Supplementary Materials: Detection of 191 Taxifolin Metabolites and Their Distribution in Rats Using HPLC-ESI-IT-TOF-MS n

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1 S1 of S26 Supplementary Materials: Detection of 191 Taxifolin Metabolites and Their Distribution in Rats Using HPLC-ESI-IT-TF-MS n Ping Yang, Feng Xu, Hong-Fu Li, Yi-Wang, Feng-Chun Li, Ming-Ying Shang, Guang-Xue Liu, Xuan Wang, Shao-Qing Cai

2 S2 of S26 Table S1. Retention time (tr), HRMS n data, molecular formula and identification of taxifolin and its metabolites in rats by HPLC-DAD-ESI-IT-TF-MS n. tr(min) Formula Meas. m/z Identification ther Ions TAX C15H Taxifolin(parent compound) [M + H] + : M C15H Taxifolin isomer 1 M C15H Taxifolin isomer 2 M C15H1210S Taxifolin sulphate 1 M C15H1210S Taxifolin sulphate 2 M C15H1210S Taxifolin sulphate 3 M C15H1210S Taxifolin sulphate 4 M C15H1210S Taxifolin sulphate 5 M C15H1210S Taxifolin sulphate 6 [M + H] + : [2M H] : [M + H] + : [2M H] : [M + HCH H] : [M + HCH H] : M C15H1210S Taxifolin sulphate 7 [M + H] + : M C15H1210S Taxifolin sulphate 8 [M + H] + : M C15H1210S Taxifolin sulphate 9 [M + H] + : MS 2 [ ]: (10), (26.25), (3.33), (12.87), (15.76), (8.65); MS 3 [ ]: (49.03), (14.55), (41.19), (10) MS 2 [ ]: (10), (18.49), (11.8), (10.39), (8.51); MS 3 [ ]: (100), (35.54), (94.08) MS 2 [ ]: (10), (33.23), (5.75), (9.38), (7.83); MS 3 [ ]: (60.16), (10) MS 2 [ ]: (5.67), (10), (17.89), (23.67), (3.4); MS 3 [ ]: (10), (31.44), (46.01), (85.87) MS 2 [ ]: (12.71), (10), (14.26), (10.69), (1.69); MS 3 [ ]: (65.85), (27.1), (41.93), (10) MS 2 [ ]: (11.06), (10), (1.76), (11.2), (4.11), (1.39); MS 3 [ ]: (12.25), (24.74), (18.5), (10) MS 2 [ ]: (9.45), (10), (17.35), (16.0), (4.14); MS 3 [ ]: (16.81), (16.81), (39.63), (10) MS 2 [ ]: (0.69), (10), (16.27), (5.3), (9.78) MS 2 [ ]: (11.05), (10), (22.65), (10.85), (17.28), (1.0); MS 3 [ ]: (10), (36.07), (43.17) MS 2 [ ]: (15.4), (10), (20.79), (5.02), (14.67); MS 3 [ ]: (14.5), (44.76), (22.02), (45.3), (10) MS 2 [ ]: (11.96), (10), (20.93), (15.46), (2.32); MS 3 [ ]: (47.49), (61.46), (10), (5.86), (1.41) MS 2 [ ]: (9.17), (10), (21.31), (14.8), (16.18); MS 3 [ ]: (20.19), (17.67), (57.12), (10)

3 S3 of S26 tr(min) Formula Meas. m/z Identification ther Ions M C15H1213S Taxifolin disulphate 1 [M + CH3CH H] : M C15H1213S Taxifolin disulphate 2 [M + 2H2 H] : M C15H1213S Taxifolin disulphate 3 M C15H1213S Taxifolin disulphate 4 M C20H19N13 S Taxifolin sulphate and pyroglutamic acid conjugate M C21H Taxifolin glucuronide 1 M C21H Taxifolin glucuronide 2 [M + HCH H] : [M + CH3CH H] : [M + H] + : [M + CH3CH H] : [M + HCH H] : M C21H Taxifolin glucuronide 3 [M + H] + : M C21H Taxifolin glucuronide 4 [M + H] + : M C21H Taxifolin glucuronide 5 [M + H] + : M C21H Taxifolin glucuronide 6 [M + H] + : M C21H Taxifolin glucuronide 7 [M + H] + : MS 2 [ ]: (10), (3.04), (1.52); MS 3 [ ]: (10), (11.61), (2.58), (17.08), (8.59) MS 2 [ ]: (10), (2.89), (8.47), (4.65); MS 3 [ ]: (10), (5.91), (11.25), (14), (10.13) MS 2 [ ]: (10), (1.14), (1.69); MS 3 [ ]: (7.04), (10), (12.38), (3.59), (3.59) MS 2 [ ]: (10), (3.2), (5.53); MS 3 [ ]: (5.89), (10), (38.26), (22.51), (4.89) MS 2 [ ]: (28.19), (22.72), (10), (28.19), (71.79), (13.11), (13.8); MS 3 [ ]: (18.05), (10), (22.65), (31.86), (18.05) MS 2 [ ]: (21.29), (10), (42.59), (15.29), (12.24) MS 2 [ ]: (24.22), (10), (10.69), (11.32), (10.89); MS 3 [ ]: (41.83), (10) MS 2 [ ]: (17.51), (10), (21.38), (16.27), (2.08); MS 3 [ ]: (14.07), (14.07), (21.1), (10), (45.58) MS 2 [ ]: (32.82), (10), (13.88), (17.26), (6.27); MS 3 [ ]: (72.43), (10), (58.38) MS 2 [ ]: (28.1), (10), (10.52), (4.3), (1.54), (3.09) MS 2 [ ]: (2.13), (27.43), (10), (6.27), (11.87), (2.48); MS 3 [ ]: (10), (85.25), (57.92) MS 2 [ ]: (10), (4.72), (5.84), (22.56), (4.72), (34.32); MS 3 [ ]: (10), (8) M C21H Taxifolin glucuronide 8 [M + H] + : N M C21H Taxifolin glucuronide 9 [M + H] + : MS 2 [ ]: (3.1), (10), (6.68), (21.39), (8.25), (14.67); MS 3 [ ]: (5), (10), (74.51), (62.25)

