-Substituted 2-carboxytropanes
The 3-substituted 2-carboxytropanes (144-150) can be considered to
be derivatives of 2-carboxy-3-hydroxytropane(ecgonine, 146) or of the
corresponding, not yet naturally found, 2-carboxy-3-hydroxynortropan

TROPANE ALKALOID STRUCTURES

3-Substituted 4-benzyltropane
The 3-substituted 4-benzyltropanes (151-155) are without
functionality at C-6 and C-7.
3,6-Disubstituted, 4-benzyltropanes

The 3-6-disubstituted 4-benzyltropanes (156-157) are 3substituted 4 benzyltropanes with an additional functionality at C-6
3,7-Disubstituted 4-benzyltropanes

The 3,7-disubstituted 4-benzyltropanes (158-160) are similar to

those of the preceding group except that the additional functionality
is at C-7 instead of C-6

TROPANE ALKALOID STRUCTURES

3,6-Disubstituted 4-benzyltropanes
The 3,6-disubstituted 4-benzyltropanes (161 and 162) are
the only 4-benzyltropanes where the C-3 substituent is

Pyranotropanes
The natural products 163-166 contain a -pyrano group
attached to the 3,4-position of the tropane ring
3,4-Dihydropyranotropanes
Compounds 167-168 are -pyranotropane derivatives in which
the 3,4-double bond is reduced

TROPANE ALKALOID STRUCTURES

l0,ll-Dihydropyranotropanes
The 10,l I-dihydropyranotropanes (169-174) are pyranotropanes in which the 10, I I -double bond is reduced. It
has not been possible to deduce from the data available
whether the C-10 and/or C-ll substituents, when present. are
or

Miscellaneous tropanes
The following heterogeneous group contains I3 "monomeric"
compounds (175-187) not falling in any of the 13 preceding
groups. Some, however, are apparent precursors for
compounds mentioned earlier [e.g., chalcostrobamine (187) for
strobamine (173

ATROPINE

()-hyoscyamine and ()-hyoscyamine (atropine) are used

medicinally. These alkaloids compete with acetylcholine for the
muscarinic site of the parasympathetic nervous system, thus
preventing the passage of nerve impulses, and are classified as
anticholinergics. Acetylcholine binds to two types of receptor
site, described as muscarinic or nicotinic.

Hyoscine

Atropa belladona Solanaceae

These Alkaloids are anticholinergic

agents:
-Decrease saliva and GIT secretions
so used preoperative.
-Decrease motility of smooth
muscles so used as antispasmodics.
-Stimulate respiratory system.
-A mydriatic (causes dilatation of
the eye pupil).
-An antidote to organophosphorus
insecticides.
-Hyoscine has more central effect so
it is sedative and hypnotic.
-Hyoscine is mainly used as
antiemetic.

Alkaloids in the form of HCl salts

1- Alkalinize by NaHCO3 pH 7.5
2- Extract with Ether

Ether
Hyoscine free base
(pKa = 6.2)

Aqueous layer
Atropine & Hyoscyamine HCl
(pKa = 9.3)
Convert to oxalate salts,
Fractional Crystallization
(Acetone/ Ether)

Atropine Oxalate
Crystals

Hyoscyamine Oxalate

Vitali-Morins test:
Solid alkaloid + fuming HNO3 Evaporate to dryness, dissolve residue in acetoVitaliMorins test:
Solid alkaloid + fuming HNO3 Evaporate to dryness, dissolve residue in
acetone, add methanolic solution of KOH Violet colour.
P-dimethylaminobenzaldehyde:
Alkaloid + reagent in porcelain dish and heat on boiling water path Intense Red Colour
Cherry Red after cooling.
Gerrards test:
Alkaloid + 2% HgCl2 in 50% Ethanol
Red colour
: Atropine
Red after warming : Hyoscyamine
White ppt
: Hyoscine

COCAINE

cocaine is of value as a local anaesthetic

for topical application. It is rapidly
absorbed by mucous membranes
and paralyses peripheral ends of sensory
nerves. This is achieved by blocking ion
channels in neural membranes
Synthetic drugs developed from the
cocaine structure have
been introduced to provide safer, less
toxic local anaesthetics

Erythroxylaceae (E.coca dan E. truxillense)

Memblok reuptake NE, serotonin, dopamin euforia,
refreshing

Memblok chanel Na anastetik

Di Amerika Selatan, digunakan dengan cara dikunyah
kapur untuk penghilang rasa lelah, lapar, dan dingin sert
meningkatkan aktivitas fisik dan ketahanan tubuh.

Cocaine

MANUFACTURE OF COCAINE
The crude alkaloids may be extracted with dilute sulphuric acid or by treatment with lime and petroleum
or other organic solvents. Non-alkaloidal matter is roughly separated by transferring the alkaloids from
one solvent to another. The crude alkaloids are obtained in solid form either as free bases by precipitation
with alkali, or as hydrochlorides by concentrating an acidified solution.
Pure cocaine is prepared from the leaves, the crude bases or the crude hydrochlorides. The process
depends on the fact that cocaine, cinnamylcocaine and -truxilline are closely related derivatives of
ecgonine, which is produced by hydrolysing them with boiling dilute hydrochloric acid
The ecgonine hydrochloride is purified and converted into the free base. This is benzoylated by interaction
with benzoic anhydride and the benzoylecgonine purified. The benzoylecgonine is methylated with methyl
iodide and sodium methoxide in methyl alcohol solution, to give methylbenzoylecgonine or cocaine. The
latter is converted into the hydrochloride and purified by recrystallization

TASK I

INDOLE ALKALOID
ISOQUINOLINE ALKALOID
PIPERIDINE ALKALOID
QUINOLIZIDINE ALKALOID

QUINOLINE ALKALOID