TABLE 3
Chromatographic and ESI-TOF data of hydrastine and proposed metabolites in urine from healthy volunteers following oral administration of 2.7 g of goldenseal supplement (equivalent to 78 mg of hydrastine)
| Identifier | Proposed Reaction | tR | Expected Ion | Measured Ion | Formula | Peak Areaa | Hydrastine | MS/MS Fragments | Proposed Structure |
|---|---|---|---|---|---|---|---|---|---|
| min | m/z | m/z | % | m/z | |||||
| Hydrastine | Parent | 37.3 | 384.1447 | 384.1430 | C21H20NO6 | 625 ± 390 | 100 | 190.0892b |  |
| 293.0850 | |||||||||
| 323.0825 | |||||||||
| M1 | C-C reduction | 14.2 | 192.1024 | 192.1018 (+1.0) | C11H13NO2 | 261 ± 135 | 34 | 177.0787 |  Hydrohydrastinin |
| 162.0776 | |||||||||
| M2 | C-C reduction | 25.5 | 195.0657 | 195.0661 (2.0) | C10H11O4 | 45 ± 19 | 6 | 180.0443 |  Meconine |
| 162.0320 | |||||||||
| M3 | C-C reduction, dehydrogenation/C-C reduction, C-O reduction, aromatization | 15.4, 15.8 | 190.0868 | 190.0866 (−1.0), 190.0879 (+5.7) | C11H12NO2 | 1167 ± 509; 778 ± 312 | 151 | M3: 175.0633 |  |
| 147.0677 | |||||||||
| 154.0508 | |||||||||
| 132.0365 | |||||||||
| M4 | 101 | M4: 175.0626 | |||||||
| 160.0755 | |||||||||
| 149.0581 | |||||||||
| 132.0365 | |||||||||
| M5 | C-C reduction, aromatization | 15.6 | 188.0712 | 188.0714 (+1.0) | C11H10NO2 | 3860 ± 1442 | 500 | 130.0555, 160.0676 |  Hydrastonine |
| M7 | C-C reduction, N-demethylation and N-sulfation | 11.9 | 258.0436 | 258.0434 (−0.8) | C10H12NO6S | 137 ± 72 | 18 | 178.0868 |  |
| M9 | C-C reduction, N-demethylation, hydroxylation and O-glucuronidation | 22.4 | 370.1138 | 370.1138 (0) | C16H20NO9 | 99 ± 77 | 13 | 194.0812 |  |
| M10 | C-C dehydrogenation | 30.8 | 382.1291 | 382.1278 (−3.4) | C21H20NO6 | 74 ± 42 | 10 | 334.0744 |  |
| 322.1118 | |||||||||
| 292.1011 | |||||||||
| 278.0926 | |||||||||
| 188.0712b | |||||||||
| M11 | Lactone hydrolysis, alcohol dehydrogenation | 27.6 | 400.1396 | 400.1388 | C21H22NO7 (−2.0) | 186 ± 59 | 24 | 382.1301 |  |
| 190.0866b | |||||||||
| M12 | Hydroxylation | 37.3 | 400.1396 | 400.1402 | C21H22NO7 (+1.5) | 304 ± 157 | 39 | 190.0866b |  |
| 311.0924 | |||||||||
| 323.0917 | |||||||||
| 335.0938 | |||||||||
| 353.1053 | |||||||||
| M13,14 | Lactone hydrolysis, dehydrogenation, acyl glucuronidation | 27.2, 28.8 | 576.1717 | 576.1706 (−1.9), 576.1705 (−2.1) | C27H30NO13 | 72 ± 50 | 9 | M13: |  |
| 382.1235 | |||||||||
| 188.0721b | |||||||||
| M14: 382.1181 | |||||||||
| 188.0697b | |||||||||
| M15 | C-C dehydrogenation | 32.4 | 382.1291 | 382.1289 (−0.5) | C21H20NO6 | 154 ± 98 | 20 | 322.1089 |  CAS: 858196-25-5 |
| 307.1242 | |||||||||
| 292.0983 | |||||||||
| 278.0943 | |||||||||
| 280.1007 | |||||||||
| 190.0882b | |||||||||
| M16 | C-O reduction, O-demethylation | 21.4 | 372.1447 | 372.1437 (−2.7) | C20H22NO6 | 178 ± 100 | 23 | 354.1255 |  CAS: 58298-46-7 |
| 323.0909 | |||||||||
| 311.0941 | |||||||||
| 178.0859b | |||||||||
| M18 | O-demethylation, O-sulfation | 27.4 | 450.0860 | 450.0851 (−1.8) | C20H20NO9S | 73 ± 70 | 9 | 370.1304 |  |
| 309.0805 | |||||||||
| 190.0887b | |||||||||
| M19 | O-demethylation, O-glucuronidation | 19.9 | 546.1612 | 546.1607 (−0.9) | C26H28NO12 | 508 ± 359 | 66 | 370.1288 |  |
| 309.0797 | |||||||||
| 190.0874b | |||||||||
| M20 | C-O reduction, O-demethylation, O-glucuronidation | 12.1 | 548.1768 | 548.1740 (−5.1) | C26H30NO12 | 355 ± 211 | 46 | 372.1447 |  |
| 368.1266 | |||||||||
| 311.0951 | |||||||||
| 294.1451 | |||||||||
| 192.1035b | |||||||||
| M21 | C-O reduction, O-demethylation, O-glucuronidation | 12.5 | 548.1768 | 548.1758 (−1.8) | C26H30NO12 | 533 ± 316 | 69 | 372.1425 | |
| 323.0960 | |||||||||
| 311.0886 | |||||||||
| 192.1035b | |||||||||
| 165.0651 | |||||||||
| 144.0820 | |||||||||
| M22 | C-O reduction, O-demethylation, O-sulfation | 18.8 | 452.1016 | 452.1016 (0) | C20H22NO9S | 117 ± 81 | 15 | 372.1456 |  |
| 323.0952 | |||||||||
| 311.0924 | |||||||||
| 192.1018b | |||||||||
| M25 | N-demethylation, O-demethylation, O-glucuronidation | 19.3 | 532.1455 | 532.1452 (−0.6) | C25H26NO12 | 220 ± 138 | 29 | 514.1329 |  |
| 356.1135 | |||||||||
| 338.1030 | |||||||||
| 176.0707b | |||||||||
| M 27 | C-O reduction, N-demethylation, O-demethylation, O-sulfation | 16.9 | 438.0860 | 438.0851 (−1.8) | C19H20NO9S | 145 ± 88 | 19 | 358.1266 |  |
| 258.0418 | |||||||||
| 297.0751 | |||||||||
| 178.0871b | |||||||||
| M 29 | C-O reduction, N-demethylation, O-demethylation, O-sulfation | 18.6 | 438.0860 | 438.0867 (1.8) | C19H20NO9S | 48 ± 26 | 6 | 358.1269 | |
| 178.0868b | |||||||||
| M 30 | C-O reduction, N-demethylation, O-demethylation, O-glucuronidation | 12.7 | 534.1612 | 534.1601 (−2.1) | C25H28NO12 | 74 ± 73 | 10 | 516.1489 |  |
| 358.1084 | |||||||||
| 340.1005 | |||||||||
| 178.0872b | |||||||||
| M 31 | N-demethylation, C-O reduction | 31.2 | 372.1447 | 372.1447 (0) | C20H22NO6 | 280 ± 128 | 36 | 354.1348 |  |
| 178.0875b | |||||||||
| 323.1196 | |||||||||
| M 32 | N-demethylation, C-O reduction, O-glucuronidation | 16.7 | 548.1768 | 548.1755 (−2.4) | C26H30NO12 | 579 ± 489 240 ± 108 | 75 | M32: 372.1442 |  |
| 354.1183 | |||||||||
| 178.0866b | |||||||||
| M 33 | 17.3 | 548.1753 (−2.7) | 31 | M33: 372.1435 | |||||
| 354.1173 | |||||||||
| 178.0865b | |||||||||
| M34 | N-demethylation | 34.3 | 370.1290 | 370.1300 | C20H20NO6 | 176.0796b |  Norhydrastine | ||
| 321.1010 | |||||||||
| 352.1186 | |||||||||
| M 36 | Lactone hydrolysis | 34.0 | 402.1553 | 402.1543 (−2.5) | C21H24NO7 | 329 ± 209 | 28 | 384.1447 |  |
| 366.1384 | |||||||||
| 353.1024 | |||||||||
| 335.0919 | |||||||||
| 323.0924 | |||||||||
| M 37 | Lactone hydrolysis, C-O reduction, O-glucuronidation | 15.9 | 580.2030 | 580.2019 (−1.9) | C27H34NO13 | 768 ± 504 | 100 | 562.1642 |  |
| 404.1642 | |||||||||
| 386.1659 | |||||||||
| 368.1326 | |||||||||
| 192.1000b | |||||||||
| M 39 | Hydroxylation, O-glucuronidation | 27.7 | 576.1717 | 576.1705 (−2.1) | C27H30NO13 | 235 ± 125 | 38 | 400.1353 |  |
| 206.0389b | |||||||||
| 451.2171 | |||||||||
| 467.1917 | |||||||||
| M 40 | C-O reduction | 25.8 | 386.1604 | 386.1605 (0.3) | C21H24NO6 | 482 ± 246 | 62 | 368.1318 |  |
| 337.1096 | |||||||||
| 325.1057 | |||||||||
| 192.1013b | |||||||||
| M 41 | C-O reduction, O-glucuronidation | 18.4 | 562.1925 | 562.1921 (−0.4) | C27H32NO12 | 2389 ± 1145 | 309 | 386.1584 |  |
| 368.1327 | |||||||||
| 325.1041 | |||||||||
| 192.1028b |