4 S4 of S26 tr(min) Formula Meas. m/z Identification ther Ions M C21H2016S Taxifolin glucuronide sulphate 1 [M + K] + : M C21H2016S Taxifolin glucuronide sulphate 2 [M + 2H2 H] : M C21H2016S Taxifolin glucuronide sulphate 3 [M + 2H2 H] : M C21H2016S Taxifolin glucuronide sulphate 4 M C21H2016S Taxifolin glucuronide sulphate 5 M C21H2016S Taxifolin glucuronide sulphate 6 M C21H2016S Taxifolin glucuronide sulphate 7 [M + HCH H] : [M + CH3HCH H] : [M + K] + : M C16H Methyltaxifolin [M + H] + : M C16H Methyltaxifolin [M + H] + : M C16H Methyltaxifolin [M + H] + : M C16H Methyltaxifolin [M + H] + : M C16H1410S Methyl taxifolin sulphate 1 [M + H] + : M C16H1410S Methyl taxifolin sulphate 2 [M + H] + : MS 2 [ ]: (10), (29.1), (86.56), (19.52), (19.52) MS 2 [ ]: (3.82), (88.44), (38.47), (10), (5.76) MS 2 [ ]: (1.96), (93.41), (50.31), (10), (10.52), (4.95) MS 2 [ ]: (2.9), (10), (49.6), (88.61), (15.36), (2.03); MS 3 [ ]: (10), (4.04), (15.28), (6.1), (31.64), (17.22) MS 2 [ ]: (10), (18.56), (89.58), (2.6), (1.96); MS 3 [ ]: (10), (7.03), (10.55), (10.55) MS 2 [ ]: (10), (39.46), (91.03), (1.65), (18.71), (6.0) MS 2 [ ]: (10), (23.09), (71.61), (10.1), (1.82), (1.21) MS 2 [ ]: (4.3), (10), (1.81), (6.36), (7.17), 23596(6.54), (7.15), (6.28); MS 3 [ ]: (20.25), (16.06), 23642(38.12), (10), (17.29) MS 2 [ ]: (4.08), (10), (3.09), 23592(6.3), (7.56); MS 3 [ ]: (13.72), (24.58), (15.28), (10), (16.05) MS 2 [ ]: (16.86), (51.55), (11.55), 23602(33.52), (23), (11.55), (6.53), (13.14), (10), (75.55); MS 3 [ ]: (10), (9.13) MS 2 [ ]: (46.64), (65.79), (29.14), (51.89), 24466(20.24), 23559(27.86), (10), (53.06); MS 3 [ ]: (66.23), 23723(10), (10) MS 2 [ ]: (95.45), (1.59), (10), (8.37), (2.21), (3.5); MS 3 [ ]: (21.25), (7.08), (7.08), (10), (7.08) MS 2 [ ]: (97.68), (2.38), (10), (3.37), (7.19), (5.98); MS 3 [ ]:

5 S5 of S26 tr(min) Formula Meas. m/z Identification ther Ions M C16H1410S Methyl taxifolin sulphate 3 M C16H1410S Methyl taxifolin sulphate 4 [M + HCH H] : M C16H1410S Methyl taxifolin sulphate 5 [M + H] + : M C16H1410S Methyl taxifolin sulphate 6 [M + H] + : M C16H1410S Methyl taxifolin sulphate 7 [M + H] + : M C16H1410S Methyl taxifolin sulphate 8 [M + H] + : M C16H1410S Methyl taxifolin sulphate 9 [M + H] + : M C16H1410S Methyl taxifolin sulphate 10 [M + CH3HCH H] : M C22H Methyl taxifolin glucuronide 1 [M + H] + : M C22H Methyl taxifolin glucuronide 2 [M + H] + : M C22H Methyl taxifolin glucuronide 3 [M + H] + : (4.97), (5.81), 23636(4.13), (10), (3.29), (3.29) MS 2 [ ]: (10), (10), (4.17), (2.09), (5.55) MS 2 [ ]: (10), (1.27), (83.29), (8.94), (3.43), (1.27); MS 3 [ ]: (10.55), (10), (21.01), (8.44), (8.44) MS 2 [ ]: (10), (4.32), (97.36), (3.75), (5.08), (0.52); MS 3 [ ]: (2.69), (10), (2.69), (19.53), (11.28), (10.3) MS 2 [ ]: (95.02), (1.12), (10), (10.29), (4.86); MS 3 [ ]: 23637(28.09), (22.55), (10) MS 2 [ ]: (76.6), (1.14), (10), (7.7), (13.71), (7.5), 23565(7.61), (5.16); MS 3 [ ]: (10.84), 23651(7.13), (10) MS 2 [ ]: (86.98), (2.81), (10), (2.81), (9.12), (4.65), (2.13), (2.56); MS 3 [ ]: (11.83), 23538(20.75), (10), (11.83) MS 2 [ ]: (81.59), (4.06), (10), (7.76), (4.4), (6.64), (3.27); MS 3 [ ]: (8.95), (4.49), 23579(28.57), (10), (3.62) MS 2 [ ]: (97.99), (38.06), (76.49), (18.77), (13.35), (10), (26.72), (4.48); MS 3 [ ]: (10), (18.37), 15244(27.21) MS 2 [ ]: (10), (1.28), (65.83), (5.82), (3.87), 23608(4.52); MS 3 [ ]: (10), (29.98) MS 2 [ ]: (10), (2.98), (86.87), (2.96), (2.22), (1.86); MS 3 [ ]: (10), 23579(20.19), (29.9) MS 2 [ ]: (91.34), (2.65), (10), (9.18), (5.73), (3.78), (5.88),

