Background
Methods
Information on experimental design and resources
Chemicals, materials and reagents
Preparation of GQD and FGQD extracts
Animal handing and biochemical parameters related to T2DM measurement
Statistical analysis
Preparation of standard and sample solutions for HPLC–MS and HPLC analysis
Untargeted metabolomics analysis by HPLC Q Exactive MS
Targeted quantification analysis by HPLC
Data processing and analysis
Results
Antidiabetic effect
Group | FINS (mIU/L) | ISI | HOMA-IR |
---|---|---|---|
NC | 4.92 ± 0.74 | − 3.38 ± 0.24 | 1.33 ± 0.30 |
DM | 9.88 ± 0.58** | − 5.24 ± 0.22** | 8.59 ± 1.75** |
HM | 7.17 ± 0.54## | − 4.47 ± 0.36## | 3.98 ± 1.07## |
GQD | 6.78 ± 0.35☆☆ | − 4.52 ± 0.23☆☆ | 4.18 ± 0.95☆☆ |
FGQD | 5.86 ± 0.55△△▲▲ | − 4.26 ± 0.18△△ | 3.20 ± 0.60△ |
Characterization of the chemical constituents in the GQD extract
Source |
t
R
(time) | Compound | Formula | Experimental
m/z
| Error ppm | Mode | MS/MS (m/z) | Structure type |
---|---|---|---|---|---|---|---|---|
P1 | 6.85 | 3’-Hydroxypuerarin-4’-O-glucoside | C27H30O15 | 595.16559 | − 0.263 | + | 475, 433, 415, 397, 379, 367, 337, 313, 283 | C-glycoside-O-glu |
P2 | 7.19 | Puerarin-4’-O-glucoside | C27H30O14 | 579.17120 | 0.636 | + | 417, 399, 381, 351, 321, 297, 267, 255 | C-glu-O-glu |
P3 | 7.80 | 3’-Methyoxy puerarin-4’-O-glucoside | C28H32O15 | 609.18237 | 1.598 | + | 447, 429, 411, 393, 381, 365, 351, 327, 297 | C-glycoside-O-glu |
P4 | 8.03 | Mirificin-4’-O-β-D-glucoside | C32H38O18 | 709.19830 | 0.859 | − | 457, 429 | C-glycoside-O-glu |
P5 | 8.33 | Daidzein-4’,7-O-glucoside | C27H30O14 | 579.17084 | 0.014 | + | 417, 255 | O-glu |
P6 | 8.77 | 3’-Hydroxypuerarin* | C21H20O10 | 433.11276 | − 0.377 | + | 415, 397, 379, 367, 337, 313, 283 | C-glu |
P7 | 8.89 | 3’-Methoxy-4’-O-glucoside-daidzin | C28H32O15 | 609.18262 | 2.008 | + | 447, 285 | O-glu |
P8 | 9.63 | 3’-Hydroxypuerarin xyloside | C26H28O14 | 565.15509 | − 0.163 | + | 433, 415, 397, 379, 367, 337, 313, 283 | C-glycoside-O-xyl |
C1 | 10.07 | Dihydro-11-Hydroxy-stepholidine-glucoside | C25H31NO10 | 506.20200 | − 0.073 | + | 344, 326, 295, 277 | alkaloid-O-glu |
P9 | 10.30 | 6’’-O-α-D-glucopyranosylpuerarin | C27H30O14 | 579.17059 | − 0.242 | + | 417, 399, 381, 351, 321, 297, 267 | C-glu-O-glu |
P10 | 10.43 | 3’-Hydroxydaidzin | C21H20O10 | 433.11389 | 0.967 | + | 271 | O-glu |
P11 | 10.62 | Puerarin* | C21H20O9 | 417.11786 | − 0.356 | + | 399, 381, 363, 351, 321, 297, 267, 255 | C-glu |
P12 | 10.73 | Mirificin | C26H28O13 | 549.16016 | − 0.107 | + | 417, 399, 363, 351, 321, 297, 267 | C-glycoside-O-api |
