STUDIES ON HASHISH

I. ISOLATION & IDENTIFICATION OF CANNABINOLS AND

EFFECT OF CERTAIN FACTORS

Z. I. EL-DARAWY, M. I. ROUSHDY*, A. M. RIZK*,

F. M. HAMMOUDA* & Z. M. MOBARAK

National Centre o f Social and Criminological Research,

A wkaf City, Gezira Post Office, Cairo, Egypt

ABSTRACT

Fractionation of the Hashish constituents as well as the isolation of certain consti-

tuents on a preparative scale was carried out using several physio-chemical techniques
viz. solvent fractionation, column and thin-layer chromatographic methods. Six con-
stituents viz. cannabidiolic acid, cannabidiol, cannabinot, tetrahydrocannabinol, can-
nabichromene and a positive Beam component (cannabidiol isomer) were isolated and
identified using chromatographic data and ultraviolet spectrophotometry. Separation
of the Hashish constituents was also achieved by gas-liquid chromatography. The effect
of certain factors viz. silica gel, acidity and alkalinity was studied revealing a remark-
able change.

Cannabis or Hashish, one of the oldest drug plants, represents at the time
being an important group o f unique character in the modern growing fields
of socio- and psychopharmacology (Chopra & Chopra, 1957). The abuse of
this drug was, and still is, more wide spread throughout the World than the
abuse of any other single drug known.
Through the centuries many chemical races of the plant have come to
existance and are progressively recognized. However, the botanical origin so
far seems to be namely Cannabis sativa L. and the active drug remains to be
the oleo-resin obtained from the flowering tops of the unfertilized female
plant. Botanically there is only one species of Hemp namely Cannabis sativa
L.; such names as C indica (Hamilton, 1913), C satipa var. indica (Merrill,
1938), C americana (Merrill, 1938; Wolf, 1949) are not botanically admis-
sible and must be regarded as ' c o m m o n ' names o f the same calibre as Indian
Hemp and Hashish.
Several vernacular names viz. Hashish, Magoun, Kif, Bangi, M a r i h u a n a . . .
etc. (Watt & Breyer-Brandwijk, 1962) are known for Cannabis in the dif-

* National Research Centre, Dokki, Cairo.

Qual. Plant. Mater. Veg. XXI, 4 : 311 - 325, 1972 311

ferent countries. The numerous preparations having Hashish activity are all
obtained from the Cannabinaceae famiiy.
The classical constituents of Cannabis are cannabidiol (I), cannabinol (II)
and tetrahydrocannabinol (III, IV). The structure of these compounds has
been confirmed by the intensive work of several authors (Cahn,
1930-1933; Bergel et al., 1943; Adams et al., 1949). Cannabidiolic acid (V)
as well as another acids were also isolated from the leaves and stem tops
(Krejci & Santavy, 1955; Krejci et al., 1958, 1959; Schultz & Haffner, 1958,
1959). Several other constituents e.g. cannabinolic acid, cannabigerol, can-
nabichromene.., etc (VI-XVI) had been isolated from Hashish (Claussen
et al., 1966, 1968; Goani & Mechoulam, 1964, 1966; Hilvely et al., 1966;
Kane, 1968; Korte et al., 1965; Mechoulam et al., 1964-t969; Obata,
1966; Okamoto, 1967; Shoyama et al., 1968, i970; Spulak et al., 1968;
Yamauchi et al., 1967, 1968; Vollner et al., 1969).
Thin-layer chromatography had been successively applied by several
authors for the separation of cannabinols. The first application was that by
Krejci (1961). Different adsorbents viz. silica gel (impregnated with
dimethylformamide or AgNO3 or without) (Korte & Sieper, 1964 a,b; Turk

CH:j C.H~

H3c I1 6H ~ -CsH~ H~C I

C5 H II
CH2 CH3
Cannabidiol(I) Cannablnol (~)

CH3 CH3

H3C C5Hll H3C CsH11

CH3
z~9- ti- an s
Tetr ahydr o- ~B- t rans Tetrahydro-
cannabinol (TIT) c annabinol ( ] ~ )

C,H3 CH 3

O~COOH O~COOH

HC--C H3C- - ~ ' 0 ~ C 5 H l t

CH2 CH3
Cannab(d(ol;c ac;d Cannabinot(c acid
(Y} ('£I)

