Table 2.
AH-7921 and metabolites in the authentic urine forensic case specimen analyzed without and after glucuronidase hydrolysis: Retention time, accurate mass protonated molecule m/z, elemental composition, diagnostic product ions, mass error calculated for the protonated molecule [MH+] and MS peak areas of all metabolites previously identified after hepatocyte incubation as well as three additional metabolites that were among the 10 most abundant metabolites
| Peak ID | Metabolic reaction | RT (min) | Protonated molecule [MH+] (m/z) | Mass error [MH+] (ppm) | Elemental composition | Diagnostic product ions (m/z)1 | MS Area (area
rank) Non-hydrolyzed |
MS Area (area
rank) Hydrolyzed |
|---|---|---|---|---|---|---|---|---|
| a | Di-demethylation + Hydroxylation | 5.60 | 317.0820 | 0.8 | C14H18N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 1.08E+06 (6) | 9.09E+05 (5) |
| b | Di-demethylation + Hydroxylation | 6.35 | 317.0819 | 0.3 | C14H18N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 4.91E+05 (10) | 4.19E+05 (10) |
| M1 | Demethylation + Hydroxylation | 6.11 | 331.0978 | 1.1 | C15H20N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 2.80E+05 (11) | 2.50E+05 (11) |
| M2 | Di-Demethylation + Hydroxylation | 6.48 | 317.0819 | 0.3 | C14H18N2O2Cl2 | 145, 173, 190, 282, 300 | 1.44E+06 (5) | 8.86E+05 (6) |
| M3 | Di-Demethylation + Hydroxylation | 6.97 | 317.0820 | 0.5 | C14H18N2O2Cl2 | 145, 173, 190, 282, 300 | 1.66E+06 (4) | 1.35E+06 (4) |
| M4 | Demethylation + Hydroxylation + Imine Formation (artifact of Demethylation + Dihydroxylation produced by water loss) | ND | 329.0819 | ND | C15H18N2O2Cl2 | ND | ND | |
| M5 | Demethylation + Hydroxylation | 7.00 | 331.0976 | 0.4 | C15H20N2O2Cl2 | 145, 173, 190, 282, 300 | 5.18E+05 (9) | 5.20E+05 (9) |
| M6 | Demethylation + Hydroxylation | 7.36 | 331.0974 | −0.2 | C15H20N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 7.72E+05 (8) | 6.87E+05 (8) |
| M7 | Hydroxylation | 7.43 | 345.1138 | 2.1 | C16H22N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 1.17E+04 (14) | 2.54E+04 (13) |
| M8 | Hydroxylation | 7.82 | 345.1137 | 1.6 | C16H22N2O2Cl2 | 145, 173, 190, 202, 282, 300 | 5.19E+04 (12) | 1.03E+05 (12) |
| M9 | Demethylation + Hydroxylation | 8.80 | 331.0980 | 1.7 | C15H20N2O2Cl2 | 145, 173, 190, 282, 300, 313 | 1.37E+04 (13) | 9.64E+03 (14) |
| M10 | Di-Demethylation | 9.17 | 301.0872 | 0.9 | C14H18N2OCl2 | 95, 145, 173, 190, 202, 284 | 1.99E+06 (3) | 1.73E+06 (3) |
| M11 | Demethylation | 9.67 | 315.1030 | 1.4 | C15H20N2OCl2 | 95, 145, 173, 190, 202, 284 | 4.41E+06 (2) | 3.66E+06 (1) |
| Parent | 10.14 | 329.1185 | 1.0 | C16H22N2OCl2 | 95, 145, 173, 190, 202, 284 | 8.33E+05 (7) | 7.32E+05 (7) | |
| c | Di-Demethylation + Hydroxylation aglycone | 10.48 | 317.0824 | 2.0 | C14H18N2O2Cl2 | 95, 110, 145, 173, 190, 202, 225, 281, 284, 299, 317 | ND | 2.79E+06 (2) |
| M12 | Di-Demethylation + Hydroxylation + Glucuronidation | 12.31 | 493.1152 | 2.6 | C20H26N2O8Cl2 | 95, 145, 173, 190, 202, 284, 299, 317 | 4.42E+06 (1) | ND |