TABLE 2.
Overview of metabolites of 10074-G5 analyzed by HPLC-UV and LC-MS/MS
| Peak | LC-MS/MS DAD | [M+H]+ (m/z)/Cps | Product Ions | HPLC-UV | Postulated Designation a |
|---|---|---|---|---|---|
| min | min | ||||
| MS1 | 8.9 | 495/2.8 × 106 | 371, 283, 239, 195, 189, 177, 145, 133, 101, 89, 73 | Amino-O-Gluc | |
| MS2 | 9.0 | 345/1.3 × 105 | 333, 289, 277, 263, 233, 219, 189, 174, 167, 145, 101 | Amino-Acetyl | |
| MS3 | 11.1 | 495/1.5 × 106 | 302, 301, 285, 184, 177, 133, 101, 89, 73 | 6.9 (M1) b | Amino-O-Gluc |
| MS4 | 11.6 | 525/2.5 × 106 | 481, 221, 177, 133, 89, 73 | 6.9 (M1) b | Nitro-O-Gluc |
| MS5 | 12.8 | 525/1.4 × 107 | 481, 349, 332, 315, 298, 284, 273, 255, 221, 203, 133, 89, 73 | 6.9 (M1) b | Nitro-O-Gluc |
| 12.6 | |||||
| MS6 | 13.8 | 495/1.6 × 105 | 370, 327, 319, 302, 301, 285, 274, 247, 189, 184, 182, 156, 145, 113, 89 | Amino-O-Gluc | |
| MS7 | 17.2 | 495/4.0 × 105 | 469, 425, 392, 381, 343, 337, 319, 302, 301, 285, 274, 263, 249, 219, 205, 189, 182, 161, 138, 99, 89 | 7.8 (M2) c,d | Amino-O-Gluc |
| MS8 | 17.4 | 479/5.4 × 106 | 449, 437, 405, 393, 361, 349, 317, 305, 261, 243, 217, 199, 177, 133, 101, 89, 87, 73 | Amino-Gluc | |
| MS9 | 21.1 | 495/7.0 × 105 | 453, 393, 374, 343, 319, 302, 301, 288, 284, 273, 256, 242, 225, 189, 178, 169, 145 | Amino-O-Gluc | |
| MS10 | 22.0 | 525/1.5 × 106 | 349, 332, 315, 284, 273, 257, 205, 178, 151, 117, 85 | Nitro-O-Gluc | |
| MS11 | 23.7 | 525/1.5 × 106 | 493, 481, 437, 393, 349, 305, 293, 277, 261, 233, 219, 189, 175, 145, 133, 89 | Nitro-O-Gluc | |
| MS12 | 24.6 | 349/1.1 × 107 | 331, 315, 298, 284, 272, 255, 229, 197, 185, 167, 139, 128, 115, 91, 79 | 9.8 (M3) b | Nitro-OH |
| 24.3 | |||||
| MS13 | 28.0 | 303/2.9 × 106 | 286, 273, 258, 225, 209, 197, 189, 159, 153, 135, 113, 107, 99, 89, 79, 77 | Amino | |
| MS14 | 31.7 | 479/1.2 × 106 | 437, 405, 397, 355, 349, 307, 293, 263, 215, 197, 155, 135, 107, 99, 89 | Amino-Gluc | |
| MS15 | 32.2 | 303/8.0 × 105 | 285, 272, 257, 232, 205, 179, 161, 137, 133, 119, 105, 95, 91, 81, 67, 54 | Amino | |
| MS16 | 33.0 | 319/1.6 × 105 | 301, 284, 272, 257, 245, 233, 167, 145, 119, 105, 91 | Amino-OH | |
| MS17 | 34.2 | 333/6.5 × 107 | 315, 299, 282, 268, 256, 254, 242, 229, 204, 181, 167, 152, 140, 105, 78 | 12.1 (10074-G5) | Parent |
| 34.0 | |||||
| MS18 | 35.7 | 479/7.0 × 105 | 461, 429, 417, 379, 353, 321, 305, 283, 261, 239, 223, 179, 161, 158, 133, 95, 89 | Amino-Gluc | |
| MS19 | 36.8 | 345/4.8 × 105 | 303, 285, 272, 258, 255, 245, 167, 85 | Amino-Acetyl |
Designation includes: the fate of the nitro moiety, either intact as nitro or reduced to an amino moiety; and any oxidations or conjugations inferred from the data; Gluc, glucuronide.
Incubation of urine sample with β-glucuronidase results in disappearance of M1 and appearance of M3, suggesting that M1 contains a glucuronide of M3.
Incubation of urine sample with β-glucuronidase results in disappearance of M2.
M2 could not be detected by the DAD in the LC-MS/MS system. The retention time was estimated by correlating the retention times of M1, M3, and parent compound on both systems, and interpolating the expected LC-MS/MS retention time of M2 based on its retention time on the HPLC-UV system.