3.4. Identification of Flavonoids
Flavonoids were identified via UV-vis spectral survey, HR-MS, LC-MS, acid hydrolysis, NMR and/or HPLC and TLC comparisons with authentic samples. NMR spectra and signal assignment for flavonoids are shown in
Table 1 and
Supplementary Materials Figures S1–S5. The origins of the authentic samples used in this survey were as follows: kaempferol from
Dianthus caryophyllus flowers (as hydrolysate) [
22], kaempferol 3-
O-glucoside and quercetin 3-
O-glucoside from
Cyrtomium spp. fronds [
23], kaempferol 3,7-di-
O-rhamnoside from
Hylotelephium sieboldii leaves and stems [
24], kaempferol 3-
O-glucoside-7-
O-rhamnoside from
Lathyrus japonicus leaves [
25], quercetin and herbacetin from Extrasynthese (France), myricetin 3-
O-glucoside from
Corylopsis spp. leaves [
26], and cyanidin 3