Hand Out Fitokimia Alkaloid 16 Des 2016

1.
Ciri umum/ kerangka utama

2. Penggolongan
3. Sifat kimia, fisika dan kromatografi
4. Ekstraksi, Fraksinasi & Pemurnian
5. Analisis fisiko kimia (UV/Vis, IR, SM, NMR)
MORPHINE - A TYPICAL ALKALOID
3 1
basic due to the contains nitrogen
unshared pair
..
Plant source. N CH3
2
Most alkaloids 4
are found in
plants. heterocyclic ring
This was the first

alkaloid discovered
(1804, Serturner).
MeO O OH
Found only in the Opium Poppy - Papaver somniferum

….. not ubiquitous.
Some Examples of Classification
BY PLANT OF ORIGIN
“Cinchona” Alkaloids
HO N
MeO quinine
“Opium” Alkaloids
N CH3
MeO O OH
morphine
BY PLANT FAMILY : “Amaryllis” Alkaloids
OH
HO
MeO belladine
O
MeO N
O
N lycorine
MeO
H
The other three
are biochemically OH
derived from MeO galanthamine
H
belladine.
H
N CH3 O N CH3
OH
O
MeO
O
tazettine
O daffodils
narcissus
These alkaloids are found in Amaryllidaceae lillies
etc
Papaver somniferum L. Coniine
Nicotiana tabacum L.
Vinblastine
Catharanthus roseus L.
Rauwolfia serpentina (L) Benth.

ALKALOID
An alkaloid is a nitrogenous
organic molecule that has a
pharmacological effect on
humans and animals.
Alkaloid :
derives from the word alkaline;

originally, the term was used to
describe any nitrogen-containing
base (an amine in modern terms).
Alkaloids
Definition: the term ―alkaloid‖ (alkali-like) is

commonly used to designate basic heterocyclic
nitrogenous compounds of plant origin that are
physiologically 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.
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.

Alkaloid bebas/ basa Garam alkaloid
HCl
••
NaOH
C18H21NO4 MW 315.364
Oxycodone Oxycodone HCl
Alkaloid bebas/ basa Garam alkaloid
- Larut dalam pelarut - Larut dalam air

organik - Jika di + basa akan
- Jika di + asam akan menjadi alkaloid bebas
menjadi garam alkaloid
New Definition:
Alkaloids are cyclic organic

compounds containing nitrogen in a
negative state of oxidation with limited
distribution among living organisms.
Nomenclature:
Trivial names should end by "ine". These names may refer to:
• The genus of the plant, such as Atropine from Atropa
belladona.
• The plant species, such as Cocaine from Erythroxylon
coca.
• The common name of the drug, such as Ergotamine from
ergot.
• The name of the discoverer, such as Pelletierine that was
discovered by Pelletier.
• The physiological action, such as Emetine that acts as
emetic, Morphine acts as narcotic.
• A prominent physical character, such as Hygrine that is
hygroscopic.
Prefixes and suffixes:
Prefixes:
• "Nor-" designates N-demethylation or N-demethoxy
lation, e.g. norpseudoephedrine and nornicotine.
• "Apo-" designates dehydration e.g. apomorphine.
• "Iso-, pseudo-, neo-, and epi-" indicate different types
of isomers.
Suffixes:
• "-dine" designates isomerism as quinidine and
cinchonidine.
• "-ine" indicates, in case of ergot alkaloids, a lower
pharmacological activity e.g. ergotaminine is less
potent than ergotamine.
Penyebaran
Penyebaran alkaloid  sangat tidak merata.
– Banyak tumbuhan yg tidak mengandungnya sama sekali
– Beberapa tumbuhan tertentu kaya akan alkaloid.
• Angiospermae : banyak ditemukan alkaloid
• Gymnospermae
• Paku-pakuan tidak atau jarang
ditemukan alkaloid
• Lumut
• Tumbuhan rendah
Penyebaran
• Tersebar luas dalam tumbuhan tinggi
• Sedikit dalam jenis jamur (Claviceps purpurea =
ergot),
• Dalam Pteridophyta hanya beberapa jenis
Lycopodium
• Dalam Gymnospermae hanya marga Ephedra
• Dari bahan bahari : mikroorganisme pada pasang
merah (―red tide‖) alkaloid neurotoksik, mis.
saksitoksin.
• Beberapa serangga memberikan feromon seks
yang bersifat alkaloid.
• Pada Monokotil, beberapa suku : Liliaceae
(paling banyak) dan Amaryllidaceae.
• Pada Dikotil paling banyak, terutama suku
Papaveraceae, Rutaceae, Leguminosae,
Rubiaceae. Terutama dalam tumbuhan daerah
iklim panas
• Papaveraceae (istimewa) : semua jenis dari
semua marga yang diteliti mengandung alkaloid.
• Labiatae dan Rosaceae hampir bebas dari
alkaloid.
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
• Alkaloid: metabolit sekunder yang mengandung nitrogen (N)
(diturunkan dari asam amino : Phe, Tyr, Trp, Lys, dan Orn)
• Tersebar pada ~ 20% tumbuhan tingkat tinggi

Alkaloid tropan: Solanaceae, Proteaceae, Euphorbiaceae,
Rhizophoraceae, Convolvulaceae, dan Cruciferae
Alkaloid benzilisokuinolin: Lauraceae, Magnoliaceae, Annonaceae
Alkaloid kuinolizidin: Leguminosae
Golongan senyawa bahan alam sd 1980 an
Terdapat
Golongan Jumlah
dalam Keterangan
senyawa senyawa
tanaman
Setiap tanaman
Hampir memiliki jenis terpenoid
Terpenoid 30.000
semua sama dan ada juga
yang berbeda
Setiap tanaman
20 %
Alkaloid 12.000 memiliki beberapa jenis
tumbuhan
alkaloid yang berbeda
Banyak tanaman
Hampir
Flavonoid 2.500 memiliki jenis flavonoid
semua
yang sama
Function in Plants
• They may act as protective against insects
and herbivores due to their bitterness and
toxicity.
• They are, in certain cases, the final products
of detoxification (waste products).
• Source of nitrogen in case of nitrogen
deficiency.
• They, sometimes, act as growth regulators in
certain metabolic systems.
• They may be utilized as a source of energy in
case of deficiency in carbon dioxide
assimilation.
THE PURPOSE OF ALKALOIDS IN PLANTS (?)
The spectacular pharmacological properties of many of the
alkaloids keeps asking about their purpose in plants.
Many ideas have been advanced:

1. Defense Mechanisms
2. Growth Regulation
3. Herbivore Attractants
4. Nitrogen Storage
5. Anti-fungals
6. Metal ion transport (chelates)
7. Competitive Herbicides
8. Insect Attractants & Repellants
What seems most likely is that there are many reasons why plants
elaborate alkaloids, and in many cases the purpose of the alkaloid
may be unique to a given plant.
Fungsi bagi manusia
Banyak senyawa alkaloid yang punya
aktivitas biologi dan telah digunakan
secara luas dalam pengobatan
1. Kinin  antimalaria
2. Morfin  analgetika – narkotika
3. Kodein  antitusif
4. Reserpin  obat jantung
5. Kokain  anestetika lokal
Glucose
hn CH2OH (6 carbons) CH2OH CH2OH CH2OH
O O O O
OH OH OH OH
HO OH HO O O O
CO2 OH OH OH OH
photosynthesis starch n
glycolysis
CHO
CH OH CHO CH2OH
HC OH C O
CH OH polyketides
CH2OP
CH2OP erythrose- CH2OP
acetogenins
4-phosphate CH2
phosphoenol O O lipids
C OP
pyruvate (PEP) H3C C CH2 C CH2 fatty acids
COOH
COOH shikimic (3 carbons)
acid anthanilic
acid
HO OH O
acetyl-
COOH
OH H3C C SCoA
coenzymeA
(2 carbons)
NH2 HO CH3
mevalonic
phenylpropanes
lysine oxalo- acid
O
phenylalanine ornithine acetate citric O
energy (ATP)
tyrosine acid + CO2 + H2O
tryptophan cycle
terpenes
NH3 steroids
nicotinic aspartic
alkaloids acid acid carotenoids
glutamic acid
Berdasarkan asal asam amino
• Turunan ornitina  atropina

• Turunan lisina  piperidina
• Turunan tirosin / fenilalanin  efedrin
• Turunan triptofan  kinin
• Turunan histidin  histamin
• Turunan asam antranilat  aktinomisin D
• Turunan asam nikotinat  arekolina
MOST ALKALOIDS ARE DERIVED FROM a-AMINO ACIDS
SOME OF THE MAJOR RELATIONSHIPS ARE SHOWN BELOW
COOH
COOH R
N NH2
NH2 NH2
H
ornithine
N N
H H
COOH tryptophan
NH2 N
NH2
lysine H
R R
and N
COOH R
NH2 NH2
R= H phenylalanine and
N
R
R = OH tyrosine R = H, CH3
R'
R‘ = H, alkyl
Ex: FORMATION OF ISOQUINOLINE ALKALOIDS
decarboxylation hydroxylation
COOH
1 2
HO
NH2 NH2 NH2

HO HO HO ..
O
Mannich reaction and 3
aromatic substitution H C CH3
H + ..
HO 5 O 4 H O
..
N H +
HO HO
H
..N H HO H N H
CH3
H CH3 CH3
Enz-B:
imine formation
methylation 6,7,8
CH3 O
Methylations could also N CH3

have taken place before CH3 O
Acetaldehyde CH3CHO
step three. CH3 is a readily available
metabolic intermediate
1. Ciri umum/ kerangka utama
2. Penggolongan
MORPHINE - A TYPICAL ALKALOID
basic due to the contains nitrogen

unshared pair
..
Plant source. N CH3
Most alkaloids
are found in
plants. heterocyclic ring
This was the first

alkaloid discovered
(1804, Serturner).
MeO O OH
Found only in the Opium Poppy - Papaver somniferum

….. not ubiquitous.
DETEKSI
Prosedur deteksi alkaloid pada sampel tumbuhan
Bahan Tumbuhan • Daun segar ~5-10 lembar
(digerus halus dengan pasir bersih)
• ekstraksi dengan • Serbuk kering kulit/kayu batang ~5-10 g
CHCl3/NH3
• saring
Residu Ekstrak CHCl3/NH3
• partisi dengan lar. 5% H2SO4
Lapisan air Lapisan CHCl3
• Uji Dragendorf
• Uji Meyer
• Uji Wagner
DETEKSI
Pereaksi-pereaksi untuk mendeteksi alkaloid:
Pereaksi tetes:
• Pereaksi Dragendorf: larutan senyawa
kompleks Bi(NO3)3/KI dalam asam
Positif  endapan jingga kemerahan
• Pereaksi Meyer: larutan senyawa
kompleks HgCl2/KI
Positif  endapan putih
• Pereaksi Wagner: larutan senyawa
kompleks KI/I2
Positif  endapan coklat
endapan jingga kemerahan
endapan putih
General tests for alkaloids:
1) Alkaloidal precipitating reagents:
A) Mayer‘s reagent. B) Wagner‘s reagent.
C) Hagar‘s reagent D) Gold chloride
E) Dragendorff‘s reagent.
2) Alkaloidal coloring reagents:

(The rough dehydration or oxidation of the alks)
A) Marquis reagent. B) Mandalin R.
C) Froehed‘s reagent D) Erdmann‘s R.
3) Special tests for alkaloids:

Used to identify or to confirm the identity of the alks.
Physical Properties:
I- Condition:
• Most alkaloids are crystalline solids.
• Few alkaloids are amorphous solids e.g. emetine.
• Some are liquids that are either:
Volatile e.g. nicotine and coniine, or
Non-volatile e.g. pilocarpine and
hyoscine.
II- Color:
The 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.
Physical Properties:
III- Solubility:
• Both alkaloidal bases and their salts are soluble in
alcohol.
• Generally, the bases are soluble in organic solvents
and insoluble in water
Exceptions:
• Bases soluble in water: caffeine, ephedrine, codeine,
colchicine, pilocarpine and quaternary ammonium bases.
• Bases insoluble or sparingly soluble in certain organic
solvents: morphine in ether, theobromine and theophylline
in benzene.
• Salts are usually soluble in water and, insoluble or

sparingly soluble in organic solvents.
Exceptions:
• Salts insoluble in water: quinine monosulphate.
• Salts soluble in organic solvents: lobeline and apoatropine
hydrochlorides are soluble in chloroform.
IV- Isomerization:
• 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.
• d- Tubocurarine is more active than the
corresponding l- form.
• Quinine (l-form) is antimalarial and its d- isomer
quinidine is antiarrythmic.
• The racemic (optically inactive) dl-atropine is
physiologically active.
Chemical Properties:
I- Nitrogen:
• Primary amines R-NH2 e.g. Norephedrine
• Secondary amines R2-NH e.g. Ephedrine
• Tertiary amines R3-N e.g. Atropine
• Quaternary ammonium salts R4-N e.g d-Tubocurarine
II- Basicity:
• 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:
• Most alkaloids contain Oxygen and are

solid in nature e.g. Atropine.
• Some alkaloids are free from Oxygen and

are mostly liquids e.g. Nicotine, Coniine.
IV- Stability:
• Effect of heat:
Alkaloids are decomposed by heat, except
Strychnine and caffeine (sublimable).
• Reaction with acids:

