Bulletin on Narcotics - Oct-Dec 1980Die folgende Studie erschien im Jahre 1980 in der Zeitschrift «Bulletin on Narcotics». Sie analysierte den Wirkstoffgehalt von Cannabis aus Beschlagnahmungen in Großbritannien zwischen 1976 und 1978.
Variation in the THC content in illicitly imported Cannabis products
Author: P. B. BAKER, K. R. BAGON, T. A. GOUGH
Laboratory of the Government Chemist, London, United Kingdom of Great Britain and Northern Ireland K. R. BAGON
Laboratory of the Government Chemist, London, United Kingdom of Great Britain and Northern Ireland T. A. GOUGH
Laboratory of the Government Chemist, London, United Kingdom of Great Britain and Northern Ireland
The tetrahydrocannabinol (THC) content of 304 seizures of illicit Cannabis products was determined using gas chromatography. This procedure results in the conversion of THC acid to THC itself and thus gives the total THC content of the material, which reflects its potency as experienced by the smoker. The study was carried out over three years, thus enabling comparisons to be made between THC levels from a given country of origin over this time span in addition to any variations between the countries. In the early part of the study, herbal cannabis from South East Asia was consistently the best quality and that from the Caribbean the poorest quality. In the third year of the study, the material from South East Asia was in general no richer in THC than material from other areas of the world. Cannabis resins normally had higher THC contents than most herbal material, but the highest levels were found in "hash oil" from the Middle East and the Indian subcontinent.
The price of Cannabis products on the illicit market is governed inter alia by their quality as perceived by the user. It has been established for some years that the major psychoactive constituent in Cannabis is Δ 9-tetrahydrocannabinol (THC) and the level of this compound in Cannabis products can therefore be used as a guide to quality.
* Cannabis = Cannabis sativa L.; cannabis = marijuana; cannabis resin = hashish; cannabis oil = liquid cannabis or "hash oil".
The major cannabinoids (THC, cannabinol and cannabidiol) can be separated by thin-layer chromatography. Quantitative information can be obtained by making absorption measurements after reacting the separated constituents with azo dyes. Using this method Nielsen  examined cannabis grown in Denmark and found that the THC content of the dried mature plant varied between 0.04 and 1.7 per cent. The majority of reports describe the use of gas chromatography to separate the cannabinoids, and quantitative information is based on measurements using the flame-ionization detector. Quantitation has been carried out using either internal or external standards with coefficients of variation ranging from 1 to 3 per cent, which is adequate for the present work  - . Reviews of gas chromatographic procedures have been published  ,  in which various stationary phases and operating conditions have been compared. Satisfactory separation of cannabinoids, after extraction, may be obtained using a variety of silicone gum phases. It has, however, been noted that loss of THC by absorption on stationary phases can occur at low THC levels  .
A method has also been described in which the direct injection of solid cannabis is made  . However, there is ample evidence to show that the cannabinoid content varies within a plant  . This method has therefore no advantage over extraction procedures as in order to obtain representative and homogeneous samples a large amount must be prepared from which an aliquot is taken. Ohlsson et al.  have shown that there is no significant difference between the THC levels (and between other cannabinoid levels) in male and female plants grown under the same conditions. We have, however, noticed that the majority of herbal sample seizures submitted to us do not contain a significant proportion of male plants.
The cannabinoid acids are thermally labile and the use of gas chromatography results in their decarboxylation to the corresponding cannabinoids. Quantitative measurement of the THC content by gas chromatography therefore gives the total amount of THC and its acid (THCA), the latter being quantitatively converted to THC on injection  ,  . For the present study this is an advantage because the acids also decarboxylate during the process of smoking. It is possible to determine the THC and THCA content separately by gas chromatography using derivatization  ,  or selective extraction  , although recent developments enable this to be done more readily using high-performance liquid chromatography  ,  ,  . There have been reports of the conversion of cannabidiol (CBD) to THC, or a compound of the same gas chromatographic retention time, and this could give rise to erroneous THC levels using gas chromatography  . Under the conditions used in the present work this conversion was not observed. Changes in the THC content during the growth of the plant have been studied and seasonal variations in THC content have been noted  ,  . The THC content of harvested material decreases on storage and there have been several studies in which plant material from a single plant has been stored under different conditions to determine the effects of temperature, light and air on changes in cannabinoid levels  ,  . Even after solvent extraction, cannabinoid concentrations will still change and analysis should be carried out as soon as possible after preparation of the extract  ,  .
