Distribution of Chemical Phenotypes (Chemotypes) in European Agricultural Hemp (Cannabis sativa L.) Cultivars.

In Europe, more than 50 approved cultivars of fiber hemp (Cannabis sativa L.) are in agricultural production. Their content of psychoactive tetrahydrocannabinol (THC) is legally restricted to <0.2% (%w/w in the dry, mature inflorescences). Cannabis strains with much higher THC contents are also grown, illegally or under license for drug production. Differentiation between these two groups relies on biochemical quantification of cannabinoid contents in mature floral material. For nonflowering material or tissue devoid of cannabinoids, the genetic prediction of the chemical phenotype (chemotype) provides a suitable method of distinction. Three discrete chemotypes, depending on the ratio of THC and the noneuphoric cannabidiol (CBD), can be distinguished: a "THC-predominant" type, a "CBD-predominant" type, and an intermediate chemotype. We present a systematic genetic prediction of chemotypes of 62 agricultural hemp cultivars grown in Europe. The survey reveals the presence of up to 35% BT allele-carrying individuals (representing either a THC-predominant or an intermediate chemotype) in some cultivars-which is unexpected considering the legal THC limit of 0.2% THC. The fact that 100% of the seized drug-type seeds in this study revealed at least one BT allele, reflects that plant breeding efforts have resulted in a fixation of the BT allele in recreational Cannabis. To guarantee a sincere forensic application based on a genetic chemotype prediction, we recommend not to classify material of unknown origin if the samples size is below nine genetically independent individuals.

Improving the Performance of High-Precision qNMR Measurements by a Double Integration Procedure in Practical Cases.

Quantitative (1)H NMR (qNMR) is a widely applied technique for compound concentration and purity determinations. The NMR spectrum will display signals from all species in the sample, and this is generally a strength of the method. The key spectral determination is the full and accurate determination of one or more signal areas. Accurate peak integration can be an issue when unrelated peaks resonate in an important integral region. We describe a "hybrid" approach to signal integration that provides an accurate estimation of signal area, removing the component(s) that may arise from unrelated peaks. This is achieved by using the most accurate integration method for the region and removing unwanted contributions. The key to this performing well, and in almost all cases, is the use of areas from deconvolved peaks. We describe this process and show that it can be very successfully applied to cases where the highest precision is required and for more common cases of NMR-based quantitation.