312
et al., 1969 a,b), alumina (Stone, 1969) or silica gel and poly (vinyl
alcohol) dipped in Et2NH (Grlic, 1970) were used. The chemical detection
of the chromatographed spots, which is far more superior to physical
detection was carried out by spraying with a variety of reagents viz. Gibbs
(1927), Beam (1911), Duquenois (1938) and tetrazotised o-dianisidine
(Fietz-David & Blangey, 1949).
Although several methods for separation of the cannabinols by gas-liquid
chromatography, have been described in the literature, all have had certain
limitations. No success was obtained with the relatively non-polar columns
of methyl, silicone gum (Claussen et al., 1966; Heaysman et al., 1967;
Okamoto, 1967) and cyanosilicone gum (Caddy et al., 1967); while the
somewhat polar stationary phase (Carbowax 20 M) (Heaysman et al., 1967)
gave better separation of the cannabinols. On the other hand, the high
temperature 230°C required for tile column is liable to limit the efficient
life of the column. The best resolution obtained by Stone (1969) using
neopentyl glycol adipate plus trimer acid column, still needs high column
temperature of 220°C.
c H3 CH3

O~ COOH O~j~ ooH

,,c..;. )..IL
i- O- v ~CBHI 1 H3C" "O" ~ "CsHI1
C H3
~9- ¢ran s -Tetrahydro- ~ Tetrahydro -
cannablno[ic acid ( ~ ) cannablnolic acid A
(~n-r)
C.H3

H3O C ~ C5Hll

~c" -o- y -%H.

COOH
A1-Tetrahydro- Can~ab;gerol
cannablno{ic acid B (X)
( T'X"}

CH2= C.CH 3 (CH2~3C.CH3=CH

HO" ~ C s H ~i

Cannabig• rol monomethyi

ether ( ~ )
Cannabigero|ie
acid ( ~ )

313
 OH OH

C,H2-CH csH, c %~2c °C,.CH2.CHrI..0/%j-.% H1,

CH CH3 CH3
II
C..,
C'~3 CH 3 Cannabichromenlc acid

Cannablchromene t ~ )

CH3

H%lC5 H2

H3C 02C 0
~CsHII
xk .J~ CH3 H OH
H11C5~ ~ ~O ~ \
CH3 Cannabipinot
Tetr ahydr o c annabitr iot OR
CannabldioI c a r b o x y t i c acid Cannabicy¢toi
ester (~!~'1 ( ~ )

Adulteration of Hashish by mixing with different herbs or drugs, makes

its detection and identification somewhat difficult. This work deals with the
fractionation of Hashish constituents using several physico-chemical tech-
niques involving solvent fractionation, thin-layer and column chromato-
graphy of certain constituents on a preparative scale.

EXPERIMENTAL AND RESULTS

Preparation of Hashish Resin

10 g of powdered Hashish were macerated twice with methanol (25 ml

each) for 24 h, in dark, at room temp. Methanol was evaporated in vacuo at
40°C and the residue was taken with petroleum ether (40-60°C) - ether
(80:20) and filtered from the insoluble matter. The filtrate was passed
through a column packed with silica gel and elution was carried out using
the same solvent mixture.

Thin-Layer Chromatography

Plates coated with silica gel G, using different solvents were tried. The
solvent petroleum ether (40-60°C) - ether - ethyl acetate (90:5:5) proved
to be the best revealing the presence of 11 components, upon spraying with
tetrazotised o-tolidine (Fig. 1).

314
 & & c,o
THC
C~ CBN
CBC

0 0 :BO"

CBDA

t 2
Fig. ( t )
Thin-layer Chromatogram of Purified
Hashish Re~in
Adsorbent : S i l i c a gel G.
Solvent s~'stem ; Pet, ether ( 4 0 - 6 0 ' C ) -
Ether-Ethyl acetate (go : 5 : 5)
Sgraying reagent : 1) Beam,
21Tetrazotised o-totidine.