P13 | 11.16 | 3’-Methoxypuerarin | C22H22O10 | 447.12827 | − 0.678 | + | 429, 411, 381, 351, 327, 297 | C-glu |
P14 | 11.22 | 6’’-O-Xylosylpuerarin | C26H28O13 | 549.15997 | − 0.541 | + | 417, 399, 363, 351, 321, 297, 267 | C-glycoside-O-xyl |
C2 | 11.52 | Magnoflorine* | C20H23NO4 | 342.16996 | − 0.189 | + | 297, 265, 250, 237 | alkaloid |
P15 | 11.65 | 3’-Methoxypuerarin6’’-O-D-api | C27H30O14 | 579.17053 | − 0.521 | + | 447, 429, 411, 393, 381, 365, 351, 327, 297 | C-glu |
C3 | 12.03 | Norisocorydine | C19H22NO4 | 328.15411 | − 0.225 | + | 313, 298, 282 | alkaloid |
S1 | 12.11 | 2’,3,5,6’,7-Pentahydroxyflavanone | C15H12O7 | 303.05096 | 1.031 | − | 285, 275, 217, 177 | flavanones aglycone |
P16 | 12.15 | 3’-Methoxydaidzin 6’’-O-D-api | C27H34O11 | 579.17108 | 0.428 | + | 255 | O-glu |
P17 | 12.19 | 5’-Hydroxypuerarin | C21H20O10 | 433.11328 | 0.824 | + | 415, 397, 367, 313, 283 | C-glu |
P18 | 12.57 | Daidzin* | C21H20O9 | 417.11807 | 0.147 | + |
255
| O-glu |
C4 | 12.75 | 11-Hydroxy-stepholidine-glucoside | C25H30NO10 | 505.18610 | − 0.322 | + | 342, 324, 275 | alkaloid-O-glu |
P19 | 13.02 | Genistein-8-C-xyl-glucoside | C26H28O14 | 565.15619 | 1.008 | + | 433, 415, 397, 367, 313, 283 | C-glycoside O-xyl |
P20 | 13.40 | BiochaninA-7-O-glucoside | C22H22O10 | 447.12933 | 1.692 | + | 285, 270, 253, 225 | O-glu |
C5 | 13.61 | O,O’-Dimethoxyl magnoflorine | C21H25NO5 | 372.18045 | − 0.099 | + | alkaloid | |
P21 | 13.72 | Genistein-8-C-api-glucoside | C26H28O14 | 565.15503 | − 0.269 | + | 433, 415, 367, 337, 313, 283 | C-glycoside-O-api |
P22 | 14.48 | PuerosideA | C29H34O14 | 607.20148 | − 0.652 | + | 592, 461, 299, 281, 253 | O-glu |
P23 | 14.53 | Daidzein 4’-O-glucoside | C21H20O9 | 417.11771 | − 0.229 | + | 255, 199 | O-glu |
G1 | 14.79 | Liquiritengin-glucopyranoside-(1→2)-β-D apiofuranoside | C26H30O13 | 549.16101 | 1.352 | − | 255, 153, 135, 119 | O-glu-O-api |
S2 | 14.96 | Chrysin-6-C-pen-8-C-hex | C26H28O13 | 547.14851 | 2.180 | − | 487, 457, 427, 367, 337, 281 | C-glu |
S3 | 15.00 | Viscidulin I | C15H10O7 | 301.03522 | 1.241 | − | 283, 273, 257, 229, 193, 151 | flavone aglycone |
C6 | 15.10 | 13-Hydroxyepiberberine | C20H17NO5 | 352.11787 | − 0.149 | + | 336, 322, 294 | alkaloid |
S4 | 15.36 | Chrysin-6-C-pen-8-C-hex | C26H28O13 | 547.14600 | 2.527 | − | 487, 457, 427, 367, 337, 281 | C-glu |
G2 | 15.38 | Liquiritin apioside | C26H30O13 | 549.16046 | 0.351 | − | 255, 153, 135, 119, 91 | O-glu |
G3 | 15.44 | Liquiritin* | C21H22O9 | 417.11917 | 2.784 | − | 255, 153, 135, 119 | O-glu |
S5 | 15.83 | Chrysin 6-C-α-L-arabinoside-8-C-β-D-glucoside | C26H28O13 | 547.14581 | 2.18 | − | 487, 457, 427, 367, 337, 281 | C-glu |
