Observations on cannabinoid content in Cannabis sativa L. grown in Tuscany, Italy

Sections

ABSTRACT
Introduction
Material and methods
Results

Details

Author: E. BERTOL, F. MARI
Pages: 55 to 60
Creation Date: 1980/01/01

Observations on cannabinoid content in Cannabis sativa L. grown in Tuscany, Italy

E. BERTOL National Research Council; Forensic Medicine Department, University of Florence, Italy
F. MARI
Professor of Toxicological Chemistry, Forensic Medicine Department, University of Florence, Italy

ABSTRACT

The paper summarizes the analytical results on 114 Cannabis sativa L. samples seized in Tuscany. The percentage of Δ 9-tetrahydrocannabinol content in adult plants and the phenotype ratio of young plants were determined to differentiate fibre-type from drug-type cannabis plants. The analysis revealed that 71.9 per cent of the total sample were cannabis plants of a drug type.

Introduction

Use of cannabis in Italy has been increasing in recent years. The number and size of seizures of cannabis have also increased, and in this respect the Tuscan situation, particularly in the Florence area, is similar to the national pattern. Compared with 1975, cannabis exhibits (marijuana, hashish, cannabis oil) increased 56 per cent in 1976, 106 per cent in 1977, 306 per cent in 1978 and 487 per cent in 1979. The Italian law of 1975 (No. 685-22.12) forbids the cultivation of "Canapa Indiana". In the opinion of the authors, this is not an appropriate term [ 1] , [ 2] , though it indicates the psychoactive plant. It is therefore of great importance to make a distinction between plants of cannabis-drug and cannabis-fibre type. This paper presents the results obtained from analyses of seizures of cannabis plants.

Material and methods

The material consisted of both adult and young plants. In adult plants the cannabinoid content was measured as a percentage. In young plants the phenotype ratio (percentage of cannabinol (CBN) + percentage of Δ 9-tetrahydrocannabinol (Δ 9-THC) divided by percentage of cannabidiol (CBD)) was used to differentiate between drug-type and fibre-type cannabis plants. When the phenotype ratio was greater than 1.0 the plant was classified as drug-type; when less than 1.0 it was classified as fibre-type [ 3] .

A total of 114 samples of cannabis seized in Tuscany were selected for analysis. The stems were separated and discarded and the material dried at 60°C for 24 hours. A modified version of the method developed by Patterson and Stevens [ 4] was employed. An amount of 1 g of powdered sample was added to 20 ml of petroleum ether in glass-stoppered tubes. This was then mixed for 5 minutes and maintained for 2 hours at room temperature. Thereafter the insoluble plant material was removed by centrifugation, the solvent was evaporated and the residue redissolved in 1 ml of cyclohexane containing α-cholestane in a concentration of 3 mg/ml as an internal standard. A 1- μl sample of the final solution was injected for gas-liquid chromatographic (GLC) analysis.

Analyses were performed using a Tracor-550 gas chromatograph equipped with a flame-ionization detector and a disc integrator with automatic printer. It was operated isothermally at 220°C (nitrogen flow, 50ml/min; air, 400ml/min; hydrogen, 40 ml/min), with an inlet temperature of 270°C, using a glass column (1/8 in. (3 mm) internal diameter, 2 m long) packed with SE 30 (1%) on Anakrom, 90-100 mesh. Before use, the column was silanized at a low nitrogen flow. Reference solutions were cannabidiol (0.218 mg/ml, 0.436 mg/ml, 0.872 mg/ml, 1.744 mg/ml, 3.488 mg/ml), Δ 9-tetrahydrocannabinol(0.283 mg/ml, 0.566 mg/ml, 1.132 mg/ml, 2.264 mg/ml, 4.528 mg/ml), and cannabinol (0.273 mg/ml, 0.546 mg/ml, 1.092 mg/ml, 2.184 mg/ml, 4.368 mg/ml). Each reference solution contained α-cholestane at a concentration of 3 mg/ml. The results of the analyses are summarized in the table.

Cannabinoid content in Cannabis sativa L.

Sample No.

