Journal:Chemical profiling and characterization of different cultivars of Cannabis sativa L. inflorescences by SPME-GC-MS and UPLC-MS

Full article title	Chemical profiling and characterization of different cultivars of Cannabis sativa L. inflorescences by SPME-GC-MS and UPLC-MS
Journal	Separations
Author(s)	Cicaloni, Vittoria; Salvini, Laura; Vitalini, Sara; Garzoli, Stefania
Author affiliation(s)	Toscana Life Sciences Foundation, Università degli Studi di Milano, Sapienza University
Primary contact	Email: v dot cicaloni at toscanalifesciences dot org
Editors	Miguel Ángel Rodríguez-Delgado
Year published	2022
Volume and issue	9(4)
Article #	90
DOI	10.3390/separations9040090
ISSN	2297-8739
Distribution license	Creative Commons Attribution 4.0 International
Website	https://www.mdpi.com/2297-8739/9/4/90/htm
Download	https://www.mdpi.com/2297-8739/9/4/90/pdf?version=1648894245 (PDF)

Abstract

The chemical profile of Cannabis sativa L. female inflorescences is rather complex, being characterized by a large number of molecules belonging to different chemical classes. Considering the numerous applications of cannabis in various fields—including the medical and pharmaceutical sectors, which have seen an increasing use for the Cannabis genus in recent years—a precise characterization of the matrices is essential. In this regard, the application of adequate and suitable sampling and analysis techniques becomes important in order to provide an identification of the metabolites characterizing the profile of the sample under examination.

The goal of this work is to provide additional information on the chemical composition of the inflorescences of five different C. sativa cultivars grown in Emilia Romagna (Italy) through the application of sophisticated analysis techniques such as solid-phase microextraction gas chromatography–mass spectrometry (SPME-GC-MS) and ultra-performance liquid chromatography–mass spectrometry (UPLC-MS). The obtained data highlighted the presence of a high number of volatile and non-volatile compounds, thus allowing a comparative evaluation of the different samples. Furthermore, an in-depth statistical survey by principal component analysis (PCA) with heat maps, hierarchical cluster analysis (HCA), and partial least squares discriminant analysis (PLS-DA-VIP) was conducted to consider any correlations between the investigated cultivars. The findings of this study may help to provide more information on the C. sativa inflorescences and prove useful for potential applications of their metabolites in scientific research.

Keywords: cannabinoids, non-cannabinoids, volatile and non-volatile compounds, chromatographic analyses, multivariate statistical analysis

Introduction

Cannabis sativa L. (Cannabaceae) was one of the first non-food crops to be cultivated. It is an annual flowering herbaceous plant native to temperate central Asia, where its use seems to date back to around 4500 B.C. [1,2] This species is found in different habitats ranging from sea level to the temperate and alpine foothills of the Himalayas. [3] Its domestication probably occurred independently in several centers of East Asia in early Neolithic times. [4] Around 1000 B.C., it rapidly spread throughout Asia and Europe following the migration of nomads and the movements of traders. [2,5] C. sativa has a long history as a medicinal plant, used, for example, in traditional Tibetan and Ayurvedic medicine, as well as in social and religious rituals. The strong, mildew-resistant fiber has long been used by humans in the construction of ropes and sails. In China, the seeds are still commonly eaten roasted or raw. [6,7] In recent years, renewed interest in the therapeutic effects of C. sativa has led to legitimate medicinal use through clinical studies demonstrating its efficacy. [8] Moreover, C. sativa is currently an agricultural commodity grown to be used in the production of foods and beverages, nutritional supplements, cosmetics and personal care products, textiles, paper, insulation materials, and other manufactured goods. [9,10]

From a chemical point of view, C. sativa is a complex species with numerous (>500) reported secondary metabolites—both cannabinoid and non-cannabinoid constituents—obtained from all plant parts (i.e, leaves, flowers, bark, seeds, and roots). [11,12,13,14] The former are a specific chemical class found in the Cannabis genus, a group of compounds with a characteristic C21 terpenophenolic backbone and divided into several sub-classes. [15] The latter belong to various chemical classes including alkaloids, flavonoids, non-cannabinoid phenols, terpenes, and more. [12,15,16,17] While the stems provide cellulosic and woody fibers, and the seeds are exploited in the feed and food industry for their high content of fatty acids and proteins, the leaves and inflorescences of C. sativa are a rich source of phytochemicals. [18]

In this work, the inflorescences of five organic C. sativa commercial cultivars were chemically investigated. In particular, V1 CBD, Banana Hybrid, Green Poison, Candy BUD, and Gorilla CBD (Figure 1) were analyzed, all with a high cannabidiol content and characterized by medium/large sized buds that are compact and containing a good amount of resin. The samples were obtained from greenhouse crops located in the Tuscan-Emilian Apennines (Italy), where the entire processing cycle is carried out strictly by hand and the tanning process is particularly long in order to obtain and enhance their unique taste. In detail, Green Poison gives off an unusual smell due to contrasting sugary and bitter aromas, leaving a very pleasant aftertaste. As the name suggests, Banana Hybrid releases sweet banana fragrances. Both V1 CBD and Candy BUD have a dense and particularly fragrant smell, with fruity notes reminiscent of citrus fruits. Gorilla CBD smells of grass and pine. [19] With the aim of providing a detailed description of their chemical composition, we applied solid-phase microextraction gas chromatography–mass spectrometry (SPME-GC-MS) and ultra-performance liquid chromatography–mass spectrometry (UPLC-MS) techniques. The findings were useful for carrying out a comparative assessment also thanks to a sophisticated statistical survey that highlighted qualitative and quantitative differences in volatile and non-volatile chemical profiles.

Fig. 1 Inflorescences from the five different Cannabis sativa L. cultivars in this study

Notes

This presentation is faithful to the original, with only a few minor changes to presentation. Some grammar and punctuation was cleaned up to improve readability. In some cases important information was missing from the references, and that information was added.