P24 | 15.90 | Neopuerarin | C21H20O9 | 417.11755 | − 1.100 | + | 399, 381, 363, 351, 321, 297, 267 | C-glu |
P25 | 15.97 | 6-D-xylose-Genistin | C26H28O14 | 565.15768 | 2.408 | + | 433, 271 | O-glu |
S6 | 16.14 | Scutellarein 7-β-D-glucuronoside | C21H18O12 | 461.07266 | 2.619 | − | 285, 267 | O-gluA |
S7 | 16.16 | Chrysin 6-C-β-L-arabinoside-8-C-β-D-glucoside | C26H28O13 | 547.14587 | 2.290 | − | 487, 457, 427, 367, 337, 281 | C-glu |
C7 | 16.28 | Stecepharine | C21H25NO5 | 372.18015 | − 0.399 | + | 222, 207, 189 | alkaloid |
S8 | 16.31 | Viscidulin III 2’-O-glucoside | C23H24O13 | 507.11469 | 2.707 | − | 345, 330, 315 | O-glu |
S9 | 16.43 | Acteoside | C29H36O15 | 623.19861 | 2.509 | − | 461, 161, 179 | O-glu |
P26 | 16.44 | Genistin* | C21H20O10 | 433.11334 | 0.417 | + |
271
| O-glu |
P27 | 16.46 | Kuzubutenolide A | C23H24O10 | 461.14017 | − 4.053 | + | 299, 281, 253, 239 | O-glu |
S10 | 16.52 | Chrysin 6-C-β-D-glucoside- 8-C-α-L-arabinoside | C26H28O13 | 547.14569 | 1.961 | − | 457, 427, 367, 337, 321 | C-glu |
P28 | 16.74 | Formononetin-8-C-glucoside-O-api | C27H30O13 | 563.17694 | 1.023 | + | 431, 413, 311, 281 | C-glu-O-api |
C8 | 16.88 | Groenlandicine | C19H15NO4 | 322.10712 | − 0.821 | + | 307, 279 | alkaloid |
C9 | 16.99 | Demethyleneberberine | C19H17NO4 | 324.12283 | -0.205 | + | 308, 266, 281 | alkaloid |
S11 | 17.02 | chrysin6-hexosyl-8-C-pentosyl | C26H28O13 | 547.14612 | 1.503 | − | 457, 427, 367, 337, 321 | C-glu |
P29 | 17.05 | Formononetin-8-C-glucoside-O-xyloside | C27H30O13 | 563.17554 | − 0.670 | + | 431, 413, 311, 281 | C-glu |
P30 | 17.10 | 6′′-O-Malonyl daidzin | C24H22O12 | 503.11804 | − 0.362 | + |
255
| O-glu |
S12 | 17.19 | Chrysin 6-C-β-D-glucoside-8-C-β-L- arabinoside | C26H28O13 | 547.14575 | 2.070 | − | 457, 427, 367, 337, 321 | C-glu |
P31 | 17.22 | 4′-Methoxypuerarin | C22H22O9 | 431.13364 | − 0.043 | + | 413, 395, 377, 335, 311, 281 | C-glu |
S13 | 17.34 | 5,2′,6′-Trihydroxy-7,8-dimmethoxy flavone -2′-glucoside | C23H24O12 | 491.11948 | 2.194 | − | 329, 314, 299 | O-glu |
S14 | 17.36 | Isoacteoside | C29H36O15 | 623.19843 | 1.383 | − | 461, 161, 179 | O-glu |
C10 | 17.38 | Oxyberberine | C20H17NO5 | 352.11789 | − 0.059 | + | 337, 336, 322, 308, 294 | alkaloid |
C11 | 17.64 | Oxidated palmatine | C21H21NO5 | 368.14893 | − 0.314 | + | 352, 336 | alkaloid |
G4 | 18.06 | Pyrroside B | C26H30O14 | 565.15649 | 2.315 | − | 271, 151 | O-glu |
G5 | 18.59 | 5-Hydroxylliquiritin | C21H22O10 | 433.11404 | 1.117 | − | 271, 151, 119 | O-glu |
S15 | 19.12 | 5,7,2′,6′-Tetrahydroxyflavone | C15H10O6 | 285.04050 | 3.984 | − | 241, 199, 133, 151 | flavone aglycone |