Isolation o f Cannabidiolic A cid

Cannabidiolic acid (CBDA) was separated from the purified Hashish resin
as follows: The petroleum ether - ether etuate was shaken with three
successive portions (20 ml each) of aqueous solution of sodium carbonate
(5%) and sodium sulphite (5%) (I : 1). The combined aqueous solution, after
washing with petroleum ether-ether was rendered acidic with H2 SO4 (3%)
and the liberated CBDA was extracted with ether. The ether extract was
dehydrated over anhydrous Na2 SO4 and the solvent was removed in vacuo.
TLC revealed the presence of CBDA which was further purified by prepara-
tive TLC using petroleum ether (100-140°C) - acetone (1:1). The isolated
CBDA was found to possess the same Rf as the authentic using different
solvents systems and had the same U.V. absorption. (Tab. 1)

315
Table 1

Absorption maxima {nm} o f Cannabinols

= ~.max (nm)

Cannabinols Hexane Methanol

Cannabidiolic acid 225, 270, 311 224, 266, 304

Cannabidiol 212, 282 213, 283
Tetrahydrocannabinot 212, 234, 283 213, 234, 284
Cannabinol 219, 285 220, 285

Table 2

The cannabinot components in the fractions

collected from the column chromatography

Fraction No. Cannabinols

1- 5 THC and CBC

6- 10 THC, CBC and CBN
11 - 15 THC, CBN and CBD
16 - 20 CBN and CBD
21 - 50 CBD and CBD isomer (CBD')
51 - 70 CBD

Isolation o f the Other Cannabinols

The other cannabinols viz. tetrahydrocannabinol (THC), cannabinol

(CBN), cannabidiol (CBD), cannabichrome (CBC) and another component
(CBD') (giving positive Beam reaction) were isolated from the petroleum
ether-ether eluate, after removing of the cannabidiolic acid, by application
of cotumn and preparative TLC

Column Chromatography

Silica gel, impregnated w i t h dimethylformamide was used (80 g silica gel

'40 mesh' treated with dimethylformamide '40 m l ' , filtered and the
acetone was removed by heating the silica gel at about 35°C till it .smelled
no acetone). One g of the.cannabidiolic acid-free Hashish resin was frac-
tionated on the impregnated silica. Elution was carried out using cyclo-

316
hexane saturated with dimethylformamide and the course of the chromato-
graphic fractionation was followed on plates coated with silica gel G (using
petroleum ether - ether ethyl acetate 90:5:5) and the results obtained
are shown in Table 2. Fractions 1-5, 6-10, 11-15, 16-20 and 2 1 - 5 0
were further fractionated into their cannabinols by preparative TLC.

Preparative TLC

The fractions obtained from the column were fractionated using plates
coated with silica gel G (1 mm thick). The plates, after developing with
petroleum ether - ether - ethyl acetate (90:5:5) were air dried and the
outer strips (about 2 cm width) were sprayed (after covering the central part
with a glass plate ) with tetrazotised o-tolidine. The zones containing the
cannabinols, from the unsprayed central portion were scrapped off and
separately extracted with ether. By applying this technique CBD, CBN,
THC, CBC and CBD' (probably a cannabidiol isomer) (Korte & Sieper, 1965)
were isolated.
The identity of the isolated cannabinols was proved by Rf, using dif-
ferent solvents, and by UV. (Table 1).

Gas-Liquid Chromatography

The purified Hashish resin was investigated by GLC using the following
conditions:
Apparatus: F. & M Model 500, programmed high temperature gas
chromatograph equipped with Model 1609 Flame ionization
detector attachment.
Column dimensions: Copper column 2 meters length and 4 mm diameter.
Solid support: Chromosorb ' 6 0 - 8 0 mesh' (acid washed),
Stationary phase: Apiezon L 20%.
Carrier gas : Nitrogen.
Temperature: 170°C.
Radio-Flowmeter Reading: H2 4.5; air 11.7; N2 2.7.
The results obtained are shown in Fig. 2 and Table 3. The characteristics
,of the Hashish resin chromatogram are compared with the chromatograms
for standard constituents at the same working conditions.