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 Rubreserine
Alkaline solutions
Reserpine Decomposition
Ergot Alkaloids Lumi Alkaloids

(Inactive)
BAGAN EKSTRAKSI ALKALOID
(G.A. CORDELL)
Bahan tumbuhan
eter minyak bumi
Ekstraksi eter minyak Ampas

bumi i) MeOH atau EtOH 95%
ii) Pemekatan
iii) Partisi EtOAc – asam
tartrat 2%
Ekstrak EtOAc Lapisan asam tartrat 2%

(alkaloid basa lemah Dibasakan dengan
atau netral) NH4OH atau Na2CO3
Partisi dgn EtOAc
Ekstrak EtOAc Lapisan air basa

(Fraksi alkaloid basa) (Fraksi air basa)
Diekstraksi
dgn CHCl3
Ekstraksi CHCl3 Lapisan air

(Alkaloid kuarterner)
Golongan senyawa bahan alam
Golongan
Kepolaran Isolasi Analisis
senyawa
Umumnya Relatif Relatif paling sulit
Terpenoid
non-polar mudah (MS : GCMS/LCMS)
Ada yang Mudah
Alkaloid polar dan (dengan Relatif sulit
non-polar ECC)
Umumnya Relatif Paling mudah
Flavonoid
polar sulit (Spektro UV/VIS)
CLASSIFICATION AND
STRUCTURAL TYPES
OF ALKALOIDS
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

Klasifikasi alkaloid menurut Cordell
(berdasarkan asal biosintesis)
Asal
Jenis alkaloid Posisi N
biosintesis
True alkaloid
(alkaloid sejati) Asam amino Hetero siklik
Proto alkaloid/
amino alkaloid Asam amino Alifatik
Bukan asam
Pseudo alkaloid Hetero siklik
amino
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.
Klasifikasi alkaloid
Menurut Cordell N N N N
H H H
• True Alkaloid pyrrolidine pyrrole piperidine pyridine
Alkaloid sejati
• Protoalkaloid
• Pseudoalkaloid N N N
N
H H
quinoline isoquinoline indole dihydroindole
Pseudo-alkaloid
C6 H5 CH3 H3CO NH2
O
Solasodin C C H H3CO
H3C
N
N H
OH HNCH3 OCH3
O N N
CH3 CH3 Proto-alkaloid
Efedrin Meskalin
caffeine
True alkaloids
True alkaloids derive from amino acid and they

share a heterocyclic ring with nitrogen.
These alkaloids are highly reactive substances with

biological activity even in low doses.
All true alkaloids have a bitter taste and appear as

a white solid, with the exception of nicotine
which has a brown liquid.
True alkaloids
True alkaloids form water-soluble salts.
Moreover, most of them are well-defined

crystalline substances which unite with acids to
form salts.
True alkaloids may occur in plants

(1) in the free state,
(2) as salts and
(3) as N-oxides.
True Alkaloid
• Umumnya toksik
• Mempunyai berbagai aktivitas fisiologi
• Semua bersifat basa,
• Memiliki atom N dalam cincin heterosiklik,
• Berasal dari asam amino yang dikondensasi
dengan satuan struktur non nitrogen,
• Terdapat dalam suku tertentu, pada
tumbuhan, umumnya sebagai garam dengan
asam organik.
True Alkaloid
N N N N
H H H
pyrrolidine pyrrole piperidine pyridine
N N N
N
H H
Protoalkaloids
Protoalkaloids are compounds, in which

the N atom derived from an amino acid
is not a part of the heterocyclic.
Such kinds of alkaloid include compounds

derived from l-tyrosine and l-tryptophan
Protoalkaloid
• Bersifat basa.
• Amina relatif sederhana, sering disebut sbg amina
biologi (―biological amines‖).
• Hasil dekarboksilasi asam amino
• Atom N dari asam aminonya tidak terletak pada cincin
heterosiklik.
Contoh :
C6 H5 CH3 H3CO NH2
C C H H3CO
H
OH HNCH3 OCH3
Efedrin Meskalin
Amine alkaloids
Alkaloids without heterocyclic nitrogen atoms

Pseudoalkaloids
Pseudoalkaloids are compounds, the basic carbon

skeletons of which are not derived from amino
acids.
In reality, pseudoalkaloids are connected with amino

acid pathways.
They are derived from the precursors or postcursors

(derivatives the indegradation process) of amino
acids. They can also result from the amination and
transamination reactions of the different pathways
connected with precursors or postcursors of amino
acids.
Pseudoalkaloids
These alkaloids can also be derived from

non-aminoacid precursors.
The N atom is inserted into the molecule at a

relatively late stage, for example, in the case of
steroidal or terpenoid skeletons.
Pseudoalkaloid
Tidak berasal dari prazat
asam amino,
umumnya bersifat
basa.
Terdiri dari 2 golongan :
Solasodin
1. Alkaloid steroid
(tomatidin, solasodin
O
dll) H3C N
N
2. Alkaloid purin O N N
CH3 CH3
(kafein, teobromin,
teofilin). caffeine
Steroidal alkaloids
Steroidal alkaloids
Diterpenoid alkaloids
HOW ARE ALKALOIDS CLASSIFIED ?
Common classification schemes :
(1) Biosynthetic Origin:
Depends on types of precursors or building block

compounds used by living organisms to synthesize
complex structures.
metabolic pathway
Berdasarkan asal asam amino
• Turunan ornitina  atropina

• Turunan lisina  piperidina
• Turunan tirosin / fenilalanin  efedrin
• Turunan triptofan  kinin
• Turunan histidin  histamin
• Turunan asam antranilat  aktinomisin D
• Turunan asam nikotinat  arekolina
MOST ALKALOIDS ARE DERIVED FROM a-AMINO ACIDS
SOME OF THE MAJOR RELATIONSHIPS ARE SHOWN BELOW
COOH
COOH R
N NH2
NH2 NH2
H
ornithine
N N
H H
COOH tryptophan
NH2 N
NH2
lysine H
R R
and N
COOH R
NH2 NH2
R= H phenylalanine and
N
R
R = OH tyrosine R = H, CH3
R'
R‘ = H, alkyl
Ex: FORMATION OF ISOQUINOLINE ALKALOIDS
decarboxylation hydroxylation
COOH
1 2
HO
NH2 NH2 NH2

HO HO HO ..
O
Mannich reaction and 3
aromatic substitution H C CH3
H + ..
HO 5 O 4 H O
..
N H +
HO HO
H
..N H HO H N H
CH3
H CH3 CH3
Enz-B:
imine formation
methylation 6,7,8
CH3 O
Methylations could also N CH3

have taken place before CH3 O
Acetaldehyde CH3CHO
step three. CH3 is a readily available
metabolic intermediate
Common classification schemes :
(2) Taxonomic:
Accord to the source of compound without reference to the
chemical types.
Vinca alkaloids
The Vinca alkaloids are a subset of drugs that are
derived from the periwinkle plant, Catharanthus
roseus. OH
N C2H5
N
H3COOC
N
H
MeO N OAc
H
R HO COOCH3
Vinblastine R=-CH3
Vincristine R=-CHO
Aconite alkaloids
The aconite alkaloids are derived from the aconitum

plant. They are also diterpenoids.
Berdasarkan asal tumbuhannya
• Alkaloid Solanaceae  solanin, atropina
• Alkaloid Loganiaceae  Kurare, strikhnin
• Alkaloid Papaveracea  papaverin, morfin,
kodein
• Alkaloid Rubiaceae  kinin, kinidin,
sinkonin
• Alkaloid Leguminosae  fisostigmin
ALKALOIDS
Ephedra
Datura
Opium poppy
BY PLANT OF ORIGIN
HO N
MeO quinine
N CH3
MeO O OH
morphine
OH
HO
MeO belladine
O
MeO N
O
N lycorine
MeO
H
The other three
H
belladine.
H
N CH3 O N CH3
OH
O
MeO
O
tazettine
O daffodils
narcissus
etc
(3) Chemical:
Depends on the type of heterocyclic ring systems

present.
It is usual to classify alkaloids according to the

chemical structure associated with the amino
acids (or their derivatives) from which they arise.
Berdasarkan strukturnya
1. Tropan  atropina
2. Pirolizidin  indisin -N- oksida
3. Piperidin  N-metil pelletierin, piperin
4. Kuinolizidin  lupinin, spartein
5. Indolizidin  kastanospermin
6. Piridin  nikotin, risinin
7. Feniletilamin  meskalin, anhalamin
8. Benzil tetrahidro isokinolin  tubokurarin
9. Fenil etil isokinolin  autumnalin, kolkhisin
10. Tetrahidro isokinolin  emetin, sefaelin
11. Indol  psilosin, psilosibin
12. Indol terpenoid  yohimbin, reserpin
13. Kuinolin  kinin, kinidin, kamtotesin
14. Piroloindol  fisostigmin, kimonantin
15. Ergot  ergin, ergometrin
16. Kuinazolin  peganin
17. Akridin  akrosinin
18. Imidazol  histamin, pilokarpin
19. Purin  kafein, teofilin, teobromin
HETEROCYCLIC RING SYSTEMS
LEARN THESE RINGS (plus the ones on the next page)
N N N N
H H H
N N N
N
H H
HETEROCYCLIC RING SYSTEMS (cont)
LEARN THESE ALSO
H
N
N N
quinolizidine pyrrolizidine tropane
N N
N
C C N
N N
H
benzylisoquinoline purine -phenylethylamine

BY RING TYPE
MeO O
OMe -O P OH H3C CH3
NH OMe N
MeO O + CH3
N psilocybin
N
emetine H
O
H3C N
N
N N
O N
H
CH3 CH3
N CH3
nicotine caffeine
 Phenylalkylamines:
e.g. Ephedrine CH2 CH CH3
NH 2
 Pyridine and piperidine

e.g. lobeline, nicotine
N N
H
 Tropane
e.g. Atropine. NCH3 OH
 Quinoline
e.g.quinine and quinidine
 Isoquinoline
e.g. papaverine
N
 Phenantheren
e.g. Morphine
 Indole
e.g.ergometrine
N
H
 Imidazole N
e.g. pilocarpine
 Purine 6 7
H
5 N
e.g. caffeine 1 N
8
2
N 4 N
9
3
Purine
 Steroidal
e.g. Solanum and Veratrum
alkaloids
 Terpenoid
e.g. Taxol
Tropane alkaloids
Many of these psychoactive alkaloids

Pyrrolizidine alkaloids
Piperidine alkaloids
Piperidine/Pyridine alkaloids
Purine alkaloids
Figure 3-5 Caffeine

Quinoline alkaloids
Isoquinoline alkaloids
Indole alkaloids
Vindoline
Vinblastine (Catharanthus)
Quinolizidine alkaloids
Steroidal alkaloids
Steroidal alkaloids
Diterpenoid alkaloids
Amine alkaloids
Alkaloids without heterocyclic nitrogen atoms

Other classes of alkaloids
• Betalain alkaloids - pigment alkaloids

• Lycopodium alkaloids - restricted to club
mosses
• Peptide alkaloids
1. Tropan
Alkaloid tropan merupakan kelompok besar.
Hiosiamin dan hioscin adalah termasuk
alkaloid alami yang paling penting digunakan
untuk pengobatan
Umumnya ditemukan pada Solanaceae :

• Atropa belladona
• Duboisia myoporoides, Duboisia leichhardtii
• Datura stramonium, Datura sanguinea
• Hyocyamus niger, H. muticus
• Scopollia carniolica
• Mandragora officinarum
Tropane alkaloids
• a 8-methyl-8-aza-bicyclo[1,2,3]octane or tropane
skeleton
• without the 8-methyl group, is known as nortropane.
• atropine, cocaine and scopolamine.
• occur mainly in plants from the families Solanaceae
and Erythroxylaceae (古柯科).
Tropane alkaloids (cont.)
Atropine (阿托品)
•C17H23NO3
•the first tropane alkaloid isolated from the ‗deadly
nightshade‘ (Atropa belladonna, 颠茄)
•also found in many other plants of the Solanaceae.
•a racemic mixture of D-hyoscyamine and L-hyoscyamine.
Atropine (阿托品)
• a racemic(外消旋的) mixture of D-hyoscyamine and L-hyoscyamine.

