Fatty acid composition of Achene oils from five Moroccan climatic cultivars of Cannabis

Hemp {Cannabis sativa L.) an herbaceous annual plant is of economic importance as a drug, a fibre and oil seed plant. It's considered as one of the oldest of cultivated plants and appears to have originated in northern China (Huí Lin Li, 1974), where it grew in the wild state still around 5200 to 6200 years ago. Actually hemp spread throughout most of the temperature and tropic regions of the world. Both genetic and environmental conditions influenced the biosynthesis process in hemp plant, essentially cannabinoids composition. Warm, dry and windy conditions were believed to induce a higher density of resin glands where the biosynthesis of cannabinoids takes place (Crombie, 1977; Valle et al., 1978; Braut-Boucher, 1980; Mahlberg and Hemphill, 1983). Murari et al., (1983), estimated higher contents of cannabinoids, in the same varieties, when hemp grown in a continental climate than in a maritime climate. One of many compounds secondary metabolites produced by hemp is oil, resulting from the plant lipid synthesis and achenes are the primary storage tissue for accumulating oil. Achenes of Cannabis contain around 30-35% oil, which is brownish-green in colour and smells of the seed (Vaughan, 1970). The oil possesses marked drying properties, and it's used in the manufacture of paint, soaps, varnishes, printing inks and an edible oil. Many countries cultivate hemp as an oil-seed plant, like Manchuria, Chile, Turkey, China, Formosa, Hungary, Czechoslovakia and Francia, the main world cultivator is the ancient USSR. The average annual production of seed being in the region was 250.000 tons (Altschul cited in Vaughan, 1970). In previous studies (Merzouki and Molero Mesa, 1995; Merzouki et al., 1996a and b) about Cannabis chimiotaxonomy (seed morphology and cannabinoids composition), we have concluded that Cannabis cultivated in the Rif belongs to the drug phenotype and the genus was monotype {Cannabis sativa L). The Rif, space of Canna/̂ /s cultivation in Morocco, is characterized by a high ecological variety where bioclimates vary from arid to perhumid (Ben Abid, 1982). The purpose of this study was to determine oil concentrations and differences in the fatty acid composition of the oil in achene from the five «Cannabis climatic varieties» cultivated in the north of Morocco.