6 S6 of S26 tr(min) Formula Meas. m/z Identification ther Ions 23609(4.53); MS 3 [ ]: (11.46), 23493(9.55), (10), (11.46) M C22H Methyl taxifolin glucuronide 4 [M + H] + : MS 2 [ ]: (10), (16.32), (98.02), (4.15), (10.31), (2.41); MS 3 [ ]: (10), (27.04), (27.04) M C22H Methyl taxifolin glucuronide 5 [M + H] + : MS 2 [ ]: (17.95), (7.65), (19.27), (41.77), (39.97), (10), (10.41), (7.17); MS 3 [ ]: (10) M C22H Methyl taxifolin glucuronide 6 [M + H] + : MS 2 [ ]: (55.37), (9.98), (10), (5.43), (4.2), 23615(5.98), (5.63); MS 3 [ ]: (57.92), (57.92), (57.92), (10) M C22H Methyl taxifolin glucuronide 7 [M + H] + : MS 2 [ ]: (1.25), (2.45), (60.45), (18.79), (10), (2.15), (1.86), (5.52), (8.17), (1.25) M C22H Methyl taxifolin glucuronide 8 [M + H] + : MS 2 [ ]: (9.65), (7.22), (10), (8.43), (65.09), (55.48), (93.96), (13.26) M C22H Methyl taxifolin glucuronide 9 [M + H] + : N M C22H2216S M C22H2216S M C21H21N M C21H21N M C16H1411S M C16H1411S M C16H1411S Methyl taxifolin glucuronide sulphate 1 Methyl taxifolin glucuronide sulphate 2 Methyl taxifolin pyroglutamic acid conjugate 1 Methyl taxifolin pyroglutamic acid conjugate 2 Hydroxylated methyl taxifolin sulphate 1 Hydroxylated methyl taxifolin sulphate 2 Hydroxylated methyl taxifolin sulphate 3 MS 2 [ ]: (79.52), (10), (25), (3.96), (2.82); MS 3 [ ]: (45.61), (45.69), (10), (14.01), (11.94), (8.11), (11.94) MS 2 [ ]: (95.06), (10), (27.89), (15.21), (4.76), (3.57); MS 3 [ ]: (66.23), (66.23), (10) [M + H] + : N [M + H] + : MS 2 [ ]: (3.05), (27.21), (10), (4.57) MS 2 [413.02]: (66.16), (6.39), (10), (8.51), (36.67), (19.97); MS 3 [ ]: (8.8), (14.05), (66.56), (7.09), (10) N MS 2 [ ]: (4.33), (10), (18.88), (25.58), (6.5), (7.26), (5.93), (25), (18.85); MS 3 [ ]: 30565(21.22),

7 S7 of S26 tr(min) Formula Meas. m/z Identification ther Ions M C16H1411S M C16H M C16H M C22H M C22H M C22H M C22H Hydroxylated methyl taxifolin sulphate 4 Methylated and dihydroxylated taxifolin 1 Methylated and dihydroxylated taxifolin 2 Methylated and dihydroxylated taxifolin glucuronide 1 Methylated and dihydroxylated taxifolin glucuronide 2 Methylated and dihydroxylated taxifolin glucuronide 3 Methylated and dihydroxylated taxifolin glucuronide 4 [M + H] + : [M + K] + : [M + K] + : [M + K] + : [M + K] + : M C15H Quercetin [M + H] + : M C15H1010S Quercetin-5--sulphate [M + H] + : M C15H1010S Quercetin-7--sulphate [M + H] + : M C15H1010S Quercetin- 4 --sulphate [M + H] + : (68.37), (94.29), (21.22), (10), (21.22) MS 2 [ ]: (6.35), (10), (9.05), 29406(26.17), (12.67), (3.65), (5.47), (7.23), (6.35), (9.12), (3.65), (5.47), (3.65) MS 2 [ ]: (45.13), (5.7), (5.7), (10), (38.12), (15.5), (7.12), (97.97), (5.7) MS 2 [ ]: (38.24), (13.9), (13.9), (10), (13.9), (31.02), (13.9), (79.28), (13.9), (20.86) MS 2 [ ]: (17.05), (4.38), (88.77), (10.64), 30237(44.17), (10), (4.38), (6.45), (79.82), (8.53), (6.45) MS 2 [ ]: (19.31), (10), (1.95), (19.8), 30266(31.91), (71.56), (3.87), (65.13), (2.89), (2.91); MS 3 [ ]: (29.61), 30237(58.77), (35.54), (10) MS 2 [ ]: (23.98), (10), (22.62), 30256(49.86), (62.42), (52.95); MS 3 [ ]: 30247(36.78), (20.83), (10), (6.38) MS 2 [ ]: (37.32), (10), (12.54), 30267(46.8), (75.71), (67.54), (5.14); MS 3 [ ]: (9.56), 30309(25.46), (59.2), (10), (6.41) MS 2 [ ]: (1.27), (4.34), (5.52), (1.1), (3.45), (2.73), (3.38), (46.63), (10), (3.27), (6.98) MS 2 [ ]: (10), (2.12), (2.11); MS 3 [ ]: (7.24), (9.5), (18.99), (4.84), (7.29), (32.68), (12.95), (10), (15.59) MS 2 [ ]: (10); MS 3 [ ]: (10), (72.13) MS 2 [ ]: (10), (7.84), (5.4); MS 3 [ ]: (7.11), (10.67), (14.22), (26.53), (10)