• However, most of the pharmacological properties of atropine are due
to its L-isomer,
• a competitive antagonist of the muscarinic acetylcholine receptors
due to its binding to muscarinic acetylcholine receptors.
• The main medicinal use of atropine is as an opthalmic drug.
• Usually a salt of atropine, e.g. atropine sulphate, is used in
pharmaceutical preparations .
• It is also used as an antidote for poisoning by organophosphate
insecticides and nerve gases.
major adverse effects ：

ventricular fibrillation (室颤)
tachycardia ( 心动过快)
nausea (恶心)
blurred vision (视力模糊)
loss of balance
photophobia (畏光).
overdoses ofatropine can be fatal.
the antidote of atropine poisoning is physostigmine or pilocarpine.
C17H21NO4
a white crystalline tropane alkaloid found mainly in coca plant.
a potent CNS stimulant and appetite suppressant.
For its euphoretic (精神愉快的)effect, cocaine is often used

recreationally, and it is one of the most common drugs of abuse and
addiction.
also used as a topical anaesthetic in eye, throat and nose surgery.

possession, cultivation and distribution of cocaine is illegal for non-medicinal
and non-government sanctioned purposes virtually all over the world.
side-effects of cocaine
twitching(颤搐) paranoia(狂想) impotence(阳萎)
excessive dosage
hallucinations(幻觉) paranoid delusions (狂想狂)
tachycardia, itching(搔痒) formication(蚁走感)
overdose
tachyarrhythmias(快速性心律失常)
elevated blood pressure
fatal
Hyocyamus niger Scopolia carniolica
Datura stramonium Atropa belladona
• Hiosiamin : senyawa optik aktif, (-)hiosimain
dan (+)hiosiamin
• Enantiomer (+) 20 – 30 x lebih aktif dari
bentuk (-)
• Bentuk rasemik dari kedua hiosiamin 
atropin.
• Hiosiamin dan atropin berkompetisi
dengan asetilkolin  antikolinergik.
• Atropin = antidot keracunan inhibitor
kolinesterase (fisostigmin dan neostigmin)
dan insektisida organofosfat.
Tropan (lanjutan)
2. Kokain jenis alkaloid tropan langka dan hanya ditemukan
dari suku Erythroxylaceae, (Erythroxylon coca dan E.
truxillense.)
• Kokain digunakan untuk pengobatan sebagai anestetik
lokal dan penyegar karena sifat euforianya.
• Daun koka digunakan dengan cara dikunyah kapur di
Amerika Selatan untuk :
– obat penghilang rasa lelah, lapar dan dingin
– meningkatkan aktivitas fisik dan ketahanan tubuh.
• Daun koka : 0,7 – 2,5 % alkaloid (kokain 40 – 50 %,
kuskohigrin 20 – 30%, sinnamoilkokain, tropakokain).
• Mulai 1800, Coca cola dikenal sebagai minuman
penyegar yg mengandung kokain dan kafein
• Sejak tahun 1906 tidak diperbolehkan dalam minuman
Drugs - Cocaine
Mechanism of Action
– Blocks reuptake of NE, Serotonin, Dopamine
– Blocks fast sodium channels – anesthetic effect
$40-50 per ¼ gram (1 use);
$80-100/ 1g
2. Quinoline alkaloids
Any alkaloid that possesses a quinoline, i.e. 1-
azanaphthalene, 1-benzazine, or benzo[b]pyridine,
skeleton is known as a quinoline alkaloid,
e.g. quinine. Quinoline itself is a colourless hygroscopic
liquid with strongodour, and slightly soluble in water, but
readily miscible with organicsolvents. Quinoline is toxic.
Short term exposure to the vapour of quinoline causes
irritation of the nose, eyes, and throat, dizziness and
nausea. It may also cause liver damage.
Quinine, molecular formula C20H24N2O2, is a white crystalline
quinoline alkaloid, isolated from Cinchona bark (Cinchona succirubra), and is
well known as an antimalarial drug. Quinine is extremely bitter, and also
possesses antipyretic, analgesic and anti-inflammatory properties. While
quinine is still the drug of choice for the treatment of Falciparum malaria, it
can be also used to treat nocturnal leg cramps and arthritis. Quinine is an
extremely basic compound, and is available in its salt forms, e.g. sulphate,
hydrochloride and gluconate.
Despite being a wonder drug against malaria, quinine in therapeutic doses
can cause various side-effects, e.g. nausea, vomiting and cinchonism, and in
some patients pulmonary oedema. It may also cause paralysis if accidentally
injected into a nerve. An overdose of quinine may have fatal consequences.
Non-medicinal uses of quinine include its uses as a flavouring agent in tonic
water and bitter lemon.
Quinidine, molecular formula C20H24N2O2,
is a stereoisomer of quinine found in
Cinchona bark. Chemically, it is known as (2-
ethenyl-4-azabicyclo [2.2.2] oct-5-yl)-(6-
methoxyquinolin-4-yl)-methanol, or 60-
methoxycinchonan-9-ol. It is used as a Class
1 anti-arrhythmic agent.
Intravenous injection of quinidine is also used

in the treatment of P. falciparum malaria.
Among the adverse effects, quinidine induces
thrombocytopenia (low platelet counts) and
may lead to thrombocytic purpurea.
2. Kuinolin
• Alkaloid kina Cinchona sp (Rubiaceae),
C. succirubra, C. ledgeriana, dan C. calisaya.
dipanen setelah 8-12 thn.
• Alkaloid kina utama : kinin, kinidin, sinkonin dan
sinkonidin.
• Kinin untuk pengobatan malaria Plasmodium
falciparum dengan dosis 600 mg 3x sehari.
• Kinin untuk tonikum pada dosis 80 mg/L
sekaligus untuk mencegah malaria.
• Kinin juga untuk relaksan otot, &mengobati kram.
• Kinidin selain untuk malaria, juga untuk
aritmia jantung
• Kinidin juga ditemukan pada kulit batang

Remija pendunculata (Rubiaceae).
• Sinkonidin, banyak dicari karena
harganya 10 x lebih mahal dari kinin.
Digunakan untuk katalis pada reaksi
stereokimia.
ARAH KEBIJAKAN PENGEMBANGAN
KINA NASIONAL
• Kebutuhan kina dunia 10.000 ton per tahun, baru

dapat dipenuhi 7.700 ton per tahun
• Kebutuhan kina Indonesia per tahun 3.000 ton –
5.000 ton pertahun, 80% di impor dari Afrika
• Luas lahan kina Indonesia 9.900 ha, 5.000 ha lahan
rakyat, 4.900 ha perkebunan besar swasta/negara.
• Produksi per tahun 1.500 – 1.800 ton kulit kering
• Produksi kina Afrika mulai menurun
• Kina sebagai obat malaria tersaingi artemisinin
Alkaloid Content Comparison by Cinchona
species
Species Total Alkaloids Quinine Content
(%) (%)
C. calisaya 3-7 0-4
C. pubescens 4.5 - 8.5 1-3
C. officinalis 5-8 2 - 7.5
C. ledgeriana 5 -14 3 - 13
C. succirubra 6 - 16 4 - 14
3. Isoquinoline alkaloids
Isoquinoline is in fact an isomer of quinoline, and chemically
known as benzo[c]pyridine or 2-benzanine. Any alkaloids that
possess an isoquinoline skeleton are known as isoquinoline
alkaloids, e.g. papaverine and morphine.
The isoquinoline backbone is biosynthesized from the
aromatic amino acid tyrosine.
Isoquinoline itself is a colourless hygroscopic liquid at
room temperature. It has an unpleasant odour. It is slightly
soluble in water but well soluble in ethanol, acetone, ether
and other common organic solvents. Isoquinoline is a
weak base with a pKa of 8.6. Isoquinoline alkaloids play an
important part in medicine. A number of these alkaloids are
available as drugs. Some examples of isoquinoline
derivatives with medicinal values are summarized in the
following table. In addition to their medicinal uses,
isoquinolines are used in the manufacture of dyes, paints
and insecticides, and as a solvent for the extraction of
resins.
Papaverine, molecular formula C20H21NO4, is an isoquinoline
alkaloid isolated from poppy seeds (Papaver somniferum, family
Papaveraceae). This alkaloid is used mainly in the treatment of spasms
and of erectile dysfunction. It is also used as a cerebral and coronary
vasodilator. Papaverine may be used as a smooth muscle relaxant in
microsurgery. In pharmaceutical preparations, papaverine is used in its
salt form, e.g. hydrochloride, codecarboxylate, adenylate and teprosylate.
The usual side-effects of papaverine treatment include polymorphic
ventricular tachycardia, constipation, increased transaminase levels,
hyperbilirunemia and vertigo.
Morphine (C17H19NO3), a habit forming Class A analgesic drug,
is the major bioactive constituent of opium poppy seeds. Like other
opium constituents (opiates), e.g. heroin, morphine acts directly on the
CNS to relieve pain. Morphine is used for the treatment of post-surgical
pain and chronic pain (e.g. cancer pain), and as an adjunct to general
anaesthesia, and an antitussive for severe cough. Side-effects of
morphine treatment generally include impairment of mental performance,
euphoria, drowsiness, loss of appetite, constipation, lethargy and blurred
vision.
3. Isoquinolin
• Opium adalah eksudat atau lateks dari kapsul
yang belum masak dari Papaver somniferum
(Papaveraceae).
Kapsul ditunggu sampai masak (berubah warna

dari biru-hijau  kuning), disayat untuk
mengeluarkan lateks.
• Opium untuk analgetik dan narkotik dan obat

batuk.
Kandungan opium standar adalah 12 %.
• Morfin adalah analgetik dan narkotik kuat,
penghilang rasa sakit yang parah.
Efek sampingnya : penurunan mental,
ketagihan, mual, muntah, diare dll.
• Kodein : analgetik yang aman dan tidak
menimbulkan ketagihan, juga sbg obat batuk.
• Tebain digunakan sbg antagonis morfin.

• Papaverin adalah alkaloid benzilisokuinolin yg
ditemukan pada opium
• Papaverin : untuk ekspektoran dan impoten.
Indole alkaloids
Indole chemistry has already been discussed in Chapter 4.
This is one of the major groups of naturally occurring
bioactive alkaloids, and can be classified into three main
categories: tryptamine and its derivatives, ergoline
and its derivatives, and b-carboline and its derivatives.
Tryptamine derivatives Tryptamine, chemically known as 3-(2-
aminoethyl)indole, is widespread in plants, fungi and animals. Biosynthetically,
tryptamine derives from the amino acid tryptophan. Tryptamine acts
as the precursor of many other indole alkaloids. Substitutions to the
tryptamine skeleton give rise to a group of compounds collectively known
as tryptamines: e.g. serotonin, an important neurotransmitter, is the 5-
hydroxy derivative of tryptamine; melatonin, a hormone found in all living
creatures, is 5-methoxy-N-acetyltryptamine. Some of the pharmacologically
active natural tryptamines are psilocybin (4-phosphoryloxy-N,N-dimethyltryptamine)
from ‗magic mushrooms‘ (Psilocybe cubensis and P. semilanceata),
DMT (N,N-dimethyltryptamine) from a number of plants and DET
(N,N-diethyltryptamine), an orally active hallucinogenic drug and psychedelic
compound of moderate duration. Many synthetic tryptamines, e.g.
sumatripan (5-methylaminosulphonyl-N,N-dimethyltryptamine), a drug
used for the treatment of migraine, are also available.
4. Indol
1. Reserpin dan deserpidin adalah kandungan
aktif dari Rauwolfia serpentina, R. canescens,
R. vomitoria (Apocynaceae).
• Secara klinis terbukti sbg antihipertensi dan
sedatif.
• Rauwolfia sudah 3000 th sbg obat tradisional
untuk anti bisa ular, mengobati sakit perut,
demam, muntah dan sakit kepala.
• Kandungan total alkaloid Rauwolfia 0,7-2,4%
tetapi yang aktif hanya 0,15-0,2% yaitu
reserpin, deserpidin dan resinamin.
• Reserpin dan deserpidin sbg obat
antihipertensi dan transquiliser dengan
mengintervensi penyimpanan katekolamin.
Pemakaian dalam waktu lama
menyebabkan depresi berat.
• Reserpin juga untuk obat kanker
payudara.
• Ajmalisin secara klinis untuk antihipertensi
• Ajmalin untuk aritmia jantung.
• Pemisahan sesama alkaloid Rauwolfia
berdasarkan kebasaannya :
– serpentin paling kuat sifat basanya,

