9 Drugs Containing Cannabinoids and Kavapyrones

9.1 Cannabis Herba, Cannabis sativa var. indica L., Cannabaceae

The cannabinoids are benzopyran derivatives. Only A9,lO-tetrahydro-cannabinol(TI-IC)

shows hallucinogenic activity. The type and quantity of the constituents depend on the
geographical origin of the drug, climatic conditions of growth, time of harvesting and
storage conditions.
Marihuana: the flowering or seed-carrying, dried branch tips of the female plant.
Hashish: the resin exuded from the leaves and flower stalks of the femaIe plant.

9.1.1 Preparation of Drug Extracts

Powdered drug (I g) is extracted by shaking at room temperature for 10 min with 10 ml

methanol. The filtrate is evaporated and the residue dissolved in 1 ml toluene; depending
on rhe cannabinoid concentration, 5-50 pl is used for TLC.

9.1.2 Thin-Layer Chromatography

10 mg thymol is dissolved in 10 rnl toluene; 5 p1 is used for TLC.

1 mg synthetic tetrahydrocannabinol (THC) is dissolved in 5 ml CHCI,; 3 p.1 is used for Reference
TLC. solutions

Silica gel 60 F,,,-precoated TLC plates (Merck, Darmstadt) Adsorbent

n-hexane-diethyl ether (80:20) or Chromatography

n-hexane-dioxane (90: 10) solvent

9.1.3 Detection

UV-254 nm Prominent quenching of cannabinoids.

Fast blue salt reagent Cannabinoids appear orange-red or carmine (vis);
(FBS No.15) standard thpmol gives an orange colour.
 9.1.4 Formulae

COOH

Cannabidiol acid (CBDS) Cannabidiol (CBD)

Cannabinol (CBN) A9,10-Tetrahydrocannabinol(THC)

9.2 Kava-Kava, Piperis methystici rhizoma,

Piper methysticum G. FORST., Piperaceae (MD, DAC 86)

Depending on its geographical origin, the drug contains 5%-9% kavapyrones. These are
derivatives of 6-styryl-4-rnethoxy-a-pyrones with anticonvulsive, muscle-relaxing and
generally sedative effects.

9.2.1 Preparation of Drug Extracts for TLC

Powdered drug (0.6 g) is extracted with 10 ml dichloromethane for 10 min under reflux
and 0.5 g of a commercial extract is dissolved in 5 ml methanol; 10 yl of each filtrate is
used for TLC investigation.

9.2.2 Thin-Layer Chromatography

Reference 1 mg kawain is dissolved in 1rnl MeOH; 10 p1 is used for TLC.

Adsorbent Aluminium oxide 60 P,,, (Merck, Darmstadt)

Chromatography n-hexane-ethyl acetate (70:30) (2 X 15 cm)

solvent
 9 Drugs Containing Car~rlabinoidsand Kavapyrones 259

9.2.3 Detection

UV-254 nm Prominent quenching of all kawapyrones.

Spray reagent Anisaldehyde sulphuric acid reagent (AS No.3)
(see Appendix A ) Red to violet-red zones (vis.).

9.2.4 Formulae

Piperis methysrici rhizoma (Kava-Icava)

OCH,

@*'.
R,
R2

Desniethoxy-
yangonin (0,6-1%) R, = R, = H IZawain (t,8-2.1%) R, = R, = H
Yanganin (1-1,7%) R, = OCH,; R, = H MetI~ysticin (1,2-2%) R,, R, = -OCM,O-

Dihydrokawain (0,6-1%) R, = R, = H
Dihydrometl~ysticin (0,5-0,896) R,, R, = -OCH,O-
 9.3 Chromatograms

Cannabis herba, Hashish

Drug sample 1 Hashish (Turkish, 1980) 3 Hashish cigarette
2 Hashish (Iranian, 1980) 4-6 Cannabis herba (drug collection)
Reference T thymol
THC tetrahydrocannabinol (synthetic)
Solvent system Fig. 1 n-hexane-diethyl erher (80:20)
Detection Fast blue salt reagent (FBS No. 15) followed by 0.1 M NaOH+ vis

Fig. I Treatment with FBS-NaOH reagent shows intense red-violet to red-orange zones (vis.).
Hashish samples 1 and 3 show two prominent red zones in the R,range 0.45-0.55 due to
cannabinol (CBN) and cannabidiol (CBD).
Between CBS and CBN, sample 2 has the additional red-violet zone of tetrahydrocanna-
- -
binol (THC) at R, 0.5. The three to four red zones from the start up to R,. 0.15 are due
to cannabidiol acid and other polar cannabinoids.
Cannabis herba samples 4-6 contain cannabinoids in low concentrations only.

Kava-Kava rhixoma, Piper methysticum

Drug sample 1 Kava-kava rhizoma
2 Kava-kava extracturn (trade sample)
Reference TI kawain
Adsorbent Fig. 2 aluminium oxide 60 F,,, plates
Solvent system n-hexane-ethyl acetate (70:30)(+ 2 X 15 cm)
Detection A without chemical treatment + UV-254, nm
B Anisaldehyde-sulphuric acid reagent (AS No. 3) + vis

Fig. 2A The Kava-extracts (1,2) are characterized in UV-254 nm by five lacrones in the R, range
0.4-0.8:
methysticin
dihydromethysticin
kawain (TI)
dihydrokawain
desmethoxyltawain
B The quenching zones in Fig. 2 A appear red to blue-violet after treatment with-the AS
-
reagent. Kawain (TI) is seen as a red-violet zone at R, 0.55, followed by weaker violet
-
- -
zones of dihydrokawain (R, 0.65) and desmethoxy1cawain ( R, 0.75). In the R, range
below kawain, the violet zone of dihydromethysticin (R, 0.45) and methysticin (R, -
0.4) are found. A very weak yellow zone of yangonine can overlap the kawain zone at R,
- 0.6.