A. W.

Sangstor
University College
(1 Determination of Alkaloid Structures

I
of the West lndies I. Isolation, characterization, a n d
Jamaica
physical methods

There are several well-known books on small. The Gymnosperms and Pteridophytes are poorly
the chemistry of the alkaloids (I), but these tend to represented and so far as is known alkaloids are virtually
discuss the detailed chemistry of particular groups of absent from the lower groups of plants with the excep
alkaloids. It is the purpose of these articles to outline tion of one or two families of the fungi (e.g., ergot) (8).
the general methods of approach in alkaloid chemistry.' Alkaloids generally occur in the plant tissue a t points
The term "alkaloid" (alkali-like) is applied to the of intense cell activity such as the leaves, roots, bark,
large group of basic nitrogen-containing natural prod- and seeds and are generally regarded as plant met,abolic .
ucts of vegetable origin. They are usually colorless byproducts. There is considerable variation in the
(berberine is yellow), well-crystalline solids, but some alkaloidal content in the plant depending on such factors
from the tobacco, hemlock, and pomegranate root bark as time of reaping and methods of cultivation and selec-
are liquids,. The bases are usually optically active tion.
(mainly levorotatory) though a few such as coniine, A knowledge of the botanical origin of an alkaloid
cinchonine, laudanosine, pilocarpine, and pelletierine can be useful in giving indications of the structural type
are dextrorotatory,andothers like berberine,papaverine, of alkaloid which is likely to be present. Plants of
and pilocereine are optically inactive. I n most cases the same family generally tend to produce alkaloids of
the nitrogen atom forms an integral part of a cyclic the same structural type. This is a generalization which
structure. Classifications of alkaloids usually exclude has to be applied with caution for there are alkaloids
simple amines, purines, hetaines derived from amino such as nicotine and berberine which are found in
acids of proteins, choliie, and other bases of biological several different plant families. Also plants such as the
importance which in contradistinction to the typical opium poppy give a number of different structural
alkaloids are not limited to one or a few species of types of alkaloids though the latter are probably bio-
plants. genetically related.
Alkaloids occur most abundantly in the higher orders
of the plant kingdom and have been found in over 90 Alkaloids occur usually as salts of the common plant
different families of flowering plants (2). acids. I n certain plants, however, a specific acid may
The majority of these families belong to dicotyl- be found associated with certain alkaloids such as
edenous orders and the number of monocotyleden~us quinic acid in the cinchona group, aconitic acid in the
families showing the presence of alkaloids is relatively aconitines, and meconic acid in the opium group. A
few alkaloids such as narceine and narcotine are found
The present article deals with methods of isohtion and char- free in nature, probably because of their low basicity.
acterization and the applications of physics1 methods to the de- A few alkaloids as found in the solanum and veratrum
termination of alkaloid structures. The second paper, to be
published in October, will describe chemical methods of groups occur as glycosides of such sugars as glucose,
.structural determination. rhamnose, and galactose. Other alkaloids as in the

454 / Journal of Chemical Education

 Table 1. The Occurrence and Use of Some Common Alkaloids

Alkaloid Source Use Alkaloid Source Use

Arecoline Betel nuts Masticatory; vermifuge Morphine Opium Pain relief. narcotic
Atropine Belladonna Mydriatic in ophthalmalogy Nicotine Tobacco Stimulant: insecticide
Caffeine" Coffee, ten Stimulant Papaverine Opium Spasmolytio drug
Cocaine Coca. leaf Local anaesthetic Physo~tigmine Calabar bean Gastrointc~tinel and bladder
Codeine Opium Pain reliever stimulant
Colchicine Crocus Plant genetics Pilocarpine Jaborandi Diaphoretic (sweat producing)
Cusparine Angustura bark Stimulant Quininr Cinchona bark Malaria fever
Emetine Ipecac roots Emetic; amoebic dysentery Reserpine Rauwolfia Tranquilizer
Ephedri?e "Ma Huang" Asthma Scopolamine- 1)atura Narcotic
Ergonomne Fgot Obstetrics, oxytocic Sparteine Broom Cardiac treatment
Febrifugine Ch'ang Shan" ~nti-ma1a;ial Strychnine Strvchnos Pest exterminator
Mescaline "Mescal buttons" Hallucinatory principle Tubocurarine Chondrodendron Nerve paralysis
Yohimhine Yohimbe bark Aphrodisiac (sex stimulation)
a Sometimes classed as an alkaloid cf. (Ic).