– ajmalin dan senyawa sejenisnya
kebasaannya menengah
– reserpin, resinamin, deserpidin dan
ajmalisin merupakan basa lemah
Reserpin
Rauwolfia serpentina
3. Indol (lanjutan)
2. Alkaloid indol dari tanaman Chataranthus roseus
(=Vinca rosea) menghasilkan beberapa indol alkaloid
seperti vinblastin (VBL), vinkristin (VCR), vindolin (V),
vinleurosin, vinrosidin dan leurokristin (LC).
• Vinblastin dan vinkristin adalah obat anti kanker yang
penting.
• Vinblastin terutama untuk mengobati penyakit Hodgkin‘s
yaitu kanker akibat kelenjar limfa, pankreas dan hati.
• Vinkristin lebih aktif dibanding vinblastin tetapi lebih
neurotoksik dan terutama digunakan untuk leukemia
pada anak, juga untuk limfoma, kanker paru-paru,
serviks dan payudara.
• Masalah utama pada penggunaan VBL
dan VCR adalah kadar dalam tanaman
yang sangat kecil (0,0002%)  dari 500
kg tanaman diperoleh 1 g VCR.
• Dalam tanaman, VBL lebih tinggi kadarnya

dibanding VCR. Dengan bantuan
Streptomyces albogriseolus, VBL dapat
diubah menjadi VCR.
Ergolines Alkaloids that contain an ergoline skeleton are called
ergolinealkaloids, and some of them are psychedelic drugs, e.g. LSD. A
number ofergoline derivatives are used clinically as vasoconstrictors (e.g. 5-
HT 1agonists, ergotamine), and in the treatment of migraine and
Parkinson‘sdisease, and some are implicated in the disease ergotism.Ergine,
molecular formula C16H17N3O, is the amide of D-lysergic acid,and
commonly known as LSA or LA-111. It is an ergoline alkaloid that occurs in
various species of the Convolvulaceae, and in some species of fungus.
Rivea corymbosa (ololiuqui), Argyreia nervosa (Hawaiian baby woodrose)
and Ipomoea violacea (tlitliltzin) are three major sources of this
alkaloid.
Lysergic acid diethylamide, molecular formula C20H25N3O,
also known as LSD or LSD-25, is a semi-synthetic
psychedelic drug, synthesized from the natural precursor
lysergic acid found in ergot, a grain fungus. It is a
colourless, odourless and mildly bitter compound. LSD
produces altered experience of senses, emotions,
memories, time and awareness for 8 to 14 h.
Moreover, LSD may cause visual effects, e.g. moving

geometric patterns, ‗trails‘ behind moving objects and
brilliant colours.
5. Ergot
• Ergot adalah jamur Claviceps purpurea yang
menyerang rumput-rumputan Triticum
aestivum dan Secale cereale (Graminae).
• Alkaloid ergot dapat meracuni hewan dan
manusia dengan gejala :
– Gangguan saluran cerna : diare, muntah, sakit
perut
– Gangguan peredaran darah : kedinginan pada
tangan dan kaki akibat efek vasokontriksi
– Gangguan sistem syaraf : pusing, vertigo,
gangguan psikis dan halusinasi
Claviceps purpurea
• Alkaloid ergot bekerja pada reseptor a-
adrenergik, dopaminergik dan
serotonergik
• Ergot sejak abad 16 digunakan sebagai
penginduksi kontraksi uterus saat
kelahiran.
• Efek oksitosin (stimulasi otot uterus)
hingga saat ini masih dipakai terutama
ergometrin yang digunakan setelah
operasi Caesar untuk mengurangi
pendarahan.
6. Piridin
Nikotin dan anabasin adalah alkaloid piridin dari
tembakau.
• Pada keduanya selain ada struktur piridin juga ada struktur
pirolidin (pada nikotin) dan piperidin (pada anabasin).
• Tembakau (Nicotiana tabacum, Solanaceae)
mengandung (-) -nikotin 0,6-9%, anabasin dan nornikotin.
• Nikotin ini bersifat seperti minyak, mudah menguap.

Didalam daun, nikotin ini berbentuk garam dengan asam
malat dan sitrat.
• Dosis kecil  nikotin menstimulasi sistem pernafasan
• Dosis tinggi  dapat menyebabkan depresi sistem
pernafasan. Terdapat korelasi yang cukup signifikan
antara kebiasaan merokok dengan kanker.
7. Turunan xantin (basa purin)
• Turunan purin yg terkenal : kafein, teobromin
dan teofilin.
• Kafein banyak dijumpai pada teh, kopi dan
kola, memberikan efek stimulan SSP terbaik
dan sebagai diuretik. Kafein biasa digunakan
bersama obat analgesik.
• Teofilin lebih banyak digunakan sebagai
relaksan otot khususnya untuk asma.
• Teobromin adalah komponen utama dari buah
coklat atau produk dari coklat.
Jumlah kafein pada satu cangkir minuman :
• kopi : 30 – 150 mg (rata-rata 60-80 mg)
• kopi instan : 20 – 100 mg (40-60 mg)
• kopi yg diambil kafeinnya : 2 – 4 mg
• teh : 10 – 100 mg (40 mg)
• coklat : 2 – 50 mg (5 mg)
• kola : 25 – 100 mg
Maksimal kafein dapat dikonsumsi 1 g/hari
Sedangkan dosis letalnya adalah 5-10 g.
►Kopi
• Coffea arabica, C. canephora, C. liberica,
(Rubiaceae). Biji kopi  1-2% kafein dan sedikit
teofilin dan teobromin. Umumnya terikat asam
klorogenat (7%), saat disangrai kafein dibebaskan.
Asam klorogenat  asam kafeat dan asam kuinat.
►Teh
Daun Thea sinensis (Camellia sinensis)
(Theaceae).
• Teh hitam : fermentasi oksidasi enzimatik dari
polifenol
• Teh hijau : segera dikeringkan setelah dipetik
untuk mencegah proses oksidasi/fermentasi.
• Teh Oolong : hasil semifermentasi.
►Kola
• Biji Cola nitida dan C. acuminata (Sterculiaceae)
kafein 3 %, teobromin 0,1%, sebagian terikat tanin. Saat
pengeringan, terjadi oksidasi polifenol  warna merah
dan membebaskan kafein. Digunakan untuk minuman
Coca-cola dan Pepsi-cola.
►Coklat
• Sangat penting secara komersial. Biji coklat
(Theobroma cacao) difermentasi & disangrai untuk
melepaskan teobromin dari kulit biji dan
memperoleh bau khas coklat.
• Biji coklat mengandung 35-50% minyak (cocoabutter
atau minyak teobroma), 1-4% teobromin, 0,2-0,5 %
kafein, tanin serta minyak atsiri. Minyak teobroma atau
pasta coklat diperoleh dengan cara panas berupa
padatan putih dan rasa coklat lemah. Untuk basis
supositoria
8
Terpenoidal alkaloids
Aconite alkaloids Aconitine, molecular formula
C34H47NO11, is an example of an aconite
alkaloid. It is soluble in organic solvents, e.g.
CHCl3 and C6H6, and slightly soluble in
alcohol or ether, but insoluble in water.
Aconitine is an extremely toxic substance
obtained from the plants of the genus
Aconitum (family Ranunculaceae), commonly
known as ‗aconite‘ or ‗monkshood‘. It is a
neurotoxin, and used for creating models of
cardiac arrhythmia.
Steroidal alkaloids
These alkaloids have a core steroidal skeleton as part of the
molecule, e.g. solanine. There are a number of structural varieties
that exist in steroidal alkaloids. Following discussion is just on a few
selected steroidal alkaloids.
Solanine is a poisonous steroidal alkaloid, also known as
glycoalkaloid, found in the nightshades family (Solanaceae). It is
extremely toxic even in small quantities. Solanine has both fungicidal
and pesticidal properties, and it is one of the plant‘s natural defences.
Solanine hydrochloride has been used as a commercial
pesticide. It has sedative and anticonvulsant properties, and has
sometimes been used for the treatment of asthma, as well as for
cough and common cold. However, gastrointestinal and
neurological disorders result from solanine poisoning.
Symptoms include nausea, diarrhoea, vomiting, stomach cramps,
burning of the throat, headaches and dizziness. Other adverse
reactions, in more severe cases, include hallucinations, loss of
sensation, paralysis, fever, jaundice, dilated pupils and
hypothermia. Solanine overdose can be fatal.
Samandarin, molecular formula C19H31NO2, is the major

steroidal alkaloid of the skin glands of the fire salamander
(Salamandra salamandra), and is extremely toxic. The toxicities of
samandarin include muscle convulsions, raised blood pressure
and hyperventilation.
General properties:
1) Most alkaloids are crystalline comp. odorless,
colorless, non volatile, they contain C, H, N, O, few
oxygenated alkaloids  non volatile liquid. e.g:
pilocarpine, pelletierine.
2) Alkaloids lack oxygen in their structure usually
occur as volatile liquid. e.g: coniine, nicotine.
3) Very rare colored alkaloids e.g: berberine,
colchicine (yellow).
4) Being basic, alkaloids  Salts
Alkali hydroxi
Free base
5) Some alkaloids neutral or slight acidic reaction
can not form salts. e.g: ricinine, theophylline.
6) Amphoteric alkaloids contain-COOH (narceine)
or -OH (morphine)
7) Weak basic alkaloids e.g: caffeine, narcotine,
papaverine, piperine from salts  are unstable.
Solubility of alkaloids:
In fact, the differences in solubility between alkaloids and
their salts strongly facilitate the process of isolation and
separation of the alkaloids from plant and from non-
alkaloidal subst.
The general role of solubilily of alkaloids :

- Free bases are soluble in organic solvent, sparingly
soluble in H2O.
- Alkaloids salts are soluble in H2O but sparingly soluble in
organic solvent.
Exception to this generalization:
1) Caffeine (base) extracted from tea by H2O.
2) Colchicine (base) soluble in acid, neutral or
alkaline H2O.
3) Morphine (base) is sparingly soluble in ether.
4) Ephedrine and pilocarpine (base) are soluble in
H2O.
5) Labeline HCl (salt) and apoatropine HCl (salt) are
soluble in CHCl3.
Optical Isomerism:
- Alkaloids are optically active comp.(many of them
contain at least one asymmet C-atom).
-The pharmacological activities are usually more

active laevorotatory isomers than the dextrorotatory
e,g: D(-)- ephedrine 3 fold stronger than D(+)-
ephedrine (3:1) for arterial B.p.
Some exceptions:
d-tubocurarine (dextrorotatory) is the medicinally
active.
2. Penggolongan
Extraction
• Alkaloids can be extracted from their

sources by treatment with acids (usually
hydrochloric acid or sulfuric acid, though
organic acids such as maleic acid and
citric acid are sometimes used).
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:
1- Reagents that form double salts:
a- Mayer‘s Reagent: Potassium Mercuric Iodide.
b- Dragendorff‘s Reagents: Potassium Iodobismethate.
c- Gold Chloride.
2- Reagents Containing Halogens:

a- Wagner‘s Reagent: Iodine/ Potassium Iodide.
3-Organic Acids:
a- Hager‘s Reagent: Picric Acid
b- Tannic Acid.
4- Oxygenated High Molecular Weight Acids:

a- Phosphomolybdic acid
b- Phosphotungestic acid
c- Silicotungestic Acid
• Colour Reagents:
1- Froehd‘s Reagent: Phosphomolybdic acid

2- Marqui‘s Reagent: Formaldehyde/ Conc. H2SO4
3- Mandalin‘s Reagent: Sulphovanidic acid
4- Erdmann‘s 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. H2SO
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.
Plant material and solvent
Organic solvent dissove Alkaloids

Organic solvent dissove Impurities
Extract
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
• The best way for their extraction is steam
distillation.
• Plant material + water + Fixed alkali

Heat
steam contain
alkaloids received in
acidic solution.
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.
Separation of Alkaloidal
Mixtures:
 Fractional Crystallization:
Ephedrine & Pseudoephedrine Oxalates
Crystallization from water
Ephedrine Oxalate Pseudoephedrine Oxalate

Crystals Solution
Atropine & Hyoscyamineine Oxalates
Crystallization from
Acetone/Ether
Atropine Oxalate Hyoscyamine Oxalate

Crystals Solution
 Preparation of Derivatives:
Separation of Primary, Secondary and Tertiary Alkaloids.
O
Mixture + p-toluenesulphonyl chloride Cl S
Add HCl and filter O
Filtrate Precipitate
tertiary alkaloids as salt 1ry alk derivative 2ry alk derivative
(no reaction with reagent
O
O
R-N S
R-HN S keto form R
O
O
insoluble in alkalis
acidic hydrogen
OH
R-N enol form
S
soluble in alkalis
O
NaOH, filter
Filtrate Precipitate
1ry alk derivative 2ry alk derivative
 Fractional Liberation:
Atropine & Hyoscyamine & Hyoscine

the form of HCl salts
1- Alkalinize by NaHCO3 pH 7.5

2- Extract with Ether
Ether Aqueous layer

Hyoscine free base Atropine & Hyoscyamine HCl
(pKa = 6.2) (pKa = 9.3)
• Fractional Distillation:
e.g. Separation of Nicotine and Anabasine
• Chromatographic Separation.
General tests for alkaloids:
1) Alkaloidal precipitating reagents:
A) Mayer‘s reagent. B) Wagner‘s reagent.
C) Hagar‘s reagent D) Gold chloride
E) Dragendorff‘s reagent.
2) Alkaloidal coloring reagents:

(The rough dehydration or oxidation of the alks)
A) Marquis reagent. B) Mandalin R.
C) Froehed‘s reagent D) Erdmann‘s R.
3) Special tests for alkaloids:

Used to identify or to confirm the identity of the alks.
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.
• 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.
• Colourimetric Method:
e.g. Morphine + NaNO2/HCl
Ergot + p-dimethylaminobenzaldehyde
• Spectrophotometric Methods.
• Polarimetric Method.
• Fluorimetric Method.
• Chromatographic Methods
A. Extraction:
In the following a laboratory routine for QA

extraction which has been successfully used
several thousand times .
Stability of alkaloids:
It is important to ensure that, as far as possible,

alkaloids isolated from plants or other sources are
qualitatively and quantitatively the same as those
which were present in the original material.
There are several ways in which alkaloids might be

altered during isolation procedures.
Alkaloids may be changed by heat or by

enzymeic action when harvested plants are stored
and dried.
Bull and call. Reported :
the loss of 50-80% of alkaloids during drying of
some, but not all, Crotalaria and Heliotropium
species [62].
On the other hand, Pderson [63] compared the

alkaloid content of fresh leaves with that of dried
leaves from several species of the Boraginaceae;
allowing for the loss in weight on drying there
were no changes in total alkaloid content,
suggesting that enzymatic oxidation or reduction of
alkaloids could take place during the early stages of
drying.
If fresh green plant material cannot be extracted
straight away, it should be derived as quickly as
possible to minimize enzyme action and fungal
decomposition.
A warm environment, with plenty of circulating

air, is best, but direct sunlight should be avoided
because of possible overheating, the alkaloids are
not especially light-sensitive.
After drying, the plant should be stored in a cool

place [8].
Fresh plant material:
It is common practice to dry plant material prior to
extraction.
Since QA may be modified during the drying

process (e.g. hydrolysis of ester alkaloids), it is
recommended that material is freeze-dried (which
avoids recompart- mentalisation and thus enzymic
breakdown processes) or that fresh material or
material stored frozen at -20°C is used.
The extraction of QA is based on the fact that the
free alkaloidal base does not dissolve in water
but in an apolar organic solvent, such as ether or
methylene chloride.
In contast, the charged alkaloids (most are charged

molecules under acidic conditions) are soluble in
water but insoluble in apolar organic solvents.
Both the free base and the salts dissolve in

