Cannabis Extracts and Their Cytotoxic Effects on Human Erythrocytes, Fibroblasts, and Murine Melanoma

Abstract

Most of the pharmacological effects of Cannabis species have been attributed to their cannabinoid constituents, primarily Δ 9 -tetrahydrocannabinol and cannabidiol. Efficient methods of extraction, purification, and characterization of these compounds are necessary to obtain both enriched preparations and isolated compounds for therapeutic applications. In this study, forensic samples of Cannabis sativa L., Cannabaceae, were subjected to extraction, using maceration in two different solvents, methanol, and diethyl ether. These extracts were subjected to thermal decarboxylation and purification by liquid-liquid fractionation. Δ 9 -Tetrahydrocannabinol and cannabidiol concentrations from various extracts were quantified by high-performance liquid chromatography with UV detection. Extract cytotoxicity was assayed in vitro using human erythrocytes, mouse embryonic fibroblasts (NIH3T3), and murine melanoma (B16F10) cells. The B16F10 cell death mechanism was also evaluated after treatment with extract samples. The results demonstrated promising effects of various cannabinoid extracts in a murine melanoma cancer line.

Progress in the analysis of phytocannabinoids by HPLC and UPLC (or UHPLC) during 2020–2023

“…Recently, an Agilent 1200 series HPLC coupled with DAD and Poroshell column 120 SB-C 18 (150 Â 4.6 mm, 2.7 μm), eluted with a mobile phase gradient comprising 0.1% formic acid (HCOOH) in water and acetonitrile (ACN), have been used for the straightforward detection of CBD (5), CBG (8), CBN (10), Δ 9 -THC (14), and Δ 9 -THCA (15) in the aerial parts 14 (Table 1). There have been several other HPLC-based analyses of cannabinoids in the aerial parts of this plant, either using a DAD or an MS detector or in combination of both, reported in the literature [15][16][17][18][19][20][21][22][23] (Table 1). In all those studies, [15][16][17][18][19][20][21][22] the column length, T A B L E 1 HPLC and UPLC (or UHPLC) methods for the analysis of cannabinoids in Cannabis sativa L. plant samples.…”

“…There have been several other HPLC-based analyses of cannabinoids in the aerial parts of this plant, either using a DAD or an MS detector or in combination of both, reported in the literature [15][16][17][18][19][20][21][22][23] (Table 1). In all those studies, [15][16][17][18][19][20][21][22] the column length, T A B L E 1 HPLC and UPLC (or UHPLC) methods for the analysis of cannabinoids in Cannabis sativa L. plant samples. μl PDA set at 214 nm CBC (1), CBD (5), CBDA (6), CBG (8), CBGA (11), CBN (10), Δ 8 -THC (13), Δ 9 -THC (14), and 9 -THCA (15) (5), CBG (8), CBN (10), Δ 9 -THC (14), and CBC (1), CBCA (2), cannabichromevarin (CBCV), CBD (5), CBDA (6), CBDV (18), CBG (8), CBGA (9), cannabigerovarin (CBGV), CBN (10), cannabinerol (CBNR), Δ 8 -THC (13),…”

“…28,29 A simple isocratic elution with 75% ACN in water containing 0.1% HCOOH using a Luna C 18 column afforded PDA-based detection (220 nm) of several phytocannabinoids, for example, CBC (1), CBCA (2), CBCL (3), CBD (5), CBDA (6), CBDV (18), CBDVA (19), CBG (8), CBGA (9), CBN (10), Δ 8 -THC (13) Δ 9 -THC ( 14), Δ 9 -THCA (15), THCV (16), and THCVA (17). 30 A water-MeOH (both containing 0.1% HCOOH) gradient could separate CBC (1), CBCA (2), CBD (5), CBDA (6), CBDV (18), CBDVA (19), CBG (8), CBGA (9), CBN (10), Δ 9 -THC ( 14), Δ 9 -THCA (15), THCV (16), and THCVA (17) in cannabis biomass samples extracted with a pressurized solvent extraction method, 31 where an Accucore C…”

“…[1][2][3][4] Several HPLC methods have been described in the literature for the identification of cannabinoids from flowers [35][36][37][38][39][40][41][42][43][44] (Table 1). A straightforward isocratic elution with 0.1% HCOOH/0.5 mM ammonium formate in water/0.1% HCOOH/0.5 mM ammonium formate in ACN (1:3) at a flow rate of 1.5 ml/min afforded an excellent separation and PDA-based identification of CBC (1), CBCA (2), CBCL (3), CBG (8), CBN (10), CBNA (11), Δ 9 -THC (14), THCV (16), and THCVA (17) in hemp flowers. 36 This was the first study that aimed at direct comparison of the cannabinoid composition of floral extracts from unpollinated and pollinated flowers from hemp plants grown under identical conditions; it was concluded that pollination did not cause the total loss of measurable cannabinoids like CBD (5), albeit there were some differences between the phytocannabinoid composition of extracts from pollinated and nonpollinated flowers.…”

Rationalizing a prospective coupling effect of cannabinoids with the current pharmacotherapy for melanoma treatment

“…Recently, significant interest from multidisciplinary research groups worldwide has emerged, trying to utilize nonconventional remedies such as cannabinoids to treat melanoma (Carpi et al, 2017; Gruber et al, 2021; Milanez et al, 2021; Simmerman et al, 2019). Cannabinoids have been shown to manipulate various cell signaling responses that influence an immunogenic tumor's microenvironment.…”