P32 | 19.21 | 6′′-O-Acetyl daidzin | C23H22O10 | 459.12839 | − 0.183 | + |
255
| O-glu |
C12 | 19.49 | Columbamine | C20H19NO4 | 338.13849 | − 0.576 | + | 323, 308, 294 | alkaloid |
S16 | 19.58 | 5,7,2′-Trihydroxy-6-methoxyflavone 7-O-glucuronide | C22H20O12 | 475.08975 | 0.848 | − | 299, 284, 175, 113 | O-gluA |
C13 | 19.70 | Epiberberine | C20H18NO4 | 336.12274 | − 0.876 | + | 321, 320, 292 | alkaloid |
C14 | 19.98 | Coptisine* | C19H13NO4 | 320.09174 | 0.017 | + | 292, 262 | alkaloid |
C15 | 20.13 | Jatrorrhizine | C20H19NO4 | 338.13855 | − 0.398 | + | 323, 308, 294 | alkaloid |
P33 | 20.81 | Sophoraside A or isomer | C24H26O10 | 473.14496 | 0.737 | − | 311, 267, 252 | O-glu |
G6 | 20.89 | Isoliquiritin apioside | C26H30O13 | 549.16150 | 0.191 | − | 255, 153, 135, 119, 91 | O-glu |
S17 | 21.68 | Scutellarein* | C15H10O6 | 285.04071 | 4.720 | − | 267, 239, 166, 137, 117 | flavone aglycone |
P34 | 22.04 | Ononin* | C22H22O9 | 431.13361 | − 0.101 | + |
269
| O-glu |
G7 | 22.13 | Licuraside | C26H30O13 | 549.16040 | 0.133 | − | 255, 153, 135, 119 | O-glu |
G8 | 22.23 | Isoliquiritin* | C21H22O9 | 417.11948 | 1.471 | − | 255, 153, 135, 119 | O-glu |
S18 | 22.44 | Baicalein 7-β-D-glucoside | C21H20O10 | 433.11276 | − 0.163 | + |
271
| O-glu |
S19 | 22.50 | Baicalin* | C21H18O11 | 445.09216 | 0.028 | − | 269, 241, 223, 175, 113 | O-gluA |
S20 | 22.60 | Eriodictyol | C15H12O6 | 287.05630 | 1.285 | − | 218, 161, 125 | flavanones aglycone |
C16 | 22.65 | Worenine+CH2+2H | C21H21NO4 | 352.15433 | − 0.285 | + | 334, 320 | alkaloid |
P35 | 23.45 | Daidzein* | C15H10O4 | 255.06509 | − 0.374 | + | 227, 199, 181, 153 | flavanones aglycone |
G9 | 23.47 | Neoisoliquiritin | C21H22O9 | 417.11954 | 1.531 | − | 255, 153, 119 | O-glu |
C17 | 23.56 | Worenine | C20H15NO4 | 334.10721 | − 0.174 | + | 319, 306, 291 | alkaloid |
G10 | 23.59 | Licochalcone B | C16H14O5 | 285.07687 | 3.929 | − | 270, 253, 191, 150 | chalcones aglycone |
G11 | 24.13 | Licorice glycosideB | C35H36O15 | 695.19727 | 0.223 | − | 549, 531, 399, 255 | O-glu |
G12 | 24.18 | Liquiritigenin* | C15H11O4 | 255.06641 | 4.801 | − | 237, 153, 135, 119, 91 | flavanones aglycone |
P36 | 24.21 | Isoononin | C22H22O9 | 431.13461 | 0.951 | + |
269
| O-glu |
C18 | 24.53 | Palmatine* | C21H21NO4 | 352.15417 | − 0.468 | + | 337, 336, 322, 308, 294 | alkaloid |
P37 | 24.69 | BiochaninA | C16H12O5 | 283.06110 | 3.533 | − | 268, 240, 211 | flavone aglycone |
P38 | 24.82 | Apigenin* | C15H10O5 | 269.04568 | 1.230 | − | 241, 225, 213, 197 | flavone aglycone |
S21 | 24.88 | Naringenin 7-O-β-D-glucuronide | C21H20O11 | 447.09454 | 2.352 | − | 271, 243, 113 | O-gluA |