8 S8 of S26 tr(min) Formula Meas. m/z Identification ther Ions M C15H1010S Quercetin-3 --sulphate [M + H] + : M C15H1010S Quercetin-3--sulphate M C21H Quercetin glucuronide [M + H] + : M C21H1816S Quercetin glucuronide sulphate 1 [M + Na] + : M C21H1816S Quercetin glucuronide sulphate 2 [M + 2H2 H] : M C21H1816S Quercetin glucuronide sulphate 3 [M + HCH H] : M C16H Isorhamnetin [M + H] + : M C16H1210S Isorhamnetin-5--sulphate [M + H] + : M C16H1210S Isorhamnetin-7--sulphate [M + H] + : M C16H1210S Isorhamnetin-3--sulphate [M + H] + : M C16H1210S Isorhamnetin-4 --sulphate M C16H1213S Isorhamnetin disulphate [M + H] + : [M + HCH H] : [M + HCH H] : M C22H Isorhamnetin-4 --glucuronide [M + H] + : M C22H Isorhamnetin-7--glucuronide [M + H] + : M C22H2016S Isorhamnetin glucuronide sulphate 1 [M + H] + : M C22H2016S Isorhamnetin glucuronide sulphate 2 [M + H] + : MS 2 [ ]: (10), (10.35), (5.43); MS 3 [ ]: (11.66), (9.83), (7.62), (69.17), (10), (1.09), (4.38) MS 2 [ ]: (10), 30739(4.11), (24.98), (22.85) MS 2 [ ]: (5.11), (10), (1.86), (17.1), (2.79), (1.86) MS 2 [ ]: (3.6), (10), (1.64) MS 3 [ ]: (62.02), (5), (10) MS 2 [ ]: (5.04), (0.67), (10), (2.62); MS 3 [ ]: (86.98), (10) MS 2 [ ]: (3.47), (10), (0.91) MS 2 [ ]: (30.16), 30274(10), (5.39), (3.37), (7.64); MS 3 [ ]: (10), (8.31), (14.39), (16.62), (33.01), (24.7), (8.31) MS 2 [ ]: (10), 30257(9.79), (2.71), (1.35), (0.77); MS 3 [ ]: 30212(10), (6.56), (4.43), (8.78), (4.43) MS 2 [ ]: (10), 30279(10.27), (0.94), (4.64); MS 3 [ ]: 30297(10), (7.32), (9.19), (25.84), (67.22), (7.32) MS 2 [ ]: (10), (1.79), 30266(17.54), (0.13); MS 3 [ ]: (42.99), 30265(10), (4.58), (2.67), (8.71) MS 2 [ ]: (10), 30299(33.72), (5.5), (3.69), (7.31), (3.69), (3.69), (3.69); MS 3 [ ]: 30352(10), (11.01) MS 2 [ ]: (83.92), (10); MS 3 [ ]: 30299(10) MS 2 [ ]: (10), 30276(34.63), (1.74); MS 3 [ ]: 30311(10), (7.71) MS 2 [ ]: (10), (7.02), 30301(31.77); MS 3 [ ]: 30401(10) MS 2 [ ]: (1.6), (10), 30234(26.2), (1.9); MS 3 [ ]: 30288(10) MS 2 [ ]: (5.07), (0.3), (10), 30282(31.47), (1.94); MS 3 [ ]: 30252(10),

9 S9 of S26 tr(min) Formula Meas. m/z Identification ther Ions (4.93), (1.66), (2.21), (1.66), (3.29) M C22H2016S Isorhamnetin glucuronide sulphate 3 MS 2 [ ]: (14.58), (18.16), (10) M C22H2016S Isorhamnetin glucuronide sulphate 4 MS 2 [ ]: (4.72), (1.58), (10), 3024(14.53); MS 3 [ ]: (24.92), 30268(10), (16.83), (20.87), (41.75) M C15H1011S Hydroxylated quercetin sulphate 1 [M + H] + : MS 2 [ ]: (41.74), (28.16), (17.99), (6.24), (10.43), (7.49), (4.16), (24.59), (10), (3.31), (1.66), (32.05) M C15H1011S Hydroxylated quercetin sulphate 2 [M + H] + : MS 2 [ ]: (54.7), (24.85), (14.75), (4.86), (17.0), (3.65), (26.94), (10), (4.86), (24), (3.65) M C15H1011S Hydroxylated quercetin sulphate 3 [M + H] + : MS 2 [ ]: (74.16), 30729(4.26), (7.41), (10.56), (7.41), (25.43), (8.44), (6.39), (5.32), (7.41), (32.76), (10), (40.13) M C21H M C21H M C16H1211S M C16H1211S M C16H1211S M10 0 M10 1 M C16H1211S C22H C22H Hydroxylated quercetin glucuronide 1 Hydroxylated quercetin glucuronide 2 Hydroxylated isorhamnetin sulphate 1 Hydroxylated isorhamnetin sulphate 2 Hydroxylated isorhamnetin sulphate 3 Hydroxylated isorhamnetin sulphate 4 Hydroxylated isorhamnetin glucuronide 1 Hydroxylated isorhamnetin glucuronide 2 [M + HCH H] : [M + H] + : [M + H] + : [M + H] + : MS 2 [ ]: (2.42), (10), (26.78), (32.73), (38.18), (88.23); MS 3 [ ]: (66.67), (10), (66.67) MS 2 [ ]: (90.58), (21.56), (18.18), (33.94), (10), (9.09) MS 2 [ ]: (11.57), (78.23), (10), (3.17), (28.07), (14.06), (2.64); MS 3 [ ]: (15.89), (10), 26211(35.83) MS 2 [ ]: (73.82), (8.54), (10), (2.17), (35.38), (2.17), (2.17); MS 3 [ ]: (10), (23.63), (13.69) MS 2 [ ]: (18.28), (79.96), (2.34), (10), (7.45), (44.52), (2.34); MS 3 [ ]: (10), 26189(34.75), (95.74) MS 2 [ ]: (86.27), (50.37), (10), (3.73), (3.1), (2.47), (2.47), (3.73), (2.47); MS 3 [ ]: (50.48), (50.48), (10) MS 2 [ ]: (60.2), (10), (33.72), (27.21); MS 3 [ ]: (10), 26342(66.23), (83.12) [M + H] + : N