There are several reports in the literature on the THC content of cannabis and these are summarized in table 1. In the present context the origin refers to the country in which the plant was grown and not the country of origin of the seeds. It should be noted that, although all the THC levels quoted are expressed as a percentage of dry weight of cannabis, it is not clear in all cases whether any selection of plant material had taken place. In all but one study, values were obtained by gas chromatography and therefore include THCA. As far as can be ascertained, samples were extracted and analysed within a short space of time so that the effects of sample or extract storage on the THC levels was minimal  . Most of the data in table 1 are derived from cannabis legitimately grown and processed as part of scientific studies on cannabinoids. In contrast, the present work is concerned with the THC content of illicit supplies of Cannabis products and all samples have been taken from seizures by law enforcement agencies in the United Kingdom. Jenkins and Patterson  have extensively studied illicit Cannabis products, but the data are expressed in terms of the relative proportions of THC, CBD and cannabinol (CBN) rather than their concentrations in the plant. The valuable study by Marshman et al.  is restricted to Jamaican cannabis. The origins of the samples analysed by Eskes et al.  , all of which were seizures, are not quoted.
Summary of published data on THC content of mature cannabis
We now report the findings of a study of the THC content of cannabis, resin and oil as received in the United Kingdom from a number of different countries over three one-year periods. The most probable country of origin was assigned by taking into account information from the carrier, visual appearance  , and the thin-layer and high-performance liquid chromatographic characteristics [18, 28]. For completeness, the data for 1975 for cannabis and resin which have been published in the Report of the Government Chemist 1976 have been included.
Only samples which to the best of our knowledge were fresh (i.e. less than three months old) on arrival in the United Kingdom were included in the study. This represents approximately 15 per cent of all illicit Cannabis samples submitted to the laboratory over the same period of time. Most samples were analysed within one month of seizure, during which time they were stored in sealed plastic bags at room temperature. Each seizure was analysed as received, except that in the case of herbal material intact stems were first removed. Resin (5 g) or herbal material (2.5 g) was subjected to Soxhlet extraction using 100 ml of ethanol (PBS grade) and the optimum extraction time was found to lie between 60 and 90 minutes. Excessive time resulted in loss of THC by thermal degradation and too short a time in incomplete extraction. It was necessary to grind the resin prior to extraction in order to recover all the THC. For example, two samples of the same resin, one untreated and the other ground, gave THC values of 2.4 and 4.6 per cent, respectively, when extracted for the same time. Examination of the final extract was made on all samples to ensure that extraction was complete, and in no case was any THC found.
Reproducibility of extraction was measured using nine samples of herbal material from a single seizure and results were sensibly constant. It is known from other studies  that in resin, at least from Pakistan and Lebanon, the cannabinoids are uniformly distributed within a block.
Liquid cannabis ("hash oil"), which varied in viscosity from a freely flowing liquid to almost solid, was warmed and diluted with ethanol to give a 1 mg/ml solution.
Each extract, which was analysed on the same day that it was prepared, was made up to 200 ml in ethanol, a 1-ml aliquot was diluted to 10 ml and 5μl of this solution analysed by gas chromatography. A Pye-Unicam model 104 instrument was fitted with a 1.5 m x 4 mm internal diameter glass column containing 3 per cent OV17 stationary phase on 100-120 mesh Gaschrom Q support. The carrier-gas flow rate was 30 ml/min and the column temperature was 250°C. Detection was by flame ionization and the detector was calibrated for THC response using a primary standard of 0.01 per cent THC in ethanol.
The THC content of plant material, resin and hash oil seizures are given in tables 2-4. For some countries there were very few samples, but it is apparent that the range of THC content varies widely even from a given country of origin. This is to be expected, since even plants grown under carefully controlled conditions show wide variations in THC content (see table 1). However, the quality of cannabis from some areas is consistently better than others. During 1975 and 1976, Ghana yielded cannabis low in THC compared with other African countries, but the samples seen in 1978 were of better quality. Jamaican cannabis was consistently poor in 1975 and 1976 and no material high in THC, such as obtained by other workers from dealers in Jamaica  , was found in the United Kingdom. By contrast, there was a noticeable improvement in the Jamaican material in the third year of the study. Mean values for all samples in the present study were, however, little different from published mean values  ,  .