Effect of HCl, Ammonia and Silica Gel on Hashish Constituents,

The effect of these factors on Hashish resin were preliminary studied and
the resulting compounds were detected by two-dimensional TLC. The plates
(20 x 20 cm) were marked by a needle to make two strips each about 5 cm

317
 Table 3

Major components in hashish extract by gasliquid chromatography

Peak No. Retention Identification relative

time (RT) contents
minutes

1 3.6 unknown small

2 4.0 unknown medium
3 5.0 unknown medium
4 5.6 from CBD large
5 7.0 unknown small
6 8.0 from CBD large
7 9.0 unknown medium
8 9.8 unknown large
9 13.6 unknown large
10 14.2 THC medium
11 19.0 CBN medium

wide at the right hand side as well as at the upper part. Hashish resin was
applied on the lower left corner of the plate as well as on each of the strips
to serve as reference for the corresponding development. The plates were
developed with the solvent (petroleum ether - ether - ethyl acetate
90:5:5) in the first direction to a height o f about 14cm, removed from the
chromatographic tanc and air dried. For studying the effect of HC1 or
ammonia vapours, the plate in each case after drying was placed in a jar
saturated with HC1 or ammonia vapours (attained b y placing a beaker con-
taining conc. HC1 or ammonia in the jar) for one hour, then redeveloped in
the second direction with the same solvent system. In order to study the
effect of silica gel adsorbent, the plate after the first development was kept
in dark for 45 hours, then developed in the second direction.
The results obtained (Fig. 3 - 6 ) revealed a remarkable change.

318
LLI
L.J
Z
0
n
l..O 8
ILl
e,"

1
¢r'
0 9
I,--
£.J
ILl
I.--
LIJ
£3

10

11

| I
5 10 15 20

TIME Cminutesl

Fig. ( 2 ) Gas liquid Chromatogram of Purified

Hashish Resin.

319
 .... ' < ] 3 ~ ffl

.']'.

u
.~_

I
)
TNC ) B

CBJI

¢BC
® "g
...... ' 0 ,lz
o X
0
0

CBDA

I.--

u~
~g a:o
._~
v
m

a
0 ;E .c

TH~
D uJ "~
i o

CBN
CBC
CS~T
0
0
~ 0 ..... ...; E
~7-

_
q

E
,~

N
i "-

'~)0
T-
2

320
 ~c
~ o
¢J

(ZZ~
0

._ ~

.=

i
I o

CBO

THC o
CBN
CBC
CBO'

,.,...
,.-,, ,,..,."

CBDA

I-

~ t

O@ ~0ooo
u o

~= -~
ac: '~
Q

W
O.
i
U

2>
o .
C8D ~ o
(D ~ i
THC

CBN
CBc
% ? ,a ~ -.
CBD'
o

•- ~
" ~ ._
CSD~ o
 DISCUSSION

The results obtained from the column chromatographic technique using

different adsorbents and different solvents for elution were unsatisfactory.
Though the column packed with silica gel impregnated with dimethyl-
formamide, using cyclohexane as eluting solvent and collecting fractions
each of 10 ml, gave the best result of these trials, yet only CBD was the only
constituent which obtained sole in some fractions; the other fractions; were
further fractionated by preparative TLC. The latter procedure proved to be
the best.

Of the eleven cannabinols detected in the Hashish resin, six (CBD, CBN,
THC, CBDA, CBD', CBN) were obtained in a pure form and could be
identified. Probably some of the unidentified constituents are isomeric
forms of different cannabinols, particularly tetrahydrocannabinol of which
more isomers were detected and reported to occur in Hashish. The presence
of these isomers was attributed to several factors. A phenolic compound
reported (Korte & Sieper, 1965) to have Rf between CBDA and CBN giving
the same colour reaction as that of CBD and was considered as an isomer of
CBD, probably resulting from the migration of the double bond in the
limonene ring. This compound is reported to exist in considerable amount if
pure CBD is left standing in methanolic solution at room temperature. In
the present work Hashish was macerated with methanol at room tem-
perature for about 24 h and therefore the possibility of the presence of such
isomer is standing. Other factors either genetic or environmental involving
acid, heat, atmospheric moisture, oxygen or light are also responsible for the
formation of tetrahydrocannabinol isomers (Korte & Sieper, 1965).
The use of Apiezon L. as a stationary liquid phase in GLC (not tried
before for cannabinols) has the advantage of using lower temperature and of
being able to obtain better separation. The fact that CBD was the first to
come off the column followed by THC, then CBN, is in agreement with that
in the literature using other columns. The GLC of CBD shows two peaks;
the same observation was obtained by Farmilo et al. (1963) who stated that
these are probably degradation products. The components given by the
standard CBD are represented in the Hashish extract whose chromatogram
shows 11 peaks.