10 S10 of S26 M10 3 M10 4 M10 5 M10 6 M10 7 M10 8 M10 9 M11 0 M11 1 M11 2 M11 3 M11 4 M11 5 M11 6 tr(min) Formula Meas. m/z Identification ther Ions C22H Hydroxylated isorhamnetin glucuronide 3 [M + H] + : C15H Eriodictyol [M + H] + : C15H Dihydrokaempferol [M + H] + : C15H129S Eriodictyol-7--sulphate [M + H] + : C15H129S Dihydrokaempferol-7--sulphate C15H129S Eriodictyol-3 --sulphate C15H129S Dihydrokaempferol- 4 --sulphate C21H C21H C21H Dehydroxylated taxifolin glucuronide 1 Dehydroxylated taxifolin glucuronide 2 Dihydrokaempferol 4 -glucuronide [M + H] + : [M+HCH-H] - : [M + H] + : [M + CH3HCH H] : MS 2 [ ]: (4.94), (98.68), (10), (59.71), (6.2), (4.94), (13.66); MS 3 [ ]: (10) MS 2 [ ]: (5.27), (7.48), (1.76), (10), (5.97), (0.75), (0.51) MS 2 [ ]: (4.63), (10), (4.57), (8.75), (12.96), (17.71); MS 3 [26672]: (39.08), (10), (15.31), (4.83), (60.18), (3.2) MS 2 [ ]: (41.18), (4.37), (10), (4.67) MS 2 [ ]: (10), (7.44), (95.06), (1.87), (2.27); MS 3 [ ]: (10), (5.36), (5.36), (7.99) MS 2 [ ]: (10), (3.42), (52.17); MS 3 [ ]: (45.3), (10) MS 2 [ ]: (5.6), (10), (5.6), (95.85), (5.6); MS 3 [ ]: (10), (44.81) [M + H] + : N [M + H] + : N [M + H] + : C21H Luteolin-7-- glucuronide [M + NH4] + : C21H Luteolin-3 /4 --glucuronide [M + NH4] + : C21H Luteolin-3 /4 --glucuronide [M + NH4] + : C22H Methyl luteolin glucuronide [M + NH4] + : MS 2 [ ]: (10.11), (17.49), (77.45), (10), (12.54) MS 2 [ ]: (17.38), (10), (1.78), (2.54), (1.52); MS 3 [ ]: (10) MS 2 [ ]: (14.77), (10), (1.75), (2.98), (5.34), (2.46), (4.04); MS 3 [ ]: (76.0), (10) MS 2 [ ]: (20.56), (10), (4.06), (5.89), (3.99), (1.34); MS 3 [ ]: (49.84), (49.84), (10), (49.84), (33.87) MS 2 [ ]: (30.71), (10), (9.33);MS 3 [ ]: (10), (35.16), (35.16)

11 S11 of S26 M11 7 M11 8 M11 9 M12 0 M12 1 M12 2 M12 3 M12 4 M12 5 M12 6 M12 7 M12 8 M12 9 M13 0 M13 1 tr(min) Formula Meas. m/z Identification ther Ions C15H Hydrogenated taxifolin [M + H] + : C16H Hydrogenated methyl taxifolin [M + H] + : C15H1410S Hydrogenated taxifolin sulphate C15H1410S Hydrogenated taxifolin sulphate 2 [M + H] + : C15H1410S Hydrogenated taxifolin sulphate 3 [M + H] + : C9H /4-Hydroxyphenylpropionic acid C9H /4-Hydroxyphenylpropionic acid C9H106S C9H106S C15H C15H Hydroxyphenylpropionic acid sulphate 3- Hydroxyphenylpropionic acid sulphate 3/4-Hydroxyphenylpropionic acid glucuronide 3/4-Hydroxyphenylpropionic acid glucuronide [M + HCH H] : [M + H] + : [M + HCH H] : [M + H] + : [M + CH3CH H] : [M + H] + : MS 2 [ ]: (27.3), (5.84), (84.98), (21.98), (19.64), (10.2), (10); MS 3 [ ]: (14.4), (57.2), (10) MS 2 [ ]: (2.11), (7.8), (13.17), (9.24), (13), (30.46), (10), (23.45) MS 2 [ ]: (10), (2.04), (14.45), (49.64), (8.54), (8.04); MS 3 [ ]: (10) MS 2 [ ]: (10), (34.27), (74.2), (5.95), (16.19), (18.96), (28.64); MS 3 [ ]: (13.22), (81.64), (14.5), (10), (5.33) MS 2 [ ]: (10), (3.84), (36.85), (19.37), (56.85), (19.71), (14.49), (20.95); MS 3 [ ]: (13.6), (13.6), (81.21), (13.6), (13.6), (13.6), (10) N MS 2 [ ]: (10), (5.32), (5.32) MS 2 [ ]: (10), (51.43), (2.54) MS 2 [ ]: (70.75), (10), (20.97); MS 3 [ ]: (10) [M + H] + : N [M+H] + : MS 2 [ ]: (39.84), (60.16), (39.84), (60.16), (60.16), (10) C9H86S p/m-coumaric acid sulphate [M + H] + : MS 2 [ ]: (2.83), (10), (42.98) C9H86S p/m-coumaric acid sulphate [M + H] + : MS 2 [ ]: (10), (17.51) C9H86S p/m-coumaric acid sulphate MS 2 [ ]: (10), (18.77); MS 3 [ ]: (10) C8H Dihydroxyphenylacetic acid [M + H] + : MS 2 [ ]: (10)