MeOH. about 15 ml 0.5M HCI.
For most analytical purposes it is convenient to take 5
or 10g of the fresh or frozen material and to
homogenise it in about 15 ml 0.5M HCI.
Homogenisation can be performed by pestle and

mortar (use quartz sand for grinding), with a Waring
blender or an Ultraturrax.
Great care should be taken that the mortar, blender or

Ultraturrax is cleaned rigorously after use in order to
avoid contamination (QA stick to surfaces!).
The homogenate is transferred to a beaker and is left
standing for at least 30 min (overnight incubations are
also appropriate). In the next step the homogenate is
centrifuged for 10 min at 5000 to 10 000 x g and the
clear supernatant is collected.
For quantitative work the pellet is resuspended in 0.5M

HCI and centrifuged again.
Both supernatants are then pooled, made alkaline by

4M NH4OH (pH 12–14), and left standing for 15 min.
It is recommended that solid phase extraction be used
as the next step, since this procedure can easily be
standardised and applied to the extraction of many
samples concomitantly.
Chemelut (Analytichem) or Extrelut (Merck) are

modified kieselgur powders which have a high
adsorption capacity for water (c. 14g bind 20ml water).
Commercial extraction columns (Merck or Analytichem)

(which can be reused after thorough cleaning) are filled
with c. 14g matrix material.
Up to 20 ml of the homogenate (determine the exact
volume!) can be applied on the column. It will take c. 10
min for the homogenate to be adsorbed the collumn
matrix.
The column is then eluted 2 to 3 times with 20ml

methylene chloride.
The eluate is collected in a flask and evaporated in a

rotavapor under reduced pressure at 40°C.
When the solvent has been evaporated the crude
alkaloid extract is dissolved in a small volume of
methylene chloride and transferred quantitavely to a
sample vial.
The solvent can be evaporated in these vials either by

delivering a steady stream of nitrogen gas or just by
leaving the samples overnight in a fume cupboard.
Then the vials are closed and stored if possible at 4°C

or even better at –20°C until analysis.
Alternatively, extraction can be performed by liquid-
liquid extraction: methylene chloride is added to the
alkaline homogenate and mixed in a shaking funnel.
The lower organic phase is collected.
The process is repeated at least twice.
The organic phases are pooled and processed as

described above.
Depending on the material, emulsion will be

encountered using the liquidliquid precedure but not
with the solid- phase extraction [27].
Seeds and dried material:
Dry material, which needs to be milled into a fine
powder prior to extraction, can be extracted by two
procedures.
a) About 500mg of plant material is suspended in 15

ml 0.5 M HCl and left standing under continuous
stirring (magnetic stirrer) for at least 3h. The
following steps are identical to those outlined above.
b) About 500mg or more of plant powder are extracted

with MeOH for several hours, which is conveniently
performed with a Soxhlet apparatus.
The methanolic extract is then evaporated to dryness
under reduced pressure in a rotavapor. For some
applications (HPLC, TLC) this extract can be used
directly.
Normally, these extracts need to be further purified:

the dried residue is dissolved in 15ml 0.5 M HCl and
extracted at least three times with methylene chloride
(until a clear organic phase is obtained) in a shaking
funnel to remove lipid material.
Then the aqueous phase is made alkaline with 4N

NH4OH and extracted with methylene chloride using
either solid-phase extraction or liquid-liquid extraction
as described above [27].
B. Detection of Alkaloids:
Numerous reagents are recommended for simple '
spot test ' identification of alkaloids, most working
best in a dilute aqueous acid solution of the
alkaloid.
Pride of place among these must go to

Dragendorff‘s Reagent but there are many others,
such as iodoplatinic acid, Mayer‘s Reagent,
Hagar‘s Reagent and Wagner‘s Reagent.
Mixing a drop of extract with a drop of the Reagent

will lead to the development of a precipitate if
alkaloids are present.
Great care must be taken with these spot tests
because, while alkaloids will generally be
characterised by strong reactions, other types of
natural products are able to react to some degree
(flavonoids, coumarines, tannins).
So-called ' false positives ' can be eliminated by

taking the alkaloid-containing acid extract, making it
basic and extracting the alkaloid into an organic
solvent.
DETEKSI
CHCl3/NH3
• saring
• Uji Dragendorf
• Uji Meyer
• Uji Wagner
DETEKSI
Pereaksi tetes:
• Pereaksi Dragendorf: larutan senyawa
kompleks Bi(NO3)3/KI dalam asam
Positif  endapan jingga kemerahan
• Pereaksi Meyer: larutan senyawa
kompleks HgCl2/KI
Positif  endapan putih
• Pereaksi Wagner: larutan senyawa
kompleks KI/I2
Positif  endapan coklat
endapan jingga kemerahan
endapan putih
This is followed by separation of the organic phase
and its back extraction into acid again.
If the new acid extract still gives a positive reaction

then the presence of alkaloid base is confirmed;
although if the alkaloid had been non-basic or
quaternary and hydrophilic it would be lost.
In addition to these general colour reagents there are

many others that are said to be specific to certain
alkaloid types.
Today, their use is very limited [2].

A. Extraction:
In the following a laboratory routine for QA

extraction which has been successfully used
several thousand times .
QUALITATIVE AND
GENERAL PROPERTIES
WHAT IS THE ALKALOIDS'S GENERAL PROPERTIES?
• Alkaloids are generally bitter in taste and are optically active.

• Most alkaloids are crystalline,colourless,non-volatile solids,insoluble
in water , but soluble in polar organic solvents.
• Some alkaloids are water-soluble liquids
(for exemple coniine,hygrine,nicotine).
• Few alkaloids are coloured (for exemple berberine is yellow).
• Nearly all basic alkaloids form salts with inorganic acids,which are
soluble in water and insoluble in organic solvents.
The knowledge of solubility of alkaloids and their salts is utilized in

their isolation.
HOW ARE ALKALOIDS DETECTED?
• Alkaloids yield precipitates or characteristic colours when

reacted with solutions of molybdic acid,picric
acid,chloroplatinic acid,potassium mercuric iodide
etc.Therefore the presence of an alkaloids in a plant is
detected by using various reagents such as Mayer's
reagent,Wagner's reagent,Dragendorff's reagent,Hager's
reagent,chloroplatinic acid, Frohde's reagent etc.
HOW ARE ALKALOIDS DETECTED?
• These precipitates may be amorphous or crystalline and are

of various colours:
cream(Mayer's)
yellow(Hager's)
reddish-pinkish-brown(Wagner‗s and Dragendorff‘s )
※Caffeine and some other alkaloids do not give these

precipitates.
• Pemisahan alkaloid dengan memanfaatkan
sifat kelarutannya yang berbeda sebagai
bentuk garam dan bentuk bebas/basanya:
– Dengan asam membentuk garam (mis. asam

sulfat, klorida, fosfat dll), larut dalam air,
tidak larut dalam pelarut organik.
– Dalam bentuk basa, kurang larut dalam air,

larut dalam pelarut organik.
Extraction
• Alkaloids can be extracted from their

sources by treatment with acids (usually
hydrochloric acid or sulfuric acid, though
organic acids such as maleic acid and
citric acid are sometimes used).
DETEKSI
CHCl3/NH3
• saring
• Uji Dragendorf
• Uji Meyer
• Uji Wagner
ISOLATION OF CAFFEINE
FROM TEA LEAVES
Procedure:
1. Carefully remove the tea from the first teabag (regular),
weigh it and record the mass of the tea. Put the tea back
into the back and re-staple it.
2. Place the teabag and about 40 mL of distilled water

into a 250 mL beaker. Cover it with a watchglass and heat
to a gentle boil. Boil for 5 minutes.
3. Decant the liquid into a 150 mL beaker. Add about 20

mL more water to the teabag and boil again for 5
minutes.
4. Decant the liquid into the the 150 mL beaker (with the
first liquid). Use a glass rod to gently squeeze out as
much liquid as possible (without breaking the bag). Add
about 1 g of Na2CO3, swirl to dissolve, and allow the
liquid to cool to room temperature.
5. Transfer the liquid to a 125 mL separatory funnel

(supported with a ring clamp). Add 10 mL of methylene
chloride, shake gently (vent frequently) and allow mixture
to separate. Collect the organic layer in a 100 mL beaker
(label and weigh the beaker first and record the mass).
Add another 10 mL of methylene chloride to the
separatory funnel, shake, separate and collect in the 100
mL beaker. *If an emulsion forms, try adding a little NaCl
(1 or 2 scoops) and shake gently.
6. Discard the aqueous layer and rinse out the separatory
funnel. Allow the methylene chloride to evaporate while
you extract your second teabag.
7. Repeat the above procedures with the second teabag

(decaffeinated), collecting the methylene chloride (step 5)
in a 50 mL beaker.
8. If the methylene chloride from the first extraction

(regular tea) is all evaporated, then weigh the beaker +
caffeine and record the mass. If it‘s not dry, run a stream
of air over it to complete drying before weighing
(BE VERY CAREFUL THE AIR PRESSURE DOESN‘T
CAUSE SPLASHING).
9. After you record the mass, scrape out a small amount

of caffeine and place it in a small test tube. Add about 0.5
mL of methylene chloride.
10. Prepare a TLC plate by drawing a line about 1 cm
from the bottom of the plate, and then marking 3 places
on the line for spotting. Obtain a TLC chamber and two
capillary tubes. Place about 8 mL of TLC solvent (1%
glacial acetic acid in acetone) into the chamber.
11. Spot the plate with your samples. First, spot the left
and center marks with your first tea extract (one spot
each). Next, with a new capillary tube, spot the second
tea extract (from the 50 mL beaker) on the right-hand
mark. Spot 3 or 4 times, allowing the spot to dry between
each (this solution is more dilute). Then spot the second
tea extract in the same way over the center mark, right on
top of the first spot. Label the spots under the marks.
12. Develop the TLC plate and mark the solvent line
before drying it. Observe the plate under short-wave UV
light and circle any spots you see. Note the appearance
of the spots (bright, dark, colored, etc.). Make a sketch of
the plate (with observations) in your notebook. Measure
the Rf of each spot.
13. If the methylene chloride from the second extraction

(decaffeinated tea) is all evaporated, then weigh the
beaker + caffeine and record the mass. If it‘s not dry, run
a stream of air over it to complete drying before
weighing.
14. Calculate the mass of the caffeine obtained from each

tea, and then calculate the % by mass of caffeine for each
tea.
Waste Disposal:
Teabags can go into the trash.
Aqueous solutions should go into the aqueous waste

container (they contain base).
Any organic chemicals should go into the organic waste

container.
Capillary tubes go into the glass waste (not the trash).