tropane, aconitine, senecio, veratrum, and yohimbine tion is made basic and again extracted with an organic
groups are encountered as esters of various organic solvent usually ether or chloroform to give a crude mix-
acids of varying complexity. A few others occur as ture of bases. Some alkaloid hydrochlorides are soluble
amides such as piperine and the ergot alkaloids. in chloroform and this property can be used for purposes
The study of the chemistry of the alkaloids has been of separation (5b). A modification of the above process
particularly att,ractive to chemists partly on account is the extraction of the plant material with an aqueous-
of their wide occurrence and diversity of physiological organic solvent mixture and subsequent separation and
action on the animal organism and partly on account drying of the organic layer. Non-basic alkaloids can
of the complex structural and synthetic problems in- usually be obtained after removal of any acid or alka-
volved in the elucidation of their structure. Many of line constituents of the organic layer.
these investigations have been classics in the realms of Acid extraction, though less frequently used, has
structure elucidation and synthesis. The occurrence been applied with success to the isolation of alkaloids
and uses of some of the commoner alkaloids are shown from undried plant material in the field using ion ex-
in Table 1. change techniques. Alternatively the dried plant
material is extracted with a dilute acid such as sulfuric
Isolation Methods or tartaric. Tartaric acid has the advantage of tending
to give a purer product. Where there is danger of
The presence of alkaloids in plants is first established decomposition of the plant base, the extraction is done
by tests on extracts from the plant with various alka- in the cold. The aqueous solution is hasified and either
loidal reagents (Sa). The presence of a precipitate or extracted with an organic solvent or steam-distilled to
turbidity with such common reagents as Mayer's remove volatile components. Quaternary alkaloids
(potassium mercuric iodide), Wagner's (iodine-potas- which are usually quite soluble in water are generally
sium iodide), Dragendofi's (potassium bismuth iodide), recovered by precipitation with mercuric chloride or
Sonnenschein's (phosphomolybdic acid), and Scheibler's Mayer's reagent and the resulting mercury complex de-
(phosphotungstic arid) indicates the presenre of alka- composed with hydrogen sulfide. Some alkaloids which
loids. occur as N-oxides are quite soluble in water. They
"Mayer'e resgmt is most commonly used as it is generally the may be obtained by reduction with acid zinc dust fol-
most sensitive. About 1 g of plant material is digested with lowed by chloroform extraction. These methods are
5-10 ml of dilute HCI and the filtrate tested with a. few drops summarized in Table 2 with appropriate examples.
of the reagent. A creamy precipitate or turbidity gmcrally
indicates the- presence of alkaloids." Separation and Purification Methods
The extraction of the total alkaloids from the plant, In the early days of alkaloid chemistry the separation
material is generally accomplished either by extraction of mixtures of alkaloids often involved tedious frac-
with acids or with organic solvents. Various modifi- tional rrystallizations or precipitations. Nevertheless,
cations have been worked out empirically for the ex- excellent separations were achieved. However the
traction of particular alkaloids. Solvent extraction is complexity of some alkaloid mixtures is such that for
usually the more common procedure. The dried plant. a time only the more abundant components could be
material is usually subjected to a preliminary extraction examined. The advent of more elegant techniques of
with light petroleum to remove plant oils. Most alka- separation such as counter-current distribution, ad-
loids and their salts are insoluble in petroleum. The sorption, and partition chromatography have made pos-
alkaloid bases are then liberated from the salts present sible the study of many of the minor components (10).
in the plant material by treatment with ba.se (calcium Ion exchange, though potentially a good method, has
hydroxide, ammonium hydroxide, or sodium carbonate) had little application in separation. These techniques
and then extracted with a suitable organic solvent surh are too well known to require elaboration. A summary
as chloroform, ether, benzene, ethanol, methanol, or of the main methods of separation with examples of
methylene chloride. The alkaloids are then extracted their application is given in Table 3 and Figures 1and 2.
with acid from the organic solvent or pasty residue that Some criterion of the effectiveness of a separation is
remains after removal of the solvent. The acid solu- required. Paper chromatography and to a lesser ex-
Volume 37, Number 9, September 1960 / 455
 Table 2. Examples of Alkaloid Isolation Methods
Acid Extraction Methods Solvent.k>xtraction Methods

Cyclic Percolation of dried plant material with Extraction Extraction Addition of

extraction dilute acid, or hot extraction base and ex-
of undried tractor with
plant and organic sol-
absorption
on ion
exchange
lcolumn ( 4 ) 1 I

Concentration and basification I I Separation

of solvent
Precipitation Acidification
with alcohol of quaternary

%-
& removal of
and ammonia alkaloids a i t b loils with I
mercuric chloride components petroleum