12 S12 of S26 M13 2 M13 3 M13 4 M13 5 M13 6 M13 7 M13 8 M13 9 M14 0 M14 1 M14 2 M14 3 M14 4 M14 5 M14 6 M14 7 M14 8 tr(min) Formula Meas. m/z Identification ther Ions C8H87S Dihydroxyphenylacetic acid sulfae C8H87S Dihydroxyphenylacetic acid sulfae C8H87S Dihydroxyphenylacetic acid sulfae C9H107S Homovanillic acid sulphate C9H Dihydrocaffeic acid [M + HCH H] - : [M + H] + : [M + HCH H] : [M + HCH H] : [M + H] + : [M + HCH H] : C9H107S Dihydrocaffeic acid sulphate 1 [M + H] + : C9H107S Dihydrocaffeic acid sulphate C11H13N Caffeic acid acetamide 1 [M + H] + : C11H13N Caffeic acid acetamide 2 [M + H] + : C11H13N Caffeic acid acetamide 3 [M + H] + : C9H C9H (3,4-Dihydroxyphenyl)-3- hydroxypropanoic acid 3-(3,4-Dihydroxyphenyl)-2- hydroxypropanoic acid C9H108S Caffeic acid hydrated sulphate C9H108S Caffeic acid hydrated sulphate C10H127S Dihydrogen ferulic acid sulphate MS 2 [ ]: (98.33), (3.62), (10), (2.41), (2.41) MS 2 [ ]: (40.61), (4.54), (10), (7.65) MS 2 [ ]: (25.11), (10), (4.22), (6.37) MS 2 [ ]: (68), (10), (69.26), (14.26), (0.79); MS 3 [ ]: (10), (28.58), (23.29) MS 2 [ ]: (10) MS 2 [ ]: (8.69), (77.44), (10), (2.90) MS 2 [ ]: (10), (55.09), (5.14); MS 3 [ ]: (10), (66.67) MS 2 [ ]: (10), (22.08), (22.08), (55.63), (33.33) MS 2 [ ]: (49.02), (49.02), (49.02), (61.76), (10) MS 2 [ ]: 22566(23.69), (94.08), (10), (35.54), (53.3), (23.69), (23.69) [M + H] + : MS 2 [ ]: (65.39), (10) [M + H] + : MS 2 [ ]: (10), (18.06) [M + HCH H] : [M + HCH H] : C10H128S Ferulic acid hydratedsulphate 1 [M + H] + : C10H128S Ferulic acid hydratedsulphate 2 MS 2 [ ]: (3.42), (29.18), (28.31), (10), (75.84), (2.75), (2.75), (46.29), (14.48), (2.75) MS 2 [ ]: (6.27), (41.39), (86.06), (10), (4.02), (78.31), (4.57), (2.29); MS 3 [ ]: (10) MS 2 [ ]: (10), (14.67), (1.61), (25.99), (12.43) MS 2 [ ]: (5.26), (5.26), (10), (92.66), (4.21), 15371(4.21), (5.26), (35.58), (4.21) MS 2 [ ]: (18.43), (36.68), (10), (27.64), (68.74)

13 S13 of S26 M14 9 M15 0 M15 1 M15 2 M15 3 M15 4 M15 5 M15 6 M15 7 M15 8 M15 9 M16 0 M16 1 M16 2 M16 3 M16 4 tr(min) Formula Meas. m/z Identification ther Ions C7H85S Hydroxybenzyl alcohol sulphate MS 2 [ ]: (10), (1.6), (3.98) C13H C13H C13H1611S C13H1611S C8H105S C8H105S Hydroxybenzyl alcohol glucuronide 1 Hydroxybenzyl alcohol glucuronide 2 Hydroxybenzyl alcohol glucuronide sulphate 1 Hydroxybenzyl alcohol glucuronide sulphate 2 Methyl hydroxybenzyl alcohol sulphate 1 Methyl hydroxybenzyl alcohol sulphate 2 [M + H] + : MS 2 [ ]: (21.37), (10), (21.37), (31.85), (89.52), (84.27) [M + H] + : MS 2 [ ]: (10), (53.08), (87.01) [M + Na] + : [M + Na] + : [M + HCH H] : [M + HCH H] : MS 2 [ ]: (13.22), (13.22), (26.43), (10), (13.22), (13.22) MS 2 [ ]: (3.3), (4.86), (10), (3.3), (3.3), (8.17) MS 2 [ ]: (9.57), (88.45), (10), (9.57) MS 2 [ ]: (8.38), (98.69), (10) C7H66S /4-Hydroxy benzoic acid sulphate MS 2 [ ]: (10) C7H66S /4-Hydroxy benzoic acid sulphate [M + 2H2 H] : MS 2 [ ]: (10) C8H87S Vanillic acid sulphate MS 2 [ ]: (10), (2.6), (2.6), (2.6), (2.6); MS 3 [ ]: (10) C8H87S Isovanillic acid sulphate MS 2 [ ]: (10), (11.02), (3.24) C31H C31H C31H C32H C31H2417S Dimer of taxiflolin and dehydroxylated methyl taxifolin Dimer of taxiflolin and methyl taxifolin 1 Dimer of taxiflolin and methyl taxifolin 2 Dimer of taxiflolin and dimethyl taxifolin Dimer of taxiflolin and methyl taxifolin sulphate 1 [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : MS 2 [ ]: (2.05), (6.63), (4.08), (36.21), (10), (33.47), (29.94), (25.92), (13.28), (28.46); MS 3 [ ]: (10) MS 2 [ ]: (15.33), (7.41), (10), (34.96), (6.04), (29.0); MS 3 [ ]: (26.67), (4), (26.67), (26.67), (26.67), (10) MS 2 [ ]: (16.72), (4.28), (10), (30.42), (8.15), (9.68), (25.51); MS 3 [ ]: (11.99), (21.7), (11.99), (10), (11.99) MS 2 [ ]: 33702(14.17), (15.62), (8.48), (10), (39.82), (11.27), (9.87), (35.53), (11.27) MS 2 [ ]: (19.07), (1.74), (5.56), (34.65), (3.82), (10), (28.94),