C19 | 24.96 | Berberine* | C20H17NO4 | 336.12274 | − 0.876 | + | 321, 320, 306, 292 | alkaloid |
C20 | 25.27 | Demethylcoptichine | C30H25NO8 | 528.16552 | − 0.073 | + | 334, 319, 304 | alkaloid |
S22 | 25.52 | Norwogonin-8-Oglucuronide | C21H18O11 | 445.07779 | 2.814 | − | 269, 251, 241 | O-gluA |
S23 | 26.15 | Trihydroxymethoxyflavone-Oglucoside | C22H22O11 | 461.10977 | 1.932 | − | 299, 284, 283, 211, 173 | O-glu |
S24 | 26.18 | Hydroxyl oroxylin A-7-O-glucuronide | C22H20O12 | 475.08813 | 1.028 | − | 299, 284 | O-gluA |
S25 | 26.40 | 4′-Hydroxylwogonin | C16H12O6 | 299.05606 | 1.0455 | − | 271, 227, 211, 165, 133 | O-gluA |
S26 | 26.57 | Norwogonin-7-Oglucuronide | C21H18O11 | 445.07748 | 2.117 | − | 269, 251, 241 | O-gluA |
S27 | 26.98 | Chrysin-7-O-glucuronide | C21H18O10 | 431.09763 | 0.828 | + |
255
| O-gluA |
S28 | 27.00 | Oroxylin A-7-O-glucuronide | C22H20O11 | 459.09357 | 3.001 | − | 283, 268, 175, 113, 85 | O-gluA |
S29 | 27.38 | Hydroxyl wogonoside | C22H20O12 | 475.08810 | 0.998 | − | 299, 284 | O-gluA |
P39 | 27.49 | Isoformononetin | C16H12O4 | 267.06650 | 1.315 | − | 252, 223, 199 | isoflavone aglycone |
C21 | 27.84 | 13-Methylberberine | C21H19NO4 | 350.13864 | − 0.045 | + | 335, 334, 320, 318, 306 | alkaloid |
S30 | 28.09 | Baicalein 6-O-glucuronide | C21H18O11 | 445.07791 | 3.083 | − | 269, 241, 225, 197 | O-gluA |
C22 | 28.16 | Demethylcoptichine | C30H25NO8 | 528.16595 | 1.244 | + | 334, 319, 304 | alkaloid |
S31 | 28.52 | Wogonoside* | C22H20O11 | 459.09348 | 2.815 | − | 283, 268, 175, 113, 85 | O-gluA |
S32 | 29.11 | 5,7-Dihydroxy-6,8-dimethoxyflavone-7-O glucuronide | C23H22O12 | 489.10565 | 2.898 | − | 313, 298, 283 | O-gluA |
P40 | 30.47 | Genistein* | C15H10O5 | 269.04572 | 4.721 | − | 241, 225, 183, 159 | isoflavone aglycone |
S33 | 30.53 | 5,7,4′-Trihydroxy-8-methoxyflavone | C16H12O6 | 299.05637 | 4.532 | − | 284, 231, 136, 94 | flavone aglycone |
G13 | 31.48 | Licorice saponin A3 | C48H72O21 | 983.44867 | 0.435 | − | 821, 351 | O-gluA-gluA |
S34 | 31.88 | Norwogonin | C15H10O5 | 269.04578 | 1.330 | − | 251, 241, 223 | flavone aglycone |
G14 | 32.14 | 22β-Acetoxylglycyrrhizic acid | C44H64O18 | 879.40295 | 2.341 | − |
351
| O-gluA-gluA |
S35 | 32.20 | 5,7,2′-Trihydroxy-6-methoxyflavone | C16H12O6 | 299.05630 | 1.285 | − | 284, 255 | flavone aglycone |
G15 | 32.19 | Licorice saponin G2 | C42H62O17 | 837.39185 | 1.819 | − | 351, 193 | O-gluA-gluA |
S36 | 32.19 | Trihydroxydimethoxyflavone | C17H14O7 | 329.06674 | 3.528 | − | 314, 299 | flavone aglycone |
