Simple isolation method of cannabinol from Cannabis sativa to produce secondary reference standard analysis material

The simple isolation method of Cannabinol compounds from cannabis plants by maceration combined with ultrasonication assisted extraction, fractionation, separation and purification was carried out to obtain Cannabinol isolates which can be used as a reference standard analysis material. Ultrasonication was proven to shorten the extraction time, where extraction for 5 h with ultrasonication assisted for 15 min produced 5.536 % yield greater than the 24 h extraction. The results of extraction were fractionated using n-hexane and then chromatographically isolated with a column containing Silica Gel 60, with 2.5 cm diameter and a 15.3 cm height, eluted by n-hexane-ethyl acetate (90:10) solution. The fraction containing Cannabinol was purified using HPTLC preparative with eluent n-hexane-ethyl acetate (80:20). The purification of the Cannabinol isolate was further characterized by Spectrophotometer UV, FTIR, DCS, GCMS and LCMSMS and compared its profile to the reference standard of Cannabinol from Lipomed. The characterization results showed that the purified isolates had UV spectra with λ max at 219.0 nm and 284.5 nm, FTIR spectra at wave numbers 1620.21 cm-1, 1051.20 cm-1, 1581.63 cm-1, 1026.13 cm-1, 1128.36 cm-1 and 1232.51 cm-1. The DSC thermogram shows the melting point of compound is 74.36 C with 99.35 % purity, GCMS fragmentation at m/z 295, 296, 238 and 310, LCMSMS with [M + H]+ at 311.1 and MS2 at 222.95, 292.95 and 240.95 confirmed the chemical structure of the compound. The results of the characterization of pure isolates indicate that the compound produced was Cannabinol in accordance with the standard characterization profile of the reference standard of Cannabinol used.