Age of plant

CBD (%)

Δ9-THC (%)

CBN (%)

Phenotype ratio

Type

Origin

1
Y
0.576 0.038 0.118 0.27
F
 
2
Y
0.206 0.515 0.412 4.5
D
 
3
Y
0.195 0.493 0.401 4.58
D
 
4
Y
0.213 0.564 0.432 4.67
D
 
5
Y
0.110 0.667 0.544 11.00
D
 
6
Y
0.027 0.104 0.2 11.25
D
 
7
Y
0.035 0.113 0.234 9.91
D
 
8
Y
0.09 0.039 0.266 3.38
D
 
9
Y
0.05 0.032 0.194 4.52
D
 
10
Y
0.130 0.571 0.553 8.64
D
A
11
Y
0.032 0.127 0.19 9.9
D
R
12
Y
0.209 0.253 0.073 1.55
D
E
13
Y
0.051 0.087 0.233 6.27
D
A
14
Y
0.150 0.490 0.849 8.92
D
I^
15
Y
0.061 0.095 0.251 5.67
D
 
16
Y
0.239 0.286 0.065 1.46
D
 
17
A
0.214 1.11
t
-
D
 
18
A
0.099 1.42 0.09
-
D
 
19
A
0.271 1.477 0.206
-
D
 
20
A
0.321 1.98 0.514
-
D
 
21
A
0.032 0.976
t
-
D
 
22
A
0.142 0.818 0.232
-
D
 
23
A
0.02 0.993 0.051
-
D
 
24
A
0.028 0.920 0.023
-
D

Sample No.

Age of plant

CBD (%)

Δ9-THC (%)

CBN (%)

Phenotype ratio

Type

Origin

25
A
0.021 0.874
t
-
D
 
26
A
0.10 1.907
t
-
D
 
27
Y
0.493 0.032 0.093 0.25
F
 
28
Y
1.09 0.161 0.065 0.20
F
 
29
Y
0.479 0.029
t
0.06
F
 
30
Y
0.583 0.041 0.122 0.27
F
 
31
Y
0.460 0.055 0.026
0. 17
F
 
32
Y
1.13 0.185 0.072 0.22
F
A
33
Y
0.691 0.383 0.028 0.59
F
R
34
Y
0.542 0.078 0.030 0.19
F
E
35
Y
0.058 0.285 0.114 6.87
D
A
36
A
0.048 0.035
t
-
F
II^
37
A
0.653 0.15 0.018
-
F
 
38
A
0.089 0.07
t
-
F
 
39
A
0.975 0.25
t
-
F
 
40
A
0.035 0.683 0.03
-
D
 
41
A
0.114 0.772 0.323
-
D
 
42
Y
0.71 0.31
t
0.43
F
 
43
Y
0.03 0.697
t
23.2
D
A
44
Y
0.195 0.522 0.106 3.22
D
R
45
A
0.350 2.452
t
-
D
E
46
A
0.048 1.02
t
-
D
A
47
A
0.097 1.486 0.025
-
D
III^
48
Y
2.5
t
t
0.00
F
 
49
Y
0.413 0.022 0.03 0.12
F
 
50
Y
1.03 0.140 0.035 0.16
F
 
51
Y
0.713 0.063 0.031 0.13
F
 
52
Y
0.853 0.478
t
0.56
F
 
53
Y
0.638 0.069 0.042 0.17
F
A
54
Y
1.03 0.047 0.02 0.06
F
R
55
Y
1.09 0.13 0.25 0.34
F
E
56
Y
0.632 0.042 0.134 0.27
F
A
57
Y
0.405 0.035 0.042 0.19
F
IV^
58
A
0.135 0.054 0.03
-
F
 
59
A
0.065 0.03 0.02
-
F
 
60
A
0.175
t
t
-
F
 
61
A
0.093 0.065
t
-
F
 
62
A
0.152 0.723 0.048
-
D
 
63
Y
0.023 0.638
t
27.7
D
 
64
Y
0.20 0.658
t
3.29
D
 
65
Y
0.142 0.501 0.705 8.49
D
 
66
Y
0.11 0.75
t
6.81
D
 
67
Y
0.03 0.54 0.06 20.0
D
 
68
Y
0.108 0.654 0.532 10.9
D
 
69
Y
0.083 0.045 0.231 3.32
D
A
70
Y
0.06 0.028 0.213 4.01
D
R
71
Y
0.191 0.205 0.06 1.38
D
E
72
Y
0.07 0.123 0.253 5.37
D
A
73
Y
0.075 0.083 0.151 3.12
D
V^
74
Y
0.03 0.065 0.251 10.5
D
 
75
A
0.89 1.29 1.009
-
D
 
76
A
0.02 1.12 0.206
-
D
 
77
A
0.543 1.19 0.14
-
D
 
78
A
0.227 1.4 0.082
-
D
 
79
A
0.09 1.31 0.04
-
D
 

Cannabinoid content in Cannabis sativa L. (continued)

Sample No.

