### Chemical Analysis of Cannabinoids in Hemp (Cannabis sativa L.) Phenotypes  
Regarding the analysis of cannabinoids, 10 different cannabinoids were identified in the 11 included phenotypes using high-performance liquid chromatography (HPLC) (Table 3). There was a significant difference in CBD-A and CBG-A cannabinoids between the CI and CII phenotypes. CII was more related to TS phenotypes. There were also significant differences in cannabinoids CBD and CBC between FS phenotypes and all other phenotypes. Figure 1: Principal component analysis (PCA) plots for phenotype averages according to essential oil components made on the first two PC scores (PC1 explained 39.44%, and PC2 explained 25.90%), with a total variance of 65.34%. CBD-A represented the highest proportion of cannabinoids in all included phenotypes, with the highest content in the four FS phenotypes (6.36-6.59%). Compared to the other phenotypes, the highest proportion of CBG-A was in phenotype CI (1.62%), followed by all FS phenotypes. Phenotype TV and CI had the highest proportion of \(\Delta\)-9-THC-A, while the highest proportions of \(\Delta\)-9-THC were in the phenotypes of FS (0.08-0.11%). The lowest proportion of \(\Delta\)-9-THC-A and \(\Delta\)-9-THC were in phenotypes TII and TIV. The CBN values were less than 0.04%, and \(\Delta\)-8-THC could not be detected in any sample. [PERSON] et al. [29] also determined the highest proportion of \(\Delta\)-9-THC-A, CBD-A, and CBG-A in clones of mother plants from chemotypes I, II, and III in an unknown variety. [PERSON] et al. [30] analyzed the cannabinoid content in 15 different hemp varieties; when comparing proportions of CBD-A, CBD, THC, and \(\Delta\)-9-THC-A in bracts of TS, their values were higher than those found in our samples, while other proportions were comparable with our data. In CS, there were higher proportions of all components, compared to the components in our CS variety, except for \(\Delta\)-9-THC-A, which was higher in our CS samples. [30]. As mentioned before, [PERSON] and [PERSON] [11] and [PERSON] et al. [12] already determined five major chemotypes based on the chemical profile of different hemp plants, whether it is marijuana or hemp type, while our classification further divides varieties in different phenotypes. Nevertheless, based on almost the same ratio between CBD and \(\Delta\) -9-THC and CBD-A and \(\Delta\)-9-THC-A, phenotypes CI and TV belong to chemotype II, while all other phenotypes were classified as chemotype III due to a high CBD-A and low \(\Delta\)-9-THC-A content. None of the phenotypes were chemotype I, which was expected since the investigation included hemp and not marijuana varieties. PCA plots for the averages of included phenotypes in all three varieties were performed according to cannabinoid content. PC1 explained 67.57%, and PC2 explained 15.70% of the variance seen in cannabinoid content. More detailed PCA plots with all analyzed variables are presented in Figure S2. There was a considerable difference between the phenotypes of FS compared to CS and TS, but we could not completely differentiate between varieties CS and TS. Nevertheless, there were greater differences between TS and CS phenotypes than between phenotypes within FS. The results are presented in Figure 3.  
Figure 2: Principal component analysis (PCA) plots for phenotypes of the Carmagnola selected variety, according to components of essential oil made on the first two PC scores (PC1 explained 42.63%, and PC2 explained 16.16%), with a total variance of 58.79%. Greater differences were observed based on essential oil proportions than proportions of cannabinoid content. Each variety was analyzed separately to identify any differences between phenotypes within varieties. As with the essential oil components, differences in cannabinoids between CS phenotypes (CI and CII) were detected, while there were no differences between the five TS and four FS phenotypes. An additional PCA was performed for the CS variety, and clear differences between the two phenotypes were observed (Figure 4).