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Analysis of cannabinoid profiles and THCAS sequences in seized cannabis recovered from growing farm

The aim of this study was to assess the genetic and chemical profiles of the seized cannabis obtained from the growing farm. According to the farmer, certified variety Santhica 27 was sown. However, police officers recovered 118 plants under the suspicion of drug production. Quantitative cannabinoid content was determined using GC-FID on Agilent 7890A GC System. Extraction of DNA from plant material was performed using DNAeasy Plant Mini kit (Qiagen) following the manufacturer’s protocol. DNA quantity was determined by Qubit™ dsDNA HS Assay Kit (Thermo Fisher Scientific). A portion of the tetrahydrocannabinolic acid synthase (THCAS) gene was amplified using Qiagen Multiplex PCR kit. Sequencing was performed according to the laboratory standard operating procedures using Big Dye Terminator v.3.1 Cycle Sequencing Kit on AB3500xl GA (both from Life Technologies). Data analysis was accomplished in Sequencher v.5.4.6 (Gene Codes). Sequences were aligned against the THCAS coding sequence of the drug-type Skunk deposited in NCBI (KJ469378). Based on qualitative chemotype, 61% of total samples were cannabigerol (CBG) predominant, and 39% were THC dominant. Alignment of the obtained sequences correlated with their cannabinoid content, clustering in two distinct groups. Eleven identical sequences differed from variants in hemp Finola and probably Santhica 27 that showed undetectable THCAS. Moreover, several variants were revealed including 1064A that had been previously reported to be responsible for THCAS inactivation and cannabigerolic acid (CBGA) precursor accumulation. This finding was in correlation with the average CBG content of 2%. The remaining three sequences were identical to those obtained in drug-type. The average proportion of THC in these plants was 2.5%. According to chemical and genetic results, we conclude that on the industrial hemp farm was growing THC and CBG predominant cannabis varieties under the guise of certified variety Santhica 27.

Cannabis sativa L ; chemotype ; THCA synthase ; growing farm

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