ZUSAMMENFASSUNG

Die Trennung von Haschisch Bestandteilen sowie i_hreIsolierung dutch physikalisch-

chernische Methoden z.B. L6sungsmittel Verteitung, Saulen-und Dtinnschicht-Chroma-
tographie wurde untersucht. Sechs Komponenten: Cannabidiol~ure, Cannabidiol,
Cannabinol, Tetrahydrocannabinol, Cannabichromene und eine positive Beam Kom-

322
ponente (Cannabidiol isomer) wurden getrennt und durch Chromatographie und
UV-Spectrophotometrie nachgewiesen. Die Trennung der Haschisch-Komponenten
gelang auch durch GLC. Der Einfluss einiger Faktoren wie Silica Gel, S//ure und Alkali
wurde studiert.

RI~SUME

La s@aration des constituants du Hashish et l'isolement de certains constituants par

une m6thode preparative a ~t~ effectu~ utilisant plusieurs techniques physico-
chimiques viz. s~paration au solvent, chromatographie sur colmne et sur couche mince.
Six constituants viz. acide cannabidiolic, cannabidiol, cannabinol, t6trahydrocanna-
binol, cannabichromene et un produit positif au r6actif de Beam (isom~re du canna-
bidiol) ont 6t~ isold et identifi~ selon les r~sultats obtenus par chromatographie et par
spectrophotom~trie ultra-viotette. La s~paration des constituants du Hashish a aussi gt~
effectu~ par la chromatographie gas liquide. L'effect de certains facteurs viz. le gel de
silica, l'acidit~ et la basicit~ ont gt~ ~tudi~ rfiv~lant un changement assez notoire.

ACKNOWLEDGEMENT

The authords are deeply indebted to Late Prof. Z. F. Ahmed for his very kind help
and fruitful suggestions. Thanks are to the Central Narcotic Division, Cairo for kindly
supplying Hashish and to Prof. F. Korte and Dr. D. Bieniek (Bonn) for kindly
supplying the authentic references.

REFERENCES

1. Adams, R., Harfenist, M. & L/Swe, S. (1949). New analogues of tetiahydrocanna-

binol XIX. J. Amer. Chem. Soc. 71 : 1624.
2. Beam, W. (1915). Tests for hashish. 4 th Rept. Wellcome Trop. Res. Lab. Chem.
Sect. (Sudan), No. 3, 2 pp.
3, Bergel, F., Morrison, A. L., Rinderknecht, H,, Todd, A. R., Macdonald, A. D. &
Woolfe, G. (1943). Cannabis indica Part XII. Some analogues and a water-soluble
derivative of tetrahydrocannabinol. J. Chem. Soc. 286:
4. Caddy, B., Fish, F. & Wilson, W. D. C. (1967). Gas chromatography of Indian
hemp (Cannabis sativa}. £ Pharm. PharmacoL 19 : 851.
5. Cahn, R. S. (1930). CXXI. Cannabis indica resin. Part I. The constituents of
nitrocannabinolactone (Oxycannabin). J. Chem. Soe. 986.
6. Cahn, R. S. (1931). LXXXVI. Cannabis indiea resin. Part II. J. Chem. Soc. 630.
7. Cahn, R, S. (1932). Cannabis resin. Part III. The consitution of cannabinol. J.
Chem. Soc, 1342.
8. Cahn, R. S. (1933). Cannabis indiea resin. Part IV. The synthesis of some 3:2-
dimethylbenzopyrans and confirmation of cannabinoL 3. Chem, Soc. 1400.
9. Chopra, t. C. & Chopra, R. N. (1957). Bull. Narcot. 9 : 1,4; cited in an article
about Hashish presented by I.C. Chopra to the National Centre for Social and
Criminological Research, Cairo (1970).
10. Claussen, U., Borger, W. & Korte, F. (1966). Zur chemischen Klassifizierung yon
Pflanzen XXVII. (Haschisch, V I ) - Gaschromatographische Analyse der In-
haltstoffe des Hanfes. LiebigsAnn. Chem. 693 : 158.
11. Claussen, U. & Korte, F. (1968). Haschisch, XXVI. Phenolische Inhaltstoffe der