S37 | 32.40 | Baicalein* | C15H10O5 | 269.06000 | − 0.100 | − | 251, 241, 223, 213, 197 | flavone aglycone |
S38 | 32.64 | Trihydroxy-methoxyflavone | C16H12O6 | 299.05627 | 4.198 | − | 284, 165, 137 | flavone aglycone |
G16 | 32.73 | Isoliquiritigenin* | C15H11O4 | 255.06633 | 4.566 | − | 237, 153, 119, 91 | chalcones aglycone |
G17 | 32.80 | Glycyrrhizic acid* | C42H62O16 | 821.39655 | 1.385 | − |
351
| O-gluA-gluA |
P41 | 32.97 | Formononetin* | C16H12O4 | 267.06647 | 4.810 | − | 252, 223 | isoflavone aglycone |
G18 | 33.07 | Glycyrrhizin isomer | C42H62O16 | 821.39642 | 1.227 | − |
351
| O-gluA-gluA |
G19 | 33.39 | Licorice saponin C2 | C42H62O15 | 805.40094 | 0.433 | − | 351, 193 | O-gluA-gluA |
G20 | 33.50 | Licorice saponin B2 | C42H64O15 | 807.41724 | 1.093 | − | 351, 193 | O-gluA-gluA |
S39 | 33.54 | Skullcapflavone | C18H16O7 | 343.08255 | 1.321 | − | 328, 313, 298, 285 | flavone aglycone |
G21 | 33.72 | Liconeolignan | C21H22O5 | 353.13947 | 1.120 | − | 338, 321, 295, 283, 269 | others |
S40 | 33.77 | Wogonin* | C16H12O5 | 283.06140 | 4.593 | − | 268, 239, 163 | flavone aglycone |
S41 | 33.78 | Chrysin | C15H10O4 | 255.06497 | − 0.215 | + | 238, 214 | flavone aglycone |
S42 | 33.90 | Dihydroxy-dimethoxyflavone | C17H14O6 | 315.08600 | − 0.315 | + | 300, 285 | flavone aglycone |
C23 | 33.95 | Berberastine | C20H18NO5 | 352.11774 | − 0.209 | + | 336, 322, 308 | alkaloid |
S43 | 34.03 | Oroxylin A | C16H12O5 | 283.06137 | 4.487 | − | 268, 239, 163 | flavone aglycone |
S44 | 34.21 | Tenaxin I | C18H16O7 | 343.08252 | 1.321 | − | 268, 239, 163 | flavone aglycone |
G22 | 34.40 | Licoisoflavone A | C20H18O6 | 353.10330 | 1.335 | − | 284, 267, 243, 216, 201, 83 | isoflavone aglycone |
G23 | 34.45 | Licochalcone A | C21H22O4 | 337.14484 | 1.404 | − | 305, 281, 243, 229, 201 | chalcones aglycone |
G24 | 35.03 | Glabrone | C20H16O5 | 335.09271 | 1.310 | − | 305, 291, 275, 213, 199, 107 | isoflavone aglycone |
G25 | 35.03 | Licoisoflavone B | C20H16O6 | 351.08755 | 1.235 | − | 283, 265, 241, 199, 83 | isoflavone aglycone |
Isoflavone glycosides
Flavone glycosides
Free flavones
Alkaloids
Triterpene saponins
Others
Multivariate statistical analysis
Targeted quantification analysis
Compound | 0 h | 48 h |
---|---|---|
Puerarin | 20.30 ± 0.05 | 23.57 ± 0.02* |
Daidzin | 3.67 ± 0.08 | n.d.*** |
Daidzein | 0.50 ± 0.02 | 3.80 ± 0.01*** |
Liquiritin | 0.80 ± 0.06 | 0.48 ± 0.02* |
Liquiritigenin | 0.17 ± 0.05 | 0.50 ± 0.01* |
Coptisine | 2.23 ± 0.12 | 2.68 ± 0.003 |
Palmatine | 2.01 ± 0.03 | 2.36 ± 0.15 |
Berberine | 7.70 ± 0.03 | 8.10 ± 0.02 |
Baicalin | 10.80 ± 0.02 | 11.85 ± 0.01 |
Baicalein | 1.26 ± 0.04 | 1.27 ± 0.03 |