INTRODUCTION
Hemp {Cannabis sativa L.) an herbaceous annual plant is of economic importance as a drug, a fibre and oil seed plant.It's considered as one of the oldest of cultivated plants and appears to have originated in northern China (Huí Lin Li, 1974), where it grew in the wild state still around 5200 to 6200 years ago.Actually hemp spread throughout most of the temperature and tropic regions of the world.
Both genetic and environmental conditions influenced the biosynthesis process in hemp plant, essentially cannabinoids composition.Warm, dry and windy conditions were believed to induce a higher density of resin glands where the biosynthesis of cannabinoids takes place (Crombie, 1977;Valle et al., 1978;Braut-Boucher, 1980;Mahlberg and Hemphill, 1983).Murari et al., (1983), estimated higher contents of cannabinoids, in the same varieties, when hemp grown in a continental climate than in a maritime climate.
One of many compounds secondary metabolites produced by hemp is oil, resulting from the plant lipid synthesis and achenes are the primary storage tissue for accumulating oil.Achenes of Cannabis contain around 30-35% oil, which is brownish-green in colour and smells of the seed (Vaughan, 1970).The oil possesses marked drying properties, and it's used in the manufacture of paint, soaps, varnishes, printing inks and an edible oil.Many countries cultivate hemp as an oil-seed plant, like Manchuria, Chile, Turkey, China, Formosa, Hungary, Czechoslovakia and Francia, the main world cultivator is the ancient USSR.The average annual production of seed being in the region was 250.000 tons (Altschul cited in Vaughan, 1970).
In previous studies (Merzouki and Molero Mesa, 1995;Merzouki et al., 1996a and b) about Cannabis chimiotaxonomy (seed morphology and cannabinoids composition), we have concluded that Cannabis cultivated in the Rif belongs to the drug phenotype and the genus was monotype {Cannabis sativa L).
The Rif, space of Canna/^/s cultivation in Morocco, is characterized by a high ecological variety where bioclimates vary from arid to perhumid (Ben Abid, 1982).
The purpose of this study was to determine oil concentrations and differences in the fatty acid composition of the oil in achene from the five «Cannabis climatic varieties» cultivated in the north of Morocco.
Achene samples were milled and the oil was extracted n-hexane in a Soxhiet apparatus.The extract was dried over anhydrous sodium sulphate and the solvent removed under reduced pressure in a rotary film evaporator.Fatty acid composition of solvent-extracted oils from achenes was determined using transesterification of the triacylglycerols with a solution 0,5N KOH in methanol (Sonanini and Weber in Lawi-Berger, 1982) and the methyl esters analysed by gas chromatography uing a glass column (20 m long and 2.25 mm ID) coated with SP-2330.The detector was a flame ionization detector.A fatty acid standard containing the fatty acids myristic (C:14), palmitic (CI 6:0), stearic (CI 8:0), oleic (CI 8:1), linoleic (CI 8:2), linolenic (CI 8:3), arachidic (C20), behenic (C22:0), lignoceric (C24:0).Fatty acid peaks were identified by comparing the fatty acid methyl ester peaks and retention times of standards with sample peaks.Standard solution of margarle acid methyl ester was used for quantification.Seed oil and fatty acid concentrations were determined from one sample per population (Five samples per locality).Means and standard deviation of means are presented to give some ideas of the variability of oil and fatty acids within each variety.Variance analyse (ANOVA) was used to compare oil and fatty acid concentrations between varieties.Correlation analyses were used to examine the relationships among fatty acid composition and ecological and geographical parameters [T° Max.(°C), T" min.(°C), precipitation (mm), altitude (m), latitude and longitude].

Oil concentrations of 5 «climatic
Cannabis varieties» from the Rif are summarized in Table I.The fatty acid composition from Cannabis achenes is summarized in Table III and Fig. 1.The major fatty acids observed were CI 8:1, CI 8:2 and C18:3 in each variety.The lowest one is C24 which varied from 0.31 in Ketama to 0.76 pMol/g oil in Chaouen.Results of variance analysis were summarized in Table IV.Difference analysis of each fatty acid between localities reveal that fatty acid concentrations from Bouhmed achene variety presents a clear difference with the other varieties.The myristic, palmitic, oleic, linoleic, behenic and lignoceric acids from Bouhmed achene variety are a highly significant difference to the others varieties.Achene oil from Bab Taza location are characterized by 3 fatty acids, oleic (C18:1), linoleic (C18:2) and arachidic (C20).These fatty acids are significantly different to thus from others localities.Seller (1986) concluded that environmental factors, especially temperature, during the period of achene development and maturation affect both the concentration and composition of oil in maturing sunflower achenes.Therefore, differences in fatty acid composition found in Cannabis achene oils can be attributed to environmental factors.
Results of Rifian Cannabis varieties show that the achene oils have similar fatty acid composition than the varieties analysed by Lawi-Berger (1982), who has compared fibre and drug varieties.In such study, the author reveal that comparison of fatty acid (c) Consejo Superior de Investigaciones Científicas Licencia Creative Commons 3.0 España (by-nc) http://grasasyaceites.revistas.csic.es

Fatty
Figure 1 Fatty acid concentrations of 5 Cannabis climatic varieties from Morocco

Table IV Difference analysis of means of fatty acid concentrations between all localities Kétama Jebha Bouhmed Chaouen Bab Taza
Cannabis variety.The variability observed in fatty acids of the five Cannabis varieties cultivated in the Rif region was undoubtedly caused by differences in environment at different locations.