Total THC content of seized cannabis
Total THC content of seized cannabis resin
Total THC content of seized "hash oil"
In the same two years, by far the highest quality cannabis originated in South East Asia (exclusively in the form of "Thai sticks") and this was reflected in its street price, at least in the United Kingdom, over the same period. However, the 1978 seizures which originated in Thailand, while still prepared in the form of sticks, showed a dramatic decrease in THC level compared with previous years. A careful study of the physical appearance of seizures of Thai origin for the three years revealed an increasing seed content in the cannabis. It is not possible to examine the data for any correlation of THC content with season because our procedure for estimating age will not discriminate between differences of less than three months and because there were, in most cases, insufficient samples upon which to base any such conclusion.
The cannabis resins generally had a higher THC content than most of the herbal material with the exception of the best Indian, Rhodesian and Thai material. For any given country there was a wide range of THC values. Not surprisingly, the highest levels of THC were encountered in the hash oil seizures, with values as high as 40 per cent. A comparison of the mean amounts of THC in cannabis resin and hash oil from the same countries (tables 3 and 4) shows that for India, Lebanon and Pakistan, the oil contains approximately three times the amount present in the resin. By contrast, Moroccan oil, which is the least viscous encountered, has no more THC than resin from the same country.
E. Nielsen, "Thin-layer chromatographic analysis of Cannabis from Danish and other sources", Dansk Tidsskr Farm , vol. 44, No. 10 (1970), pp. 359-364.002
P. Lerner, "The precise determination of tetrahydrocannabinol in marihuana and hashish", Bulletin on Narcotics (United Nations publication), vol. 21, No. 3 (1969), pp. 39-42.003
R. Phillips and others, "Seasonal variations in cannabinolic content of Indian marihuana", Journal of Forensic Sciences , vol. 15, No. 2 (1970), pp. 191-200.004
N-A. Ek and others, "Cannabinoid content of fifty seized hashish samples", Journal of Forensic Sciences , 1972, pp. 456-459.005
E. P. Chiesa, R. V. D. Rondina and J. D. Coussio, "Chemical composition and potential activity of Argentina marihuana", Journal of Pharmacy and Pharmacology , vol. 25, 1973, pp. 953-956.006
J. W. Fairbairn and J. A. Liebmann, "The extraction and estimation of the cannabinoids in Cannabis sativa L. and its products", Journal of Pharmacy and Pharmacology , vol. 25, 1973, pp. 150-155.007
F. Fish, "Gas chromatography of Cannabis constituents", Chromatographia, vol. 7, No. 2 (1974), pp. 311-315.008
J. M. Parker and B. L. Stembal, "Review of gas-liquid chromatography of marihuana", Journal of Association of Official Analytical Chemists , vol. 57, No. 4 (1974), pp. 888-892.009
N. E. Hoffman and R. K-H. Yang, "Gas chromatography of Δ 1-tetrahydro-cannabinol", Analytical Letters , vol. 5, No. 1 (1972), pp. 7-11.000
Variation in the THC content 53010
K.E. Rasmussen, S. Rasmussen and A.B. Svendsen, "Gas-liquid chromatography of cannabinoids in micro quantities of Cannabis by solid injection", Journal of Chromatography , vol. 69, 1972, pp. 381-384.011
J. W. Fairbairn, J. A. Liebmann and M.G. Rowan, "The stability of Cannabis and its preparations on storage", Journal of Pharmacy and Pharmacology , vol. 28, 1976, pp. 1-7.012
A. Ohlsson and others, "Cannabinoid constituents of male and female Cannabis sativa ", Bulletin on Narcotics (United Nations publication), vol. 23, No. 1 (1971), pp. 29-32.013
P. Fetterman and others, "Simple gas-liquid chromatography procedure for determination of cannabinoidic acids in Cannabis sativa ", Experimentia, vol. 27, 1971, pp. 988-990.014
