Arboxamide structure) and m/z 257.09134 (acylium-ion right after cleavage from the C bond) have been detected. Hence, M22 was concluded to become carbonylated in the 4methyl-tetrahydropyran-moiety. 2.3.four. Tri-Hydroxylation MC2a and MC4 are RIPK1 Inhibitor manufacturer di-hydroxylated in the cumyl-moiety, as verified by detection with the fragment at m/z 151.0754. MC2a and MC4 also present a fragment at m/z 408.1918 because of water loss for the duration of fragmentation of your otherwise intact structure. Because of theMetabolites 2021, 11,10 ofobserved water loss, the place of the 1 hydroxyl group is situated at the 4-methyltetrahydropyran-moiety. MC5 was observed to become mono-hydroxylated in the cumyl-moiety, displaying the diagnostic fragment at m/z 135.0804. The added fragment at m/z 256.1081, resulting from two dehydration reactions of the di-hydroxylated 1-(tetrahydropyranyl-4methyl)-indazole-3-carboxamide structure, verifies the positions in the two other hydroxyl groups at the 4-methyl-tetrahydropyran-moiety. In-source water loss of MC5, major towards the signal of MCArt1, couldn’t be ruled out, due to the proximity of MC5 along with the observed signal of MCArt1, which also has a single hydroxyl-group in the cumyl-moiety (m/z 135.0804) but is hydroxylated and on top of that desaturated in the tetrahydropyran-moiety. As a result, MCArt1 was defined as a feasible artefact. Mono-hydroxylation at the cumyl-moiety was also observed for MC7. As for MC7, only one particular dehydration PRMT4 Inhibitor Storage & Stability reaction was detected, indicated by the fragment at m/z 274.1186. Observed fragments for MC7 indicated mono-hydroxylation at the cumyl-moiety, the indazole-core, and at the 4-methyl-tetrahydropyran-moiety. This was also confirmed by means of the derivatization experiment, as the methylated solution of MC7 was detected, presenting a diagnostic fragment at m/z 306.1448, which represents the di-hydroxylated and methylated 1-(tetrahydropyranyl-4-methyl)-indazole-3-carboxamide-moiety. MC9 is di-hydroxylated at the cumyl-moiety, as shown by the fragment at m/z 151.0754. Moreover, a fragment at m/z 258.1237 was detected, which was the dehydration product on the 1-(tetrahydropyranyl4-methyl)-indazole-3-acylium-ion, as a result indicating the place of the third hydroxyl group in the 4-methyl-tetrahydropyran-moiety. MC10 is suggested to become di-hydroxylated at the 4methyl-tetrahydropyran-moiety, but additionally mono-hydroxylated at the indazole-core. Further, an ion corresponding towards the solution of tri-hydroxylation and methylation of MC10 at m/z 440.2180 was detected after derivatization. Fragmentation of this methylated metabolite produced a diagnostic ion at m/z 322.1397, referring to the methylated tri-hydroxylated 1-(tetrahydropyranyl-4-methyl)-indazole-3-acylium-ion, and hence verifying the place of one hydroxyl group at the unsaturated indazole-region. MC11 is tri-hydroxylated in the 1(tetrahydropyranyl-4-methyl)-indazole-3-carboxamide structure, because the fragment standing for the tri-hydroxylated 1-(tetrahydropyranyl-4-methyl)-indazole-3-carboxamide-moiety (m/z 308.1241) was detected. Furthermore, this moiety made further fragments, just after one particular (m/z 290.1135), two (m/z 272.1030), and 3 dehydrations (m/z 254.0924). Derivatization didn’t lead to a decline on the MC11 signal, hence confirming the place of all three hydroxyl-groups in the unsaturated 4-methyl-tetrahydropyran-moiety. two.3.five. Mono-Hydroxylation and More Desaturation and Carbonylation MC3 is probably formed via metabolic tri-hydroxylation (MC5) and concurrent dehydrati.