Report:
1. In your notebook, make tables for your results. One
should include the brand of tea, regular or decaffeinated,
mass of tea, mass of caffeine and % by mass of caffeine
in the tea for each tea tested (two in this case). The other
should include the Rf values and appearance for each
spot for each of the three lanes on your TLC.
2. Write a brief discussion of the experiment. In this

section you should summarize your results (brand,
regular or decaffeinated and % by mass caffeine) and
note any interesting observations (odor, color etc.). Also
summarize your TLC results and discuss their relevance.
Make any possible conclusions about the experiment
(successful vs. unsuccessful and reasons why; compare
your results to expected results).
3. Answer the post-lab question.
4. Make a copy of your lab notebook pages (do not tear

out the originals) and attach the question sheet with your
answers.
5. Staple the discussion, post-lab question notebook

page copies and upcoming pre-lab together.
Post-Lab Questions:
1. Before extracting the caffeine with methylene chloride,
you added Na2CO3 to the aqueous portion. Explain why.
Also, write the chemical reaction that would occur
between phenol and NaOH and incorporate it into your
explanation.
2. Caffeine is soluble in methanol, as well as methylene

chloride. Why couldn‘t you use methanol to extract the
caffeine from the aqueous portion?
Caffeine Content of Some Foods and Drugs
Coffee 80-125 mg per cup

Decaffeinated Coffee 2-4 mg per cup
Tea 30-75 mg per cup
Cocoa 5-40 mg per cup
Milk Chocolate 6 mg per oz
Baking Chocolate 35 mg per oz
Coca-Cola 46 mg per 12 oz can
Anacin, Midol 32 mg per tablet
Excedrin, extra strength 65 mg per tablet
Dexatrim, Vivarin 200 mg per tablet
Dristan 16 mg per tablet
No-Doz 100 mg per tablet
FROM TEA LEAVES
FROM TEA LEAVES
ISOLATION AND
PURIFICATION
The differences in solubility between alkaloids and their salts
provide methods for the isolation of alkaloids from the plant and
their separation from the non-alkaloid substances.
• For many alkaloids are basic and occur as salts of 2-hydroxybutane-

1,4--dioic acid (malic acid),or of 1,3,4,5-tetrahydroxy cyclo hexane
(quinic acid).They can thus be extracted into acid solution using
aqueous HCl, tartaric, or citric acids. Essentially neutral alkaloids like
colchicine or piperine remain in the organic phase ,while most other
alkaloids are isolated after basification and extraction into ethyl
acetate.
• The almost invariably subsequent purification of the crude alkaloid

mixtures is effected by chromatography using silica or
alumina,and then recrystallization of the partially purified
compounds from solvent systems like aqueous ethanol
,methanol/chloroform,or methanol/acetone.
• Pemisahan sesama alkaloid Rauwolfia
berdasarkan kebasaannya :
– serpentin paling kuat sifat basanya,

– ajmalin dan senyawa sejenisnya
kebasaannya menengah
– reserpin, resinamin, deserpidin dan
ajmalisin merupakan basa lemah
(G.A. CORDELL)
Bahan tumbuhan
eter minyak bumi

ii) Pemekatan
tartrat 2%

Partisi dgn EtOAc

Diekstraksi
dgn CHCl3

ISOLASI ALKALOID
ISOLASI: skema umum
Pelarutan dengan
cara perendaman
EKSTRAKSI
Partisi asam-basa
Pengambilan
semua FRAKSI ALKALOID
komponen
yang terlarut
• Partisi
Pengambilan • Kromatografi
hanya FRAKSINASI
komponen
alkaloid
• Kromatografi
Penyederhana • Kristalisasi
an jumlah PEMURNIAN
komponen
Mendapatkan masing-
masing komponen murni
PROCEDURE USED FOR ALKALOIDS ISOLATION
• Alkaloids with low molecular weight or volatile liquid

For example:nicotine ,sparteine and coniine Distillation
An aqueous extract of plant is made alkaline with

caustic soda or sodium carbonate and alkaloid is
distilled off with steam.The distillate is extracted with
solvent to isolate the desired alkaloid.
(G.A. CORDELL)
Bahan tumbuhan
eter minyak bumi

ii) Pemekatan
tartrat 2%

Partisi dgn EtOAc

Diekstraksi
dgn CHCl3

• Alkaloids with few exceptions salts of alkaloids are water-soluble
Powdered plant material
First extracted with petroleum ether
(for the removal of fats)
Then extracted with methanol or
ethanol
Dark solution
Using Evaporated to dryness
column chromatography
Residue
or counter-current
Add water, then the mixture is acidified
distribution method.
to pH 2 and extracted with ethyl acetate
or ether ( for the removal of non-basic
molecules )
Fractional Aqueous solution (see next page)
crystallization Purified by
of salts Basified to pH 10 and extracted
with ethyl acetate or ether
Water insoluble organic layer ( Evaporated to dryness)
(crude alkaloid)
• Alkaloids with few exceptions salts of alkaloids are water-soluble
Aqueous solution
Precipitated with ammonium Peinecke's
(H[Cr(NH3)2(SCN)4]) solution
The precipitated salt
Ion exchange chromatography
Eluent
freeze-drying
Water-soluble alkaloids
Isolation and purification of an alkaloid from a plant material may not always be a
simple procedure as described above because some alkaloid bearing plants contain
a complex mixture of several alkaloids.Therefore,the isolation of a pure alkaloid from
these plants may sometimes due to the repeated use of chromatography or
crystallizations.
ISOLASI: alkaloid dari Eurycoma longifolia Jack
Serbuk akar (10,5 kg)

• ekstraksi dengan MeOH
Residu Ekstrak MeOH

• partisi ke dalam CHCl3 – n-butanol
Fraksi CHCl3 Fraksi n-butanol
• fraksinasi dengan KVC

eluen: heksana-EtOAc, EtOAc, EtOAc-MeOH
F - 1 F - 2 F - 3 .... F - 12
Pemurnian masing-masing fraksi dengan teknik kromatografi

menghasilkan 22 senyawa alkaloid
ISOLASI: alkaloid dari Litsea tomentosa Blume
Tahap pertama: pembuatan fraksi alkaloid total
Serbuk kering kayu batang (2 kg)
• ekstraksi dengan MeOH
Residu Ekstrak MeOH

• penguapan pada tekanan rendah
Ekstrak MeOH kental
• partisi ke dalam larutan 5% as. sitrat dan EtOA
Lapisan air Fraksi EtOAc (netral)
• + NH3 sampai pH ~9-10

• ekstraksi dengan EtOAC
Lapisan air Fraksi EtOAc

ANALISIS KLT
(alkaloid total, 7.4 g)
ISOLASI: alkaloid dari Litsea tomentosa Blume
Tahap kedua: fraksinasi dan pemurnian alkaloid
Fraksi alkaloid (7,4 g)
• fraksinasi dengan KVC
eluen: heksana-EtOAc, EtOAc, EtOAc-MeOH
Kromatografi
vakum cair (KVC)
F - A F - B F - C F - D F - E
• fraksinasi dengan
KVC
• pemurnian dengan
kromatografi radial
Kromatografi radial
12/16/2016 4:48:14 PM Phytochemistry 242
Chemical content of Curcuma domestica (kunyit)
and C. xanthorriza (temulawak)
O O
C.domestica
H3CO OCH3
C.xanthorrhiza HO OH
Curcumin
O O
H3CO
Xanthorrhizol
HO OH
Demetoxycurcumin
O O
HO OH
Bisdemetoxycurcumin
O OH
Fluorene Fluorenone Fluorenol
a) Which one of these compounds is the least polar?
b) Which one of these compounds is the most polar?
c) What would be the relative order of separation on

the TLC plate remembering that CH2Cl2 is not very
polar?
Alumina and cellulose are the two most often used
nonsilica-based sorbents.
Alumina has been used to separate compounds such as

fat-soluble vitamins, alkaloids, and antibiotics.
Cellulose-based ion-exchange layers have wide

application for sulfonamides, nucleic acids, and steroid
sulfates
Alumina
During the last 20 years, the chromatographic use of
aluminum oxide has steadily decreased even though
alumina partially or totally overcomes the difficulty arising
from the relatively low pH stability of silica.
Alumina oxide is stable over a broader pH range (2-12)

than silica gel but generally has a lower specific surface
area, typically 70 m2/g. Like silica, alumina can also act like
an ion exchanger. Alumina has amphoteric properties
with both cation and anion-exchange properties ranging
over a broad pH range. At solvent pH values below that of
the pK of the alumina surface, the alumina surface has a
positive net charge. At a higher pH, the surface charge is
negative (Figure 5.1). The zero-point charge (no net charge)
of alumina occurs at a pH of 9.2 but can be shifted to 6.5 in
acetate buffer and 3.5 in citrate buffer.
Similar to what has occurred with silica, alumina-based
reversed phase has been prepared by the free-radical
polymerization of cross-link polymers to alumina particles or
by chemically bonding alkyl groups to alumina.
Cyano bonded to alumina has been shown to be useful in

resolving various pencillins, cephalosporins, macrolide
antibiotics, and tricyclic antidepressants.
Neutral
alumina
DETEKSI
Pereaksi semprot pada pelat KLT:
• Pereaksi Dragendorf: larutan senyawa kompleks Bi(NO3)3/KI dalam asam
Positif  noda berwarna jingga kemerahan
Some common spray reagents for detecting
alkaloids.
No Reagent Composition
1 Dragendorff (a) 0.85g bismuth (III) nitrate in 10ml
Reagent glacial acetic acid and 40ml water.
(b) 8g potassium iodide in 20ml water.
Mix (a) and (b) in equal portions to use
as spray or spot test reagent.
2 Cerium 0.1g cerium (IV) sulphate in 4ml water.
sulphate Add 1g trichloroacetic acid, boil and
add dropwise 97% sulphuric acid until
solution clarifies.
Heat to produce colours.
Some common spray reagents for detecting
alkaloids.
No Reagent Composition
3 Cobalt (II) Ammonium thiocyanate (3g) and
thiocyanate cobalt (II) chloride (1g) in 20ml water.
4 Iodoplatinic (a) 5% hexachloroplatinic (IV) acid

acid solution
(b) 10% potassium iodide solution.
Mix 5ml (a) with 45ml (b).
5 Sodium 1% sodium tetraphenyloboron
tetraphenyl solution in ethyl methyl ketone
saturated with water.
boron
C. Chromatography:
1-Thin Layer Chromatography:
Since most plants contain complex mixtures of up to
30 or more individual alkaloids, Thin Layer
Chromatography (TLC), with its limited resolution
power, is helpful primarily for initial analysis of the
major alkaloids and for preparative work. A
quantitative TLC system has been described by
Karlsson and Peter [64].
The following solvent systems are adequate for silica

gel G plates:
a) MeOH –28% NH4OH (65:1).
b) CHCL3–MeOH–28% NH4OH (85:15:1).
c) Cyclohexane –diethylamine (70:30).
In Table I-2 Rf Values of some representative QA are
tabulated.
They can be visualised with ' Dragendorff‘s ' reagent

as orange red spots; KI/I2 produces dark brown
stains.
Plates need to be dried and heated (20min at 110°C)

prior to visualisation in order to remove the ammonia
or diethylamine which interfere with the colour
reaction [27].
System 1 : MeOH –28% NH4OH (65:1) .

System 2: CHCL3–MeOH–28% NH4OH (85:15:1).
System 3 : Cyclohexane –diethylamine (70:30).
Table I-3: Selection of TLC Rf values [27].
No Alkaloid System 1 System 2 System 3
1 Lupinine 0.25 0.36 0.52
2 Camoensidine 0.16 0.31 0.39
3 Lupanine 0.30 0.69 0.51
4 Sparteine 0.07 0.93
5 Multifloine 0.27 0.35 0.26
6 Aphylline 0.60 0.80 0.52
7 Angustifoline 0.41 0.63
8 13 -a- 0.51 0.69 0.50
Isolupanine
9 Anagyrine 0.49 0.66 0.30
10 Matrine 0.26 0.52 0.35
11 Ammodendrine 0.17 0.23 0.37
2-Liquid Chromatography:
(a) Low-pressure Chromatography.
For preparative work QA can be separated by

liquid chromatography on silica (70-230 or 230-400
mesh, Merck) with the following solvent systems:
CH2CL2–MeOH –25% NH4OH (70:30:1) [65] or
CH2CL2–MeOH –25% NH4OH (90:9:1) [66] or
diethyl ether–MeOH–25% NH4OH (100:30:1) [66].
Arslanian and coll. [67] used a basic AL2O3 column

(Merck, Type T) with solvents of increasing polarity,
starting with ether, then 10% MeOH in ether, 50%
MeOH in ether, and finally MeOH.
(b) High Performance Liquid Chromatography.
High performance liquid chromatography (HPLC) has been

employed to the analysis of QA mixtures.
Saito and coll. [68] Used an Inertsil ODS (5μm( column (150
x 4.6mm( with the solvent system acetonitrile– mM
potassium phosphate buffer (pH 5.5) (1:9).
Murakoshi and coll. [66] used LiChrosorb Si 100 (Merck;
10μm, 0.3 x 50cm) with the solvent system 15% MeOH in
Et2O–H2O–28% NH4OH (500:10:3).
Saito and coll. [68] separated 22 of these alkaloids on

LiChrosorb Si 60 (5μm) (250 x 4.6mm) with the solvent
systems 15% or 25 % MeOH in diethyl ether–5% NH4OH
(25:1), or 50% MeOH in diethyl ether–5% NH4OH (25:2).
Detection can be performed using refractive index
(RI) or UV detectors.
Since most QA have no sensitive chromopore, UV

detection is performed between 205 and 220nm.
α-Pyridone alkaloids (Fig. I-1, type 4), such as

cytisine or anagyrine, have an aromatic ring and
can be monitored at 310nm [68].
Using preparative columns, HPLC is a convenient
technique for isolating sufficient amounts of individual
alkaloids (only spateine, lupinine, α-isolupanine and
cytisine are commercially available) for spectroscopic
or biochemical purposes [69].
Wink have successfully employed a semi-preparative
column (300 x 7.8mm; 10μm material, μ-Bondapak
C18; waters), using an isocratic system with the
solvent system 0.01M phosphate buffer – 20%
MeOH – 20% isopropanol.
Murakoshi and coll. [66] and Ohmiya and coll. [69]
used Lichrosorb Si 100 (Merck ; 10μm , 0.3 x 50 cm)
with the solvent system 15% MeOH in Et2O–H2O –
28% NH4OH (500:10:3).
3-Gas –Liquid Chromatography.
Until 1980 most gas–liquid chromatography (GC)
applications to separate QA mixtures involved packed
columns. However , best results can be obtained using
the new generation of fused silica capillary columns
with bonded phases :
The capillary columns (dimensions 15m x 0.23mm to
30m x 0.32mm) have a higher number of theoretical
plates (> 70 000) , which allow the separation of
complex mixtures (Fig. I-2) and even of enantiomers ,
epimeric at C-11 or C-6 , such as sparteine and a-
isospartiene , lupanine and -isolupanine ,13-
hydroxylypanine and 13-epihydroxy-lupanine ,
anagyrine and thermopsine ,13a-tigloyloxylupanine
and 13 -tigloyloxylupanine , and of cis and trans
isomers , such as 13a-Angeloyloxylupanine and 13a-
Tigloyloxylupanine , as well as the trans- and cis-
cinnamic acid esters [70
Furthermore, even nanogram amounts of QA can be
detected by FID (flame ionisation detector) or more
sensitively and specifically by a nitrogen specific
detector (abbreviated PND or NPD).
Therefore, for most biochemical and phytochemical

applications, capillary GLC seems to be the method of
choice for QA analysis.
A further convenience is the possibility of performing