14 S14 of S26 M16 5 M16 6 M16 7 M16 8 M16 9 M17 0 M17 1 M17 2 M17 3 tr(min) Formula Meas. m/z Identification ther Ions C31H2417S C31H2417S C32H2617S C32H2617S C32H2617S C32H C32H C32H C32H Dimer of taxiflolin and methyl taxifolin sulphate 2 Dimer of taxiflolin and methyl taxifolin sulphate 3 Dimer of taxiflolin and dimethyl taxifolin sulphate 1 Dimer of taxiflolin and dimethyl taxifolin sulphate 2 Dimer of taxiflolin and dimethyl taxifolin sulphate 3 dehydroxylated methyl taxifolin 1 dehydroxylated methyl taxifolin 2 dehydroxylated methyl taxifolin 3 Dimer of methyl quercetin and methyl taxifolin 1 [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : (8.53), (23.27); MS 3 [ ]: (57.14), (71.43), (10) MS 2 [ ]: (55.56), (1.27), (44.86), (2.13), (10), (17.5), (2.57), (10.34); MS 3 [ ]: (10) MS 2 [ ]: (15.71), (5.18), (31.98), (11.98), (10), (31.91), (6.36), (19.51); MS 3 [ ]: (19.89), (20.25), (10), (17.46), (6.34), (5.92) MS 2 [ ]: (78.77), (1.13), (1.33), 33682(9.95), (55.43), (29.04), (34.88), (9.74), (7.27), (0.76), (15.99), (10), (6.5), (30.95), (4.0), (4.4), (7.26), (17.51) MS 2 [ ]: (10), (10.29), (38), (6.63), (69.79), (0.63), (8.55), (7.8); MS 3 [ ]: (15.18), (9.1), (6.03), (6.03), (10), (46.0), (6.03), (10.57), (24.22), (6.03) MS 2 [ ]: (54.36), 33683(5.99), (22.97), (9.76), (29.92), (10.78), (21.45), (10), (21.79), (8.23), (9.55); MS 3 [ ]: (41.76), (19.55), (28.29), (5.45), (10), (26.44), (7.71) MS 2 [ ]: (10), (58.45), (27.28), (94.74), (19.48), (15.59) MS 2 [ ]: 33665(13.11), (10), (34.37), (16.82), 30566(15.75), (71.1), (27.38), (15.72), (21.43); MS 3 [ ]: 27552(31.07), (10) MS 2 [ ]: (54.0), (4.69), (15.12), (10), (44.87), (5.82), (6.95), (34.84), (6.95), (6.95); MS 3 [ ]: (10), (66.67) MS 2 [ ]: (8.3), (10), (7.37), (5.51), (47.85), (15.64), (25.73), (5.55), (24.84), (3.72), (3.72), (7.44), (34.1), (7.37), (26.48), (3.72)

15 S15 of S26 M17 4 M17 5 M17 6 M17 7 M17 8 M17 9 M18 0 M18 1 M18 2 M18 3 M18 4 M18 5 tr(min) Formula Meas. m/z Identification ther Ions C32H C32H C32H C33H C33H C33H C32H C32H C33H C33H C33H C33H Dimer of methyl quercetin and methyl taxifolin 2 Dimer of methyl quercetin and methyl taxifolin 3 Dimer of methyl quercetin and methyl taxifolin 4 dehydroxylated dimethyl taxifolin 1 dehydroxylated dimethyl taxifolin 2 dehydroxylated dimethyl taxifolin 3 methyl taxifolin 1 methyl taxifolin 2 dimethyl quercetin 1 dimethyl quercetin 2 dimethyl taxifolin 1 dimethyl taxifolin 2 [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + 2H2 H] : [M+H] + : MS 2 [ ]: (17.71), (7.07), (10), (23.97), (2.99), (2.63), (7.08), (2.25), (2.25); MS 3 [ ]: (85.08), (57.46), (85.08), (85.08), (10) MS 2 [ ]: (5.04), (7.47), (10), (22.55), (21.29), (70.28), (25.02), (5.04), (7.47), (9.99) MS 2 [ ]: (6.78), (6.78), (13.64), (36.03), 30175(9.05), (10), (37.54), (18.23) MS 2 [ ]: (88.41), (33.48), (44.21), (10), (33.48) MS 2 [ ]: (5.27), (88.35), (42.47), (10), (10.34), (5.27), (30.98), (7.81), (10.34) MS 2 [ ]: (44.66), (29.31), (10), (16.73), (4.18), (31.4), (5.57), 28438(4.18), (4.89), (4.18) MS 2 [ ]: 33724(4.49), (72.09), (33.46), (2.45), (6.07), (10), (13.36), (46.07), (9.7), (4.05), (2.84), (7.3), (18.6);MS 3 [ ]: (10), (66.23) MS 2 [ ]: (12.42), (10.84), (9.56), (4.23), (10), (8.81), (12.72), (9.24), (13.0), (38.16), (10.66); MS 3 [ ]: (11.92), (10), (11.92) MS 2 [ ]: 33647(19.51), (99.94), (10), (58.75), (14.56), (20.67), 30288(12.19), (53.9), (12.64), (21.43), 2752(14.41); MS 3 [ ]: (10), (17.32), (34.63) MS 2 [ ]: (10), (88.75), (73.12), (62.58), (39.65), 30237(13.28), (37.93), (26.43), (18.26); MS 3 [ ]: (10) MS 2 [ ]: 3367(11.56), (10), (41.85), (16.57), (27.13), (11.9), (4.5), 27483(4.52), (4.03) MS 2 [ ]: (1.39), 33683(12.06), (10), (54.54), (31.1), (37.81), (18.72), 27536(9.99); MS 3 [ ]: 27485(10), (85.56)