Age of plant

CBD (%)

Δ9-THC (%)

CBN (%)

Phenotype ratio

Type

Origin

80
A
0.091 1.00
t
-
D
 
81
A
0.035 0.927 0.02
-
D
 
82
A
0.227 1.42 0.07
-
D
 
83
A
0.03
t
t
-
F
 
84
Y
0.250 0.570 0.030 5.6
D
 
85
Y
0.04 0.351 0.125 11.9
D
 
86
Y
0.02 0.421
t
21.05
D
 
87
Y
0.242 0.453 0.09 2.24
D
A
88
Y
0.035 0.729
t
20.8
D
R
89
Y
0.238 0.856
t
3.59
D
E
90
Y
0.027 0.642 0.053 25.7
D
A
91
Y
0.130 0.783 0.851 12.56
D
VI^
92
Y
0.042 0.585 0.19 18.45
D
 
93
A
0.047 1.25 0.03
-
D
 
94
A
0.09 0.06 0.02
-
F
 
95
Y
0.025 0.482 0.055 21.48
D
 
96
Y
0.132 0.427 0.041 3.54
D
 
97
Y
0.061 0.593 0.019 10.03
D
 
98
Y
0.047 0.635 0.076 15.12
D
 
99
Y
0.042 0.31 0.35 15.71
D
 
100
Y
0.065 0.328 0.115 6.81
D
 
101
Y
0.071 0.095 0.163 3.63
D
 
102
Y
0.263 0.531 0.08 2.32
D
A
103
Y
0.096 0.054 0.210 2.75
D
R
104
Y
0.029 0.756
t
26.06
D
E
105
A
0.119 1.45 0.14
-
D
A
106
A
0.027 1.85 0.105
-
D
VII^
107
A
0.167 2.2
t
-
D
 
108
A
0.213 2.47 0.327
-
D
 
109
A
0.091 1.92
t
-
D
 
110
A
0.085 0.93
t
-
D
 
111
A
t
1.03
t
-
D
 
112
A
0.09 0.987 0.063
-
D
 
113
A
0.049 0.035
t
-
F
 
114
A
0.035
t
0.029
-
F
 

Note: Y = young; A = adult; F = fibre-type; D = drug-type; t = trace.

Results

Analysis showed that 71.9 per cent of the cannabis samples were drug-type plants and only 28.1 per cent were fibre-type plants.

On the basis of the phenotype ratio it was found that 49 samples of young plants, or 71.0 per cent, were drug-type. Among adult plants, the determination of Δ 9-THC content showed that 33 samples, or 73.3 per cent, were drug-type plants.

On the basis of the above analysis of cannabis samples, seven geographical areas in the region of Tuscany were identified as presented in the figure. In five of these areas cannabis samples were predominantly drug-type, whereas in two they were mainly fibre-type. In most cases the seeds were of Italian origin.

Map of Tuscany showing the areas of seized Cannabis plants

Full size image: 40 kB, Map of Tuscany showing the areas of seized Cannabis plants

References

001

G. F. Phillips, "The legal description of cannabis and related substances", Medicine, Science and the Law, vol. 13, 1973, pp. 139-142.0.

002

E. Small, "American law and the species problem in cannabis: science and semantics", Bulletin on Narcotics (United Nations publication), vol. 27, No. 3 (1975), pp. 1-2

003

P. S. Fetterman and others, "Mississippi-grown Cannabis sativa L.: preliminary observation on chemical definition of phenotype and variations in tetrahydrocannabinol content versus age, sex and plant part", Journal of Pharmaceutical Sciences, vol. 60, 1971,. pp. 1246-1249

004

D. A. Patterson and H. M. Stevens, "Identification of cannabis", Journal of Pharmacy and Pharmacology, vol. 22, 1970, pp. 391-392.