GC-MS analyse [27] (section I-2-3-B).
As a liquid phase several silicone derivatives (0.1μm or
1μm films) are employed , good resolutions have been
obtained using DB-1 or DB-5 columns (J & W
Scientific) , but
equivalent products of other manufacturers also work .
Split injection techniques are usually appropriate.
On-column injection does not provide significant

advantages for most applications.
Helium is routinely used as carrier gas, but hydrogen

or nitrogen will also work.
The injector temperature is usually set to 250°C, that of

the detectors to 300°C [71].
Hydroxylated QA, sush as 13-hydroxylupanine or 4-
hydroxylypanine, may be derivatised by TMS prior to
injection to avoid tailing and to achieve better
quantification.
Care should be taken not to use the PND for these

derivatives, since the PND would soon be destroyed
[27].
Some authors give relative retention indices for QA

(e.g. [72]). However, Kovats retention indices (RI) [73]
give better comparative information and are helpful in
identifying
individual alkaliods in a GC profile. Some
representative RI indices are given in Table I-4.
ANALISIS ALKALOID
Analisis alkaloid dari sampel tumbuhan umumnya menggunakan
metoda HPLC atau GC yang digabung dengan MS:
• HPLC-ESI-MS
• HPLC-APCI-MS
• GC-MS
EKSTRAKSI
FRAKSI ALKALOID
ANALISIS
METHODS OF
STRUCTURE DETERMINATION
2. Penggolongan
GENERAL METHODS OF
STRUCTURE DETERMINATION
• In structure determination of alkaloids,a variety of general chemical
methods and more recently physical methods are employed.
• In general,elemental composition is obtained from combustion
analysis and after determination of molecular weight,molecular
formula is calculated.The measurement of optical rotation indicates
the presence of optical activity.
METHODS:
A. Chemical Methods
B. Degradation of Alkaloids
C. Physico chemical Methods
 A. CHEMICAL METHODS
• The alkaloids mostly contains one or more oxygen atoms,which may
be present as hydroxyl,methoxy,methylenedioxy,carbonyl ,carbonyl
ester,lactone,amide,lactam,epoxide groups or ether linkage.
• i) Hydroxyl group
Molecule contains hydroxyl group or -NH group then the number of
these groups can be estimated by acetylation or Zerewitinoff's method.
• Acetylation's method
R-OH + CH3-CO-Cl → R-OCO-CH3
R-NH-R1 + CH3-CO-Cl → R-N(COCH3)-R1
• Zerewitinoff's method
R-OH + MeMgI → R-OMgI + CH4
R-NH-R' + MeMgI → R-N(MgI)-R' + CH4
• If hydroxyl group is present it may be Alcoholic or Phenolic.
Phenolic compounds are soluble in sodium hydroxide and are reprecipitated by carbon
dioxide and give colouration with ferric chloride while alcoholic does not respond to
these tests .
 CHEMICAL METHODS
• ii) Carbonyl group
Ascertained by usual reactions with hydroxylamine, semicarbazide or 2,4-
dinitrophenyldrazine.The carbonyl group may be present as an aldehyde or
a ketone.This distinction can be made from Tollen's reagent and silver
mirror .
• iii) Carboxyl group
Dissolved in bicarbonate or ammonia and reprecipitation with carbon dioxide
indicates the presence of carboxyl group.
The formation of ester on treatment with alcohol in the presence of
dehydrating agent.
• iv) Methoxy group
Use Zeisel's method,which is similar to the Herzig-Meyer method
126 ℃
R-OMe + HI R-OH +MeI AgI
AgNO3
CHEMICAL METHODS
• v) Methylenedioxy group (-O-CH2-O-)
On heating with hydrochloric or sulfuric acid yields formaldehyde.
• vi) Amide, lactam, ester,lactone groups
Be detected and estimated through acid or alkaline hydrolysis.
• vii) Epoxide and ether linkage
Be cleaved by the addition of hydrogen bromide or hydroiodic acid.
• viii) Tertiary methyl group
Estimated by Kuhn-Roth oxidation (K2Cr2O7/H2SO4) to acetic
acid,which is distilled off and titrated against standard base.
-C-Me + K2Cr2O7/H2SO4 → CH3COOH
• ix) Nature of nitrogen
The acetylation or benzoylation can distinguish tertiary amine from
secondary amine,the former being inert whereas the latter gives
acetate or benzoate derivative.
This distinction can also be done by treatment with HNO2 or methyl
iodide or oxidation with 30% hydrogen peroxide.
CHEMICAL METHODS
• ix) Nature of nitrogen
The presence of N-methyl group is often detected by distillation of amine
with sodalime or estimated by the treatment with hydroiodic acid at 150-
300℃ and conversion of methyl iodide produced to silver iodide as
mentioned for estimation of methoxy groups.
• Secondary amine
• >N-H + HNO2 → >N-NO+H2O
• >NH + CH3I → >N-Me + HI
• Tertiary amine
• >N + CH3I → >N+ -Me I-
• >N + H2O2 → >N+-O-+ H2O
• N-Methyl group
• >N -Me + CaO → CH3NH2
AgNO3
• >N -Me + HI → N-H + MeI AgI
 B. DEGRADATION OF ALKALOIDS
The following methods are used to find out structural fragments of

alkaloid molecules:
i. Hofmann exhaustive methylation.

ii. Emde's degradation.
iii. von Braun's method.
iv. Hydrolysis.
v. Alkali fusion.
vi. Dehydrogenation.
 DEGRADATION OF ALKALOIDS
Now we discuss the mainly used methods of Hofmann degradation
and Emde's degradation.
• i. Hofmann exhaustive methylation
• The alkaloids is converted into the quaternary ammonium salt by the
treatment with excess of alkyl halide and the resulting salt is then
converted into the more basic hydroxide by reacting with silver
oxide.subsequently the quaternary hydroxide salt is heated at 200℃
to give an olefin with the elimination of a tertiary amine and water
molecule.
R-CH2-CH2-N+R3 + OH- → R-CH=CH2 + H2O +NR3
• The elimination generally proceeds by an E2 elimination in which the
β-hydrogen and quaternary nitrogen groups are present in the trans-
antiparallel configuration.
• Hofmann's degradation fails in the following cases:
i. with tetrahydroquinoline
ii. with aromatic heterocyclic rings
iii. With compounds having no β-hydrogen atom
 DEGRADATION OF ALKALOIDS
Hofmann degradation fails in the case of derivatives of 1,2,3,4,-
tetrahydrodimethyl quinolium hydroxide.However,Emde degradation
results in cleavage of ring.
• ii. Emde's degradation

• The alkaloid is converted to quaternary ammonium salt by refluxing
with alkyl halide and the resulting salt is subjected to reductive
cleavage by the treatment with sodium amalgam in alkanol or sodium
in liquid ammonia or by catalytic hydrogenation.
• Tetrahydroquinoline and tetrahydroisoquinoline quarternary salts

undergo Emde's degradation.
C. PHYSICOCHEMICAL METHODS
Recently physical methods are used,in conjunction with chemical
reactione to elucidate structure of alkaloids and it is possible to determine
a structure in a matter of days given a few milligrams(or less) of a pure
compound.
• Ultraviolet spectra：Used to indicate the nature of unsaturation or
aromatic rings
• Infrared spectrum：Gives information about many functional groups
• NMR spectroscopy：More versatile for detecting many function groups,
the nature of protons,carbons, heterocyclic rings etc
• Mass spectra：The fragmentation gives the information about molecular
weight and degradation of the skeleton.
• Single crystal X-ray analysis :Offers means for determining or confirming
stereochemistry as well as distinguishing between alternate structures
that appear to fit well for a particular alkaloid.
• optical rotatory dispersion or circular dichroism: Further support for the
stereochemistry
Spektrofotometri UV/VIS
I. Panjang gelombang maksimum (lmax)
• Harga lmax tergantung struktur dan jenis pelarut
II. Absorbansi (A) dan Absorptivitas molar (E)

• Harga A dan E sangat khas untuk masing-masing senyawa
III. Korelasi spektrum dengan sistem elektron

• Penambahan pereaksi geser dapat menunjukkan sistem
elektron dalam struktur tsb
• Kromofor : minimal 3 ikatan rangkap terkonjugasi

• Auksokrom
• Penggeseran panjang gelombang
UV Spectroscopy
II. Instrumentation and Spectra

B. Instrumentation – Sample Handling
7. Additionally solvents must preserve the fine structure (where it is actually
observed in UV!) where possible
8. H-bonding further complicates the effect of vibrational and rotational

energy levels on electronic transitions, dipole-dipole interacts less so
9. The more non-polar the solvent, the better (this is not always possible)
277
UV Spectroscopy
II. Instrumentation and Spectra

C. The Spectrum
1. The x-axis of the spectrum is in wavelength; 200-350 nm for UV, 200-700
for UV-VIS determinations
2. Due to the lack of any fine structure, spectra are rarely shown in their
raw form, rather, the peak maxima are simply reported as a numerical list
of “lamba max” values or lmax
NH2
lmax = 206 nm
O O 252
317
376
278
I-2-3-Characterisation:
A. Ultraviolet and Infrared Spectroscopy:

Only the a-pyridone alkaloids with an aromatic ring A and
the cinnamoyl derivatives have a reasonable chromophore;
thus UV spectroscopy does not normally provide much
information for the identification of QA [74].
Until c. 1970 infrared spectroscopy was an important method

for the identification of QA; such data are found in many papers
[75].
Since MS and NMR are more powerful methods, infrared

spectroscopy has been replaced by these to a large degree.
Optical rotation measurements and circular dichroism have
been employed to determine the absolute configuration of QA
by, among others, [76] and [77].
PENENTUAN STRUKTUR ALKALOID: spektroskopi
Identifikasi unit-unit struktur:
Spektrum UV, IR, NMR (1D dan 2D), dan massa
Contoh:
Spektrum UV dapat digunakan dalam membedakan jenis-jenis kerangka
alkaloid turunan benzilisokuinolin
252 230
CH3O
270 NH
CH3O
279
328 HO
A
315 283
CH3O
N
CH3O
HO
Krikonisin N-Norarmepavin
Panjang gelombang (nm)

Nicotine
Nicotine
Caffeine
BY PLANT OF ORIGIN
HO N
MeO quinine
N CH3
MeO O OH
morphine
OH
HO
MeO belladine
O
MeO N
O
N lycorine
MeO
H
The other three
H
belladine.
H
N CH3 O N CH3
OH
O
MeO
O
tazettine
O daffodils
narcissus
etc
HETEROCYCLIC RING SYSTEMS
LEARN THESE RINGS (plus the ones on the next page)
N N N N
H H H
N N N
N
H H
HETEROCYCLIC RING SYSTEMS (cont)
LEARN THESE ALSO
H
N
N N
quinolizidine pyrrolizidine tropane
N N
N
C C N
N N
H
benzylisoquinoline purine -phenylethylamine

Infrared Spectroscopy
Background
• Absorption of IR light causes changes in the vibrational
motions of a molecule.
• The different vibrational modes available to a molecule
include stretching and bending modes.
• The vibrational modes of a molecule are quantized, so they

occur only at specific frequencies which correspond to the
frequency of IR light.
Background
• When the frequency of IR light matches the frequency of a
particular vibrational mode, the IR light is absorbed, causing
the amplitude of the particular bond stretch or bond bend to
increase.
IR Chart
Characteristics of an IR Spectrum—1-Propanol
IR Absorptions in Hydrocarbons
Hexane has only C-C single bonds and sp3 hybridized C atoms. Therefore
it has only one major absorption at 3000-2850 cm-1.
IR Absorptions in Nitrogen Containing Compounds
The CN of the nitrile absorbs in the triple bond region at ~2250
cm-1.
Nicotine
NH
C-H
Nicotine
NH
Caffeine C= O
Spektrometri Massa
• I. Bobot molekul
• BM genap (jumlah N genap) atau BM ganjil (jumlah N ganjil)
• II. Susunan unsur

• Atom Cl dan Br
• III. Rumus molekul

• Pada spektrometri massa resolusi tinggi puncak-
puncaknya disertai rumus molekul
• IV. Gugus fungsi
• Beberapa gugus fungsi mudah dikenali fragmentasinya
• V. Kerangka molekul
• Beberapa golongan senyawa dapat dikenali
kerangkanya dari fragmentasinya
Interpreting MS
• The only elements to give significant M + 2
peaks are Cl and Br. If no large M + 2 peak
is present, these elements are absent
• Is the mass of the molecular ion odd or even?
• Nitrogen Rule: if a compound has