16 S16 of S26 M18 6 M18 7 M18 8 M18 9 M19 0 M19 1 tr(min) Formula Meas. m/z Identification ther Ions C33H C33H C33H C33H C33H C32H2616S dimethyl taxifolin 3 dimethyl taxifolin 4 dimethyl taxifolin 5 dimethyl taxifolin 6 dimethyl taxifolin 7 dehydroxylated methyl taxifolin sulphate [M + H] + : [M + H] + : [M + H] + : [M + H] + : [M + H] + : MS 2 [ ]: 33676(14.44), (10), (33.72), (17.31), (39.46), (14.41), (4.34), (4.34), (4.34), (4.34) MS 2 [ ]: 33674(37.82), (10), (25.16), (14.76), (10.65), (23.06), (46.05), (8.55), (8.55), (14.76), (8.55) MS 2 [ ]: 33678(32.82), (10), (25.6), (22.99), (32.3), (14.74), (4.33), 27498(13.1); MS 3 [ ]: (24.67), (13.46), 27519(46.67), (44.51), (26.74), (13.46), (100), (99.63) MS 2 [ ]: 33688(8.27), (10), (32.11), (14.07), (21.35), (3.42), (4.38), (5.85), (11.67) MS 2 [ ]: (5.8), (10), (97.55), (3.82), (15.5), (14.51), 30155(3.82), (3.82) MS 2 [ ]: (10), (24.77), (10.6), (1.29), (22.41), (6.25); MS 3 [ ]: (10), (76.67), (15.27), (74.09), (28.07), (15.37) Abbreviations: N means that the fragment ions of a metabolite are not obtained, and its corresponding extracted ion chromatograms in negative ion mode is shown in Figure S4 S11.

17 S17 of S26 Table S2. Bioactivities of taxifolin metabolites. No. Identification B1 B2 B3 B4 B5 B6 B7 B8 B9 B10 B11 B12 M0 Taxifolin(parent compound) [1] [2,3] [4] [5,6] [7] [8] [2] [9] [10] [11] [12] [13,14] M1, M2 Taxifolin isomer [15] [16] [16,17] [18] [19] M70 Quercetin [1] [20] [21] [5] [22] [8,18] [23,24] [24] [25] [26] [27] [14,28] M73 Quercetin-4 --sulfate [29] M74 Quercetin-3 --sulfate [29] [30,31] [32] M75 Quercetin-3--sulfate [33] [34] [34] M76 Quercetin -glucuronide [29] [35] [30,31] [5] [32] [27] [36] M80 Isorhamnetin [37] [20] [21] [5] [8] [38] [39] [41] [14,28] [25,40] M83 Isorhamnetin-3--sulfate [29] M85 Isorhamnetin disulfate [33] [42] M104 Eriodictyol [1] [43] [44] [44,45] [46] [47] [23] [48] [49] [50] [51] M105 Dihydrokaempferol [52] [53] [53] [54,55] [56] M113 Luteolin-7--glucuronide [57] [58] M122 3/4-hydroxyphenylpropionic acid [59] [60,61] M123 3/4-hydroxyphenylpropionic acid [59] [60,61] M131 Dihydroxyphenylacetic acid [29] [62] [63] [59] [64] [65] M136 Dihydrocaffeic acid [29] [59] [60] [67] [62,66] Sum Abbreviations: B1, Antioxidant; B2, Anti-inflammatory; B3, Antitumor; B4, Cardioprotective; B5, Antidiabetic; B6, Antimicrobial; B7, Antiviral; B8, Hepatoprotective; B9, Prevention of Alzheimer disease; B10, Immunoregulatory; B11, Xanthine oxidase inhibitor; B12, Neuroprotective

18 S18 of S26 Table S3. The reported targets of taxifolin and its bioactive metabolites. Target Name Parent and Metabolites Nrf2 TAX [68], M80 [39], M70 [69], M104 [70] β-amyloid (Aβ) peptides TAX [71], M70 [71], M76 [72] PI3-K phosphoinositide 3-kinase (PI3K) TAX [73], M80 [74], M70 [75] MEK1 M70 [76], M80 [74] EGF receptor (EGFR) TAX [73], M70 [77] heme oxygenase-1 (H-1), M70 [78], M104 [70] microrna-155 M70 [79], M80 [79] TBK1-kinase M70 [80], M104 [80] Table S4. Targets of quercetin (M70) as predicted by PharmMapper. Rank PDB ID Fit Score Target Name 2 1uzf Angiotensin-converting enzyme 64 1j1b Glycogen synthase kinase-3 beta 139 1fcm Beta-lactamase 269 1c9c Beta-secretase m4h 2.85 Aspartate aminotransferase Event#: 4 MS(E-) Ret. Time: Scan#: 3972 Inten.(x10,000,000) m/z Event#: 5 MS/MS(E-) Ret. Time: Scan#: 3973 Precursor: Cutoff: Inten.(x1,000,000) (7) m/z Event#: 6 MS 3 (E-) Ret. Time: Scan#: 3974 Precursor: Cutoff: Inten m/z Figure. S1. Negative MS n spectra of taxifolin.

19 S19 of S26 Figure S2. Proposed fragmentation pathways of taxifolin in negative ion detection mode mass spectra.

20 S20 of S26 (A) Event#: 5 MS/MS(E-) Ret. Time: Scan#: 6221 Precursor: Cutoff: Inten m/z Event#: 6 MS3(E-) Ret. Time: Scan#: 6222 Precursor: Cutoff: Inten.(x10,000) (-) MS 2 of M160 (-) MS 3 of M m/z (B) H H H H H H H H Chemical Formula: C 15 H Exact Mass: H Chemical Formula: C 15 H Exact Mass: H Chemical Formula: C 14 H Exact Mass: CH 3 H H H CH 3 CH 3 CH 3 H H H H H H Chemical Formula: C 31 H Exact Mass: H H - Chemical Formula: C 16 H 11 7 Exact Mass: CH 3 H H - Chemical Formula: C 16 H 9 6 Exact Mass: Chemical Formula: C 13 H 7 4 Exact Mass: H - Chemical Formula: C 16 H 11 6 Exact Mass: Figure S3. Proposed fragmentation pathways of M160 in negative ion detection mode; (A) MS n spectra of M160 (B) fragmentation pathways, characteristic fragment ions, and proposed structure (1.00) 5.0 (A) (1.00) min M24 (B) min Figure S4. Extracted ion chromatograms at m/z in negative ion mode of urine samples: (A) blank group; (B) drug group.