323
 Hanf-Pflanze und ihre Umwandlung zu Haschisch-Inhaltstoffen. Liebigs Ann.
Chem. 713 : 166.
12. Claussen, U., Spulak, F. V. & Korte, F. (1966). Zur chemischen Klassifizierung
von Pflanzen XXXI. Haschisch X. Cannabichromen, ein neuer Haschisch-Inhalts-
Stoff. Tetrahedron. 22 : 1477.
13. Ctaussen, U., Spulak, F. V. & Korte, F. (1967). Haschisch XIV. Tetrahedron. 24 :
1021.
14. Duquenois, P. & Moustafa, H. N. (1938). Identification and assay of Cannabis
indica, or. Egypt. Med. Assoc. 21 : 224.
15. Farmilo, C. G., Davis, T. W. M., Mandenheuvel, F. A. & Lane, R. (1962).
Chemical analysis of marihuana, IV. Biogenesis, paper chromatography, gas
chromatography. Proc. Can. Soc. Forensic ScL 1, paper 2.
16. Fietz-David, H. E. & Blangy, L. (1949). Fundamental process of dye chemistry.
Interscience Publisher Inc., New York.
17. Gaoni, Y. & Mechoulam, R. (1964). Hashish I i t - Isolation, structure and partial
synthesis of an active constituent of hashish. Z Amer. Chem. Soc. 86 : 1646.
18. Gaoni, Y. & Mechoutam, R. (1966). Cannabichromene, a new active principle in
hashish. Chem. Commun. (1) : 20.
19. Grlic, L. (1970). Simple thin-layer chromatography of cannabinoids by means of
silica gel sheets treated with amines. J. Chromatog. 48 : 562.
20. Gibbs, H. D. (1927). Phenol tests, I I I - The indophenol test. J. Biol. Chem. 72 :
649.
21. Hamilton, H. C., Lescohier, A.W. & Perkins, R.A. (1913). The physiological
activity of Cannabis sativa. J. Amer. Pharm. Assoc. 2 : 322.
22. Heaysman, L. T., Walker, E. A. & Lewis, D.T. (1967). The application of gas
chromatography to the examination of the constituents of Cannabis sativa.
Analyst. 92 : 450.
23. Hilvely, R. L., Mosher, W. A. & Hoffman, F. W. (1966). Isolation of trans - A6
-tetrahydrocannabinol from marijuana. J. Amer. Chem. Soc. 88 : 1832.
24. Kane, V. V. & Razdan, R. K, (1968). Constituents of hashish. A novel reaction of
olivetol with citral in the presence of pyridine. Total synthesis of d 1 cannabi-
cycol and d 1 cannabiehromene. J. Amen. Chem. Soc. 90 : 6551.
25. Korte, F., Haag, M. & Claussen, U. (1965). Zur chemischen Klassifizierung von
Pflanzen. XXVIII. Haschisch VII-Tetrahydrocannabinol-carbonsaiire, ein neuer
Haschisch-Inhaltstoff. Angew. Chem. 77 : 862.
26. Korte, F. & Sieper, H. (1964 a). Zur chemischen Ktassifizierung yon Pflanzen.
XXIV. Untersuchung yon Haschisch-lnhaltstoffen dutch Dttnnschichtchromato-
graphic. J. Chromatog. 13 : 90.
27. Korte, F. & Sieper, H. (1964 b). Zur chemischen Klassifiziemng yon Pflanzen.
XXV. Bestimmung yon Haschisch-Ingaltstoffen durch Diinnschhchtchromato-
graphie. J. Chromatog. 14: 178.
28. Korte, F. & Sieper, H. (1965). Recent results of hashish analysis; cited in
'Hashish, its chemistry and pharmacology', Ciba Foundation Study Group No. 21,
Churchill, London (Eis. G. E. W. Wolstenholme and J. Knight) 15.
29. Krejci, Z. (1961). Substances with antibacterial and hashish effect in hemp.
Casopis, L ekaru Ceshy ch. 100 : 1351.
30. Krejci, Z., Horak, M. & Santavy, F. (1958). Constitution of the cannabidiolic
acid and an acid of the m.p. 133 °, isolated from Cannabis sativa. Acta Univ.
Palackianae Olomucensis. 16 : 9.
31. Krejci, Z., Horak, M. & Santavy, F. (1959). Cannabis sativa, an antibiotic medi-