– zero or an even number of nitrogen atoms, its
molecular ion will appear as a even m/z value
– an odd number of nitrogen atoms, its molecular
ion will appear as an odd m/z value
B. Mass Spectroscopy:
Following the pioneering work of Schumann and coll.
[78] and Neuner-Jehle and coll. [79 , 80] who worked
out the principal fragmentation patterns of the most
important QA , mass spectrometry (MS) developed
into a convenient method of QA characterisation [81] .
Mass spectrometry is widely used today, since QA
usually provide distinctive fragmentation patterns in
the electron impact mode (EI-MS).
Chemical ionisation (CI-MS) , field desorption (FD-
MS) and fast atom bombardment (FAB-MS) are
suitable for identifying molecular ions of QA esters
and tricyclic alkaloids , whose molecular ions are
usually obscure or absent in EI-MS spectra .
A major advantage of MS is the possibility it gives of
combining the high resolution power of capillary GC
with sensivity and informativity of EI- or CI-MS.
Work using GC-MS was very much facilitated after

1980 by the development of new GC capillary
columns, the development of new methods to position
the GC column outflow near the MS ion source and,
most importantly, by the improved possibilities of data
processing.
During a GC-MS run, MS spectra are recorded

continuously, thus a typical GC-MS run consists of up
to 1000 spectra.
These data can easily be processed by computer, with
background subtraction or the profiling of individual and
charateristic fragment ions (‗ion chromatography ‘), which are
helpful to select the relevant MS spectra.
A normal plant extract contains a number of hydrocarbons and

contaminants, such as phthalic acid derivatives, which can
be identified because of their distinctive m/z 149 fragment
[82].
In Table I-4 the molecular ion and the four most informative
fragments and their abundances are tabulated.
For CI-MS data the molecular ion (M + H+) is given. The data
for FAB-MS were reported, among others, [41]. Field
desorption and chemical ionisation techniques are also
helpful in the direct analysis of alkaloid mixtures [83].
Laser desorption mass spectrometry (LAMMA 1000) is
a soft ionisation method which produces significant (M
–H) + ions and informative fragment ions.
This method has been used to localise the site of QA
storage in plant tissue; sparteine could be located in
the epidermal and subepidermal cell layer of Cytisus
scoparius stems. Lupanine is accumulated in the
epidermis of petioles of Lupinus polyphyllus [84].
Alkaloid extracts of many legumes contain piperidine
alkaloids such as ammodendrine, N–methyl ammo
dendrine, hystrine or smipine. These alkaloids also
derive biogenetically from lysine via cadaverine [85].
Mass spectral information is provided in Table I-4.
ANALISIS ALKALOID
Contoh: analisis alkaloid quinolizidin dalam madu (HPLC-APCI-MS)
ANALISIS ALKALOID
Contoh: analisis alkaloid ergot pada Festuca arundinaceae (HPLC-ESI-MS)
Nicotine
Nicotine
Caffeine
ANALISIS ALKALOID
Contoh: analisis alkaloid quinolizidin dalam madu (HPLC-APCI-MS)
ANALISIS ALKALOID
Contoh: analisis alkaloid ergot pada Festuca arundinaceae (HPLC-ESI-MS)
NMR Signals
• The number of signals shows how many
different kinds of protons are present.
• The location of the signals shows how
shielded or deshielded the proton is.
• The intensity of the signal shows the
number of protons of that type.
• Signal splitting shows the number of
protons on adjacent atoms. =>
Spektrometri NMR
I. Letak puncak (geseran kimia)  d
• Menunjukkan jenis gugus fungsi dan ikatan
II. Tetapan gandeng (Coupling constant)  J

• Menunjukkan posisi H diantara H tetangganya
III. Integrasi
• Menunjukkan jumlah H dan C
IV. Korelasi geseran kimia dengan H dan C

• Menunjukkan fragmen struktur
V. Penetapan Kerangka molekul

• Beberapa golongan senyawa dapat ditetapkan
kerangkanya dari data spektrum NMR
NUCLEAR MAGNETIC RESONANCE
Nuclear Magnetic resonance:
– Permits the establishment of the structural
skeleton of the compound investigated.
– 1HNMR showed resonances of protons while
13C NMR showed the C resonances.
– Able to establish the connectivity between

carbons and protons.
– One dimensional and two dimensional
techniques available:
• COSY, HMQC, HMBC, NOESY etc.
– For 1HNMR ~ 1-5 mg (pure) sufficient
– For 13C NMR ~ 20 mg sufficient.
Chemical Shift of
Solvent
H2O (or HOD)
Acetone 2.8
Acetonitrile 2.1
Benzene 0.4
Chloroform 1.6
Dimethyl Sulfoxide 3.3
Methanol 4.8
Methylene Chloride 1.5
Pyridine 4.9
Water (D2O) 4.8
1H NMR Chemical 13C NMR Chemical
Solvent
Shift Shift
Acetic Acid 11.65 (1) , 2.04 (5) 179.0 (1) , 20.0 (7)
Acetone 2.05 (5) 206.7 (13) , 29.9 (7)
Acetonitrile 1.94 (5) 118.7 (1) , 1.39 (7)
Benzene 7.16 (1) 128.4 (3)
Chloroform 7.26 (1) 77.2 (3)
Dimethyl Sulfoxide 2.50 (5) 39.5 (7)
Methanol 4.87 (1) , 3.31 (5) 49.1 (7)
Methylene Chloride 5.32 (3) 54.00 (5)
Pyridine 8.74 (1) , 7.58 (1) , 7.22 (1) 150.3 (1) , 135.9 (3) , 123.9 (5)
Water (D2O) 4.8
Nuclear Magnetic Resonance Spectroscopy
1H NMR—Chemical Shift Values
The standard for protons is the proton resonance in TMS, Si(CH3)4. Other
references are used for other nuclei. The table below shows typical ranges of
chemical shifts for protons in different groups.
O-H and N-H Signals
• Chemical shift depends on concentration.

• Hydrogen bonding in concentrated
solutions deshield the protons, so signal
is around d = 3.5 for N-H and d = 4.5 for
O-H.
• Proton exchanges between the molecules
broaden the peak.
=>
Hydrogen and Carbon
Chemical Shifts
=>
CH, CH2, CH3
Nicotine
CH, CH2, CH3
Aromatik, NH
Caffeine
NCH3
Aromatik
OCH3 HO 3 4
Aromatik 2 5
8 3 10-OMe 1-OMe
1
NH
11 H3CO
H
7
11
8
H3CO 10
9
OH
norboldine
7-OMe
12-OMe 2-NMe -OCH3, -NCH3

7‘-OMe
Aromatik, NH 6-OMe
2‘-NMe
6‘-OMe
8 8‘
13 11‘ 5‘
14‘ 13‘ 5
10‘
14‘
10
11
4 5 5' 4'
OCH 3 H3CO 6'
O-methyldauricine 3
6
3'
7'
N N
H3C 7 OCH 3 H3CO CH3
1 8 8' 1'
10 10'
9 11
11'
9'
12
O
14 14'
12'
13
OCH 3
13'
5,0 4,5 4,0 3,5 3,0 2,5 2,0 1,5 1,0 0,5 0
H-2‗‘ H-
6‘‘
H-1‗‘
CH, CH2, CH3
Aromatik
10 – 60
Nicotine
H2CNH CH2
60–110
110–170
Nicotine
CH, CH2, CH3
Caffeine 10 – 60
Aromatik
110–170
Caffeine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Nicotine
Caffeine
Caffeine
Caffeine
Caffeine
Caffeine
Caffeine
Caffeine
Caffeine
160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10
Spektrum 13C-NMR Tinocrisposid [1]
150 100 50 10
4
OH
3
3 4 HO 2 5 2 3 4
H3CO 2 H3CO H3CO 2
5 1 5
3
1
1 NH 1
N H3CO 4 10 N CH3
H3CO H H3CO 6a
16
7 7 15 9 H
11 7
11 O 5 12
N
8
10 8 8 6 H H 11
H3CO 10 8
9 9
9 H3CO 7
OH O O 10
lysicamine norboldine pallidinine tetrahydropronuciferine
3 4
H3CO 2
5
5 4
1
NCH3 H3CO
3
4 5 5' 4' H3CO 6
OCH3 H3CO 6' H
3 3'
6 12 7 N
7' HO 7 CH3
N N 8 1
7
8
H3C 1 OCH3 H3CO 1' CH3 11
8 8' 9 11
10 10'
10 HO 9
9 11
HN
11' NH 12 10
9' 7' 2'
12
O
14
12' 14' 13
15
13
OCH3 6' 3' H3CO 14
13'
4'
5'
CH3
reticuline
O-methyldauricine
phoebescortechiniine
2-OMe
1-OMe
11 5
8 4 9 10
lysicamine 3 4
H3CO 2
5
1
N
H3CO
7
11 O
10 8
9
10-OMe 1-OMe
8 3
11
3 4
HO 2 5
1
NH
H3CO
H
7
norboldine 11
8
H3CO 10
9
OH
7-OMe
12-OMe 2-NMe
7‘-OMe
6-OMe
2‘-NMe
6‘-OMe
8 8‘
13 11‘ 5‘
14‘ 13‘ 5
10‘
14‘
10
11
O-methyldauricine 4 5 5' 4'

OCH 3 H3CO 6'
3 3'
6
7'
N N
H3C 7 OCH 3 H3CO CH3
1 8 8' 1'
10 10'
9 11
11'
9'
12
O
14 14'
12'
13
OCH 3
13'
3-OMe 6-OMe
-NMe
4 1
5
16
10ax 9
15
8 10ef
OH
2
H3CO
1
3
pallidinine
4 10
16
15 9
5 N
6 H H
8
H3CO 7
O
4‘-OMe
2-OMe -NMe
1-OMe
2‘
5‘
2‘ 3‘
1‘-NH
3 4
H3CO 2
5
1
NCH 3
H3CO
phoebescortechiniine H
12 7
8
11
10 9
HN
NH
7' 2'
6' 3'
4'
5'
CH3
reticuline H3CO
5 4
3
6
N
HO 7 CH3
8 1
11
HO 9
12 10
13
15
H3CO 14
-NMe
2-OMe
1-OMe
7
6a 12
tetrahydropronuciferine H3CO
3 4
2
5
1
N CH3
H3CO 6a
H
7
12
8
11
9
O 10
PENENTUAN STRUKTUR ALKALOID: spektrum NMR
3.85
3.82
6.63 -OCH3
-OCH3
6.59
7.02 6.65
H3CO
NH
H3CO
3.26 2.98 2.89
HO
4.19
2H 4H 1H 6H 1H 3H 2H
7.0 6.0 4.0 3.0

5.0
ppm

Hand Out Fitokimia Alkaloid 16 Des 2016

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Hand Out Fitokimia Alkaloid 16 Des 2016

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1.

Ciri umum/ kerangka utama

This was the first

Found only in the Opium Poppy - Papaver somniferum

Rauwolfia serpentina (L) Benth.

derives from the word alkaline;

Definition: the term ―alkaloid‖ (alkali-like) is

• Glycosidal form e.g. Solanine in Solanum.

Oxycodone Oxycodone HCl

Alkaloid bebas/ basa Garam alkaloid

- Larut dalam pelarut - Larut dalam air

Alkaloids are cyclic organic

• Tersebar pada ~ 20% tumbuhan tingkat tinggi

Many ideas have been advanced:

• Turunan ornitina  atropina

NH2 NH2 NH2

Methylations could also N CH3

basic due to the contains nitrogen

This was the first

Found only in the Opium Poppy - Papaver somniferum

Residu Ekstrak CHCl3/NH3

• partisi dengan lar. 5% H2SO4

Lapisan air Lapisan CHCl3

2) Alkaloidal coloring reagents:

3) Special tests for alkaloids:

• Salts are usually soluble in water and, insoluble or

• Weak bases e.g. Caffeine

• Most alkaloids contain Oxygen and are

• Some alkaloids are free from Oxygen and

• Reaction with acids:

Ergot Alkaloids Lumi Alkaloids

Ekstraksi eter minyak Ampas

Ekstrak EtOAc Lapisan asam tartrat 2%

Ekstrak EtOAc Lapisan air basa

Ekstraksi CHCl3 Lapisan air

• Basic Chemical Skeleton

True alkaloids derive from amino acid and they

These alkaloids are highly reactive substances with

All true alkaloids have a bitter taste and appear as

Moreover, most of them are well-defined

True alkaloids may occur in plants

pyrrolidine pyrrole piperidine pyridine

Protoalkaloids are compounds, in which

Such kinds of alkaloid include compounds

C6 H5 CH3 H3CO NH2

Alkaloids without heterocyclic nitrogen atoms

Pseudoalkaloids are compounds, the basic carbon

In reality, pseudoalkaloids are connected with amino

They are derived from the precursors or postcursors

These alkaloids can also be derived from

The N atom is inserted into the molecule at a

Common classification schemes :

(1) Biosynthetic Origin:

Depends on types of precursors or building block

• Turunan ornitina  atropina

NH2 NH2 NH2

Methylations could also N CH3

The aconite alkaloids are derived from the aconitum

Depends on the type of heterocyclic ring systems

It is usual to classify alkaloids according to the

pyrrolidine pyrrole piperidine pyridine

quinolizidine pyrrolizidine tropane

benzylisoquinoline purine -phenylethylamine