C. E. Turner and others, "Constituents of Cannabis sativa L. VII: Use of silyl derivatives in routine analysis", Journal of Pharmaceutical Sciences , vol. 63, No. 12 (1974), pp. 1872-1876.015
R. N. Smith, "High pressure liquid chromatography of Cannabis: identification of separated constituents", Journal of Chromatography , vol. 115, 1975, pp. 101-106.016
A. N. Masoud and N.J. Doorenbos, "Mississippi-grown Cannabis sativa L. III: Cannabinoid and cannabinoid acid content", Journal of Pharmaceutical Sciences , vol. 62, No. 2 (1973), pp. 313-315.017
P. B. Baker and R. Fowler, "Analytical aspects of the chemistry of Cannabis", Proceedings of the Analytical Division of the Chemical Society, 1978, pp. 347-349.018
P. B. Baker and others, "Determination of the distribution of cannabinoids in cannabis resin using high performance liquid chromatography", Journal of Analytical Toxicology , vol. 4, 1980, in press.019
G. W. Kinzer and others, "The fate of the cannabinoid components of marihuana during smoking", Bulletin on Narcotics (United Nations publication), vol. 26, No. 3 (1974), pp. 41-54.020
J. A. Marshman, R. E. Popham and C. D. Yawney, "A note on the carmabinoid content of Jamaican ganja", Bulletin on Narcotics (United Nations publication), vol. 28, No. 4 (1976), pp. 63-68.021
C. E. Turner and others, "Constituents of Cannabis sativa L. IV: Stability of cannabinoids in stored plant material", Journal of Pharmaceutical Sciences , vol. 62, No. 10 (1973), pp. 1601-1605.022
R. N. Smith and C. G. Vaughan, "The decomposition of acidic and neutral cannabinoids in organic solvents", Journal of Pharmacy and Pharmacology , vol. 29, 1977, pp. 286-290.023
J.M. Parker and others, "Decomposition of cannabidiol in chloroform solution", Journal of Pharmaceutical Sciences , vol. 63, No. 6 (1974), pp. 970-971.024
K. Narayanaswami and others, "Stability of Cannabis sativa L., samples and their extracts, on prolonged storage in Delhi", Bulletin on Narcotics (United Nations publication), vol. 30, No. 4 (1978), pp. 57-69.025
R.W. Jenkins and D.A. Patterson, "The relationship between chemical composition and geographical origin of Cannabis", Forensic Science , vol. 2, 1973, pp. 59-66.026
D. Eskes, A. M. A. Verwey and A. H. Witta, "Thin-layer and gas chromatographic analysis of hashish samples containing opium", Bulletin on Narcotics (United Nations publication), vol. 25, No. 1 (1973), pp. 41-47.027
M. J. de Faubert Maunder, "The forensic significance of the age and origin of Cannabis", Medicine, Science and the Law, vol. 16, No. 2 (1976), pp. 78-90.028
R. Fowler, R. A. Gilhooley and P. B. Baker, "Thin-layer chromatography of cannabinoids", Journal of Chromatography, vol. 171, 1979, pp. 509-511.029
Report of the Government Chemist 1976 (London, H.M. Stationery Office, 1977), p. 65.030
V. Rubin and L. Comitas, Ganja in Jamaica: studies in the social sciences (The Hague, Mouton, 1975), vol. 26.031
I. G. Karniol and E. A. Carlini, "The content of (-)Δ 9-trans-tetrahydro-cannabinol (Δ 9-THC) does not explain all biological activity of some Brazilian marihuana samples", Journal of Pharmacy and Pharmacology, vol. 24, 1972, pp. 833-835.032
J. W. Fairbairn and J. A. Liebmann, "The cannabinoid content of Cannabis sativa L. grown in England", Journal of Pharmacy and Pharmacology, vol. 26, 1974, pp. 413-419.033
J. H. Holley, K. W. Hadley and C. E. Turner, "Constituents of Cannabis sativa L. XI: Cannabidiol and cannabichrome in samples of known geographical origin", Journal of Pharmaceutical Sciences, vol. 64, No. 5 (1975), pp. 892-894.034
Commission of Inquiry into the Non-Medical Use of Drugs, "Cannabis", Interim Report, Information Canada, 1972.