324
 cinal agent III. Isolation and constitution of two acids obtained from C. sativa.
Pharmazie. 14 : 349.
32. Krejci, Z. & Santavy, F. (1955). The isolation of further substances from the
leaves of Indian hemp (Cannabis sativa L. var. indiea}. Acta Univ. Palackianae
Olomucensis. 6 : 59.
33. Mechoulam, R. (1964). Cannabis. Harokeah Hawri. 10 : 96.
34. Mechoulam, R. Ben-Zvi, Z., Yagnitinskey, B. & Shani, A. (1969). New tetra-
hydrocannabinolic acid. Tetrahedron Left. 2339.
35. Mechoulam, R. & Gaoni, Y. (1965). Hashish IV, Isolation and structure of canna-
binolic, cannabidiolic and cannabigerolic acids. Tetrahedron. 21 : 1223.
36. Mechoulam, R. & Gaoni, Y. (1967). Recent advances in the chemistry of Hashish.
Fortsehr. Chem. Org. Naturstoffe. 25 : 175.
37. Morill, F. T. (1938). Marihuana, the new dangerous drug. Washington, Foreign
Policy Assoc. Inc.
38. Obata, Y. & Ishikawa, Y. (1966). Constituents of hemp plant, I I I - Isolation of
Gibbs-positive compound from Japanesehemp.Agr. Biol. Chem. (Tokyo). 30 : 6i9.
39. Okamoto, K. (1967). Compounds of Japanese Cannabis. Kagaku Keisatsu
Keukyush Kokoku. 20 : 109.
40. Schultz, O. E. & Haffner, G. (1958). A sedative principle in German hemp.
(Cannabis sativa). Arch. Pharm. Berl. 291 : 391.
41. Schultz, O. E. & Haffner, G. (1959). A sedative and antibacterial principle in
German hemp. (Cannabis sativa). Z. Naturforsch. 146 : 98.
42. Shoyama, Y., Fujita, T., Yamauchi, T. & Nishioka, I. (1968). Cannabis I I - Canna-
bichromenic acid, a genuine substance of cannabichromene. Chem. Pharm. BulL
(Tokyo). 16 : 1157.
43. Shoyama, Y., Yamauchi, T. & Nishioka, I. (1970). Cannabis V - Cannabigerolic
acid monomethylether and cannabinolic acid. Chem. Pharm. Bull (Tokyo). 18 :
1327.
44. Spulak, F. V., Claussen, U., Fehlhaber, H.W. & Korte, F. (1968). Haschisch
X I X - Tetrahydrocannabitriol-cannabidiolcarbon-s~iureester, ein neuer Haschisch-
tnhaltstoff. Tetrahedron. 24 : 5379.
45. Stone, H. M. (1969). An investigation into forensic chemical problems associated
with Cannabis. United Nations Secretariat Document ST/SoA /SER, S/18.
46. Turk, R. E., Daharir, H. I. & Forney, R. B. (1969 a). Simple chemical method to
identify marihuana. Z Forensic Soft 14 : 349.
47. Turk, R. E. Forney, R. B., King, L. G. & Ramachandram, S. (1969 b). Extraction
and chromatographic isolation, purification and identification of tetrahydrocan-
nabinol and other compounds form marihuana. J. Forensic ScL 14 : 385.
48. Vollner, L , Bieniek, D. & Korte, F. (1969). Haschisch X X - Cannabidiverin, ein
neuer Haschisch-Inhaltstoff. Tetrahedron Lett. 145.
49. Watt, J. M. & Breyer-Brandwijk, H.G. (1962). The Medicinal and poisonous
plants of Southern and Eastern Africa. E. & S. Levingston Ltd. 2nd ed.
50. Wolf, P. O. (1949). Marihuana in Latin America, the threat it constitutes',
Washington. Linacre Press.
51. Yamauchi, T., Shoyama, Y., Aramaki, H., Azuma, T. & Nishioka, I. (1967).
Tetrahydrocannabinolic acid, a genuine substance of tetrahydrocannabinol.
Chem. Pharm. Bull. (Tokyo). 15 : 1075.
52. Yamauchi, T., Shoyama, Y., Matsuo, Y. & Nishioka, I. (1968). Cannabis llI.
Cannabigerol monomethyl ether, a new component of hemp. Chem. Pharm. Bull.
(Tokyo). 16 : 1164.

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