Removal of and extrac-

tion with

'i
distillate solvent organic
solvent
of base by from ~olution
extraction

I Crude Alkaloids

tent paper electrophoresis have been useful in this con- Structural Investigation b y Physical M e t h o d s
nection (Table 3, Fig. 3).
The individual alkaloid is characterized by constancy Physical methods are becoming increasingly impor-
of its melting point, boiling point, optical rotation, tant in the structural elucidation of chemical com-
ultraviolet spectra, infrared spectra, and any other suit- pounds. A few examples of each method must suffice
able physical properties, and also by the preparation to illustrate the approach. It should also be em-
and characterization of derivatives and salts. Often phasized that physical and chemical methods go hand
the purification of an alkaloid through one of its salts in hand in structural investigation.
is more convenient; a useful test for homogeneity is Ultraviolet Spectra
the inter-convertability of the alkaloid and its salts.
Quantitative methods of estimation of alkaloids which UV spectra have provided the keys to the solution
are generally either colorimetric, volumetric, or gravi- of many problems in alkaloid chemistry. W spectra
metric depend largely on the types of functional groups may he useful in pointing to the type of structure
present (Sd). likely to be found. Broadly speaking the spectra of
alkaloids may be divided into three main classes (26).
No UV absorption in the range 200-1000 m#. Examples are
the saturated ring systems such as the pyrrolidine and piperi-
dine alkaloids.
Simple aromatic chromophoric systems such as substituted
benzene or pyridine with medium intensity absorption a t rerelsr
t i d y short wavelengths (230-300 mr),
Alkaloids with complex ehromophores, either aromatic
nuclei in conjugation vith other chromophores or fused aro-
matic systems.
Examples of the use of spectral methods in structure
determination are seen with the alkaloids sempervirine
(27), strychnine ($a), and emetine (29).
The use of UV spectra in the assignment of structures
is illustrated by the following example from the mor-
phine series. The structure of the two dihydro the-
baines was corrected by the use of UV spectra (30).
The structure of the lithium aluminum hydride re-
duction product was shown to be I1 since the extinction
ELUATEFRACTIONS values indicated a conjugated system. The structure
Figure 1 . Partition chromatographic seporotion of pomegranate root of the sodium and ethanol reduction product was shown
bork d k d o i d s [by permission from the J. Phorm. and Phormocol.) 1141. to be I11 since the extinction value showed a marked
456 / Journal of Chernicol Education
lowering indicating the absence of a conjugated double Table 3. Alkaloid Separation Methods
bond system. General
Example of Refer- refer-
Method rtnnlicat,ion ence ence
Fractional
crystallisation
Quinine alkaloids . .
(11)
Fractional Hemlock alkaloids Xe)
distillation
Senamtion bv Verrttrum alkaloids 12
ksction
Adsorption
- by acetylation
Senecio alkaloids 15 19, 20
chromatography
Partition Pomegranate root 14 19
chromatography bark alkaloids (Fig. 1)
Counter-current Veratrum alkaloids 15 dl(a, b )
distribution (Fig. 2)
Ion exchange Yohimbine alkaloids 16 W
Paver Curare alkaloids 17 19. 25
Ehromatography (Fig. 3)
Paper electrophoresis Stryohnos alkaloids 18 B , 2 4 25

I. Thehaine 11. LiA1H4product 111. Na/EtOHproduct

r = 6500 r = 11,000 r = 2000 sewations could be reconciled by a structure VI for
A = 283 mp X = 284 mp A = 282 mp
the intermediate (96).
The UV fluorescence spectra of some alkaloids have
also been determined (31).
Infrared Spectra
Infrared spectra have been extensively used in the
elucidation of the structures of organic molecules
(9% 99, 94).
Some of the commoner functions found in alkaloids Basic Sfrengtl~sand Polarogmphic Measurements
and their characteristic infrared absorption frequencies The determination of pK values has become increas-
are shown in Table 4. The infrared spectra of alkaloids ingly important in structure determination and par-
have been used for comparative purposes especially in ticularly in the elucidation of structural details. An
the fields of drugs and narcotics (95), and also in fol-
example of this application is in the placing of the
lowing the course of synthetic and degradative ex- double bond in the structure of retronecine, VII, the
periments. alkanolamiue hydrolytic cleavage product of the al-
Thus the infrared spectra were used to reformulate kaloid monocrotaline (4ia). The a-p or 8-7 position-
the course of the synthesis of N-methylisomorphinane,
ing of the double bond relative to the nitrogen atom
IV. One of the intermediates in the synthesis was the was correctly postulated on the basis of pK measure-
postulated nitrile, V. This was shown to be incorrect
ments and subsequently confirmed by other methods.
as the infrared spectrum showed no nitrile bands a t
Retronecine is a weaker base than the corresponding
2240 cm-', but showed bands at 3400 em-', (secondary saturated amine platynecine, VIII. This is compatible
amine), 1664 em-' (cyclic amide), 1676 cm-', and 1692
with a p-y unsaturation whereasau w-8 type of unsatura-
em-' (cyclic conjugated carbonyl). The latter ob- tion would lead to an increase in basic strength due to
the facile transformation:

-N-C=C-
'
I

A
+ H+ - +
-N=C-
'
H
A-
1
H

VIII
pK, 10.22

The application of pK values to the steric arrange-

ment in some terpenoid alkaloids has been discussed
(43).
The presence of a tropolone ring in colchicine was
postulated on the basis of the similarities in the polaro-
grams of colchicine and ythujaplicin (49).
X-Ray Crystallography
No.E
TU~ The application of X-ray crystallographic methods
Figure 2. Twenty-four-tube counter-current distribution of veratrum
to the complex three-dimensional structures often en-
alkaloids (by permission from Anal. Chem.) 175). countered in alkaloid studies has been very rewarding
Volume 37, Number 9, September 1960 / 457
 Nuclear Magnetic Resononce Spectro
The increasing interest in NMR spectra lies in the
fact that many details of structure can be decided
unambiguously by this method though this may not
be possible using ultraviolet or infrared spectra.
Also simple model compounds can be utilized to obtain
a "fingerprint" of the portion of the spectrum under
consideration. This is analogous to the use of model
compounds in UV and I R work but in some cases the
model suitable for NMR comparison may be more
readily available.
An example of the application of the method in the
complete proof of the structure of the alkaloid Luna-
crine, XII, is outlined below.
0

XI1
I I I I I
The structure was elucidated by a detailed analysis
I 6 1 4 1 T- of seven well defined groups of lines in the NMR
Figure 3. Paper chrornotogrorn of curare alkaloids (from Helv. Chim.
A < f d (17).
spectrum which were assigned as follows (49): (a)
the hydrogen atom at position 5 of the quinolone system;
(b) the aromatic hydrogen atoms at 6 and 7; (c) the a-
(41b). Classical examples of the application are the hydrogen atom of the dihydrofurano ring; (d) the
studies of the alkaloids strychnine and colchicine.
Thus the X-ray spectral method made it possible to Table 4. Some Infrared Absorption Characteristics of
distinguish between the structures I X and X for the Alkaloids
alkaloid colchicine and showed that the structure X was
Infrared absorption om-1
the correct one. S = Strong
Intensity: M = Medium Refer-
NHCOCH, Group W = Weak ence
-0H- 36253540 ( S ) non-associated 3b, 37
3400-3200 (S) associated
Phenolic acetate 1767-1764(S) 38
Alcoholic acetate 1742-1735 (S)
C=O 1850-162O(S) 37
IX 0 C=N- 1660-1480 (S) 3s
x OCHs
C=C 1680-1620 ( S M I 32
Recently the structure of the complex alkaloid gelse- near 3030 (W-M-S) Sb, 37
mine, XI, has been independently postulated by both 1640-1600 (S)
near 1500 (W-M-S)
chemical and X-ray methods (44).
32
..
3480-3460 ( 8 ) indale 37
\NH 3480-3440 (S) 2' amines
/
3280-3200
3380-3205
(S) associated
(S) 2nd amine 1 39
\$/
Rotation01 Evidence
Ammonium 244k2350 (W)
/ H' 40
Optical rotational evidence has been used in a few c3
cases to determine stereochemical configurations. Thus =N-
Immonium 2100-1980 (W)
rotational evidence was used to establish stereochemical
correlations between yohimbine and known steroid
4
NHsf2" Aminesalt 1620-1560 (S-M) deformation
structures (45), and the absolute configuration of eme- mode
tine has been deduced from optical rotational differences Hydrochloride salts 3000-2500 (M-W) Series of bands Sb
(46).
Optical rotatory dispersion has had limited applica- methoxyl- and N-methyl-hydrogen atoms; (e) the
tion in the field of alkaloids, largely due to lack of suit- &hydrogen atoms of the dihydrofurano ring; (f) the
able instruments, though some correlations between tertiary hydrogen atom of the isopropyl group; and
the diterpenoid alkaloids and the diterpenes have been (g) the C-methyl-hydrogen atoms.
made (471, and recently the absolute configuration of Other examples of the application of NMR spectra
emetine has been independently determined with the in structure determination are illustrated by studies on
aid of optical rotatory dispersion curves (48). the alkaloids lycodine (50a) and gelsemine (506).
458 / Journal o f Chemical Education
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Volume 37, Number 9, September 1960 / 459