Article Figures & Data
Figures
- Fig. 1.
Analysis of chemical constituents of goldenseal supplement. (A) Mass spectrum of goldenseal extract diluted in methanol containing 0.1% formic acid. Diluted extract was directly infused into the electrospray source and mass analyzer at a flow rate of 10 µl/min. The inset shows the area of spectrum from m/z 325 to 400. Peaks (m/z) corresponding to berberine, canadine, berberastine, canadaline, and hydrastine are numbered 1, 2, 3, 4, and 5, respectively. (B) Extracted ion chromatograms (m/z 336, 340, 352, 370, and 384) from injection of diluted goldenseal extract. Ion chromatograms were chosen based on the expected m/z for berberine, canadine, berberastine, canadaline, and hydrastine.
- Fig. 2.
Serum concentration time profile for hydrastine following administration of 2.7 g of goldenseal extract (containing 78 mg hydrastine) to healthy human volunteers. Circles are the mean ± S.E.M. (n = 11). The solid line was generated using a noncompartmental technique based on the method of residuals.
- Fig. 3.
Characteristic fragmentation pathway used in structure assignment of hydrastine metabolites. The mass spectrum was extracted from the chromatographic peak for hydrastine. Ionization was achieved using an electrospray interface operated in the positive ion mode.
- Fig. 4.
Typical LC-Time of Flight Mass Spectrometric (ToFMS) chromatograms of urine following goldenseal consumption. (A–E) represent unique extracted ion chromatograms from the same injection of urine. Proposed structure assignments for each peak are shown in Table 2.
- Fig. 5.
Proposed phase I and II metabolism of hydrastine. Specific enzymes are provided when possible, and assignment of these enzymes was made based on known metabolic reactions for noscapine, a naturally occurring structural analog of hydrastine. CYP = cyctochrome P450; MAO = monoamine oxidase; PON1 = human serum paraoxonase.
Tables
- TABLE 1
Pharmacokinetic parameters for hydrastine
Values are the mean ± standard deviation (n = 11).
Dose tmax Cmax AUC t1/2 Dose Excreted in Urine h ng/ml ng⋅h/ml h % 1.13 ± 0.31 1.2 ± 0.3 208 ± 126 495 ± 217 4.8 ± 1.4 0.16 ± 0.14 - TABLE 2
Exact mass and structure for diagnostic product ions observed in tandem mass spectrum analysis of hydrastine metabolites
m/z Metabolites with This Product Ion Structure of Product Ion 206.0812 M39 
192.1019 M20, M21, M37, M40, M41 190.0863 Hydrastine, M11, M12, M15, M18, M19 188.0706 M10, M13, M14 178.0863 M16, M27, M29, M30, M32, M33, 176.0706 M25, M34 - 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 Hydrohydrastinin162.0776 M2 C-C reduction 25.5 195.0657 195.0661 (2.0) C10H11O4 45 ± 19 6 180.0443 Meconine162.0320 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 HydrastonineM7 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: 858196-25-5307.1242 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: 58298-46-7323.0909 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 Norhydrastine321.1010 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 - TABLE 4
Predicted metabolites of canadaline detected in urine following consumption of goldenseal supplement
Identifier Reaction tR Expected Ion Measured Ion Formula Peak Area MS/MS Fragments Proposed Structure min m/z m/z m/z Canadaline 41.6 370.1654 370.1656 (0.5) C21H23NO5 470 190.0868a M43 C-O reduction, O-demethylation 35.1 358.1654 358.1651 (−0.8) C20H24NO5 177 ± 79, 311 340.1559 192.1015 M44 C-O reduction, O-demethylation, O-glucuronidation 18.9 534.1975 534.1969 (−1.1) C26H32NO11 29 ± 24, 50 358.1 192.1a M45 C-O reduction, O-demethylation, O-glucuronidation 24.8 534.1975 534.1959 (−3.0) C26H32NO11 18 ± 11, 31 358.1 354.1 178.1 M46, M47 C-O reduction 33.3 33.7 372.1811 372.1809 (−0.5) C21H26NO5 106 ± 39, 186; 136 ± 68, 225 192.1 272.1 354.1 M48 C-O reduction, O-glucuronidation 24.5, 27.0 548.2132 548.2111 (−3.8) C27H34NO11 128 ± 68, 225; 549 ± 285, 964 372.1809
M49 548.2117 (−2.7) 368.1339 192.1034 M50 C-O reduction, O-sulfation 33.3 452.1379 452.1360 (−4.5) C21H26NO8S 500 ± 268, 877 372.1799 354.1687 272.0598 192.1031 ↵a Diagnostic fragment ion. See Figure 3 for proposed fragmentation mechanism.
- TABLE 5
Area under the chromatographic peak area versus time and elimination half-life curve for each serum metabolite
Identifier m/z AUC (Peak Area × h)a Tmax t1/2 Proposed Structure h h Hydrastine 386 6435 ± 3027 1.2 ± 0.3 4.8 ± 1.4 M1 (hydrohydrastinin) 192 6966 ± 4381 1.4 ± 0.3 22 ± 18 M36 402 2745 ± 1759 1.2 ± 0.4 5 ± 2 M41 562 7797 ± 7924 3.0 ± 1 11 ± 15
In this issue
Hydrastine ADME in Humans after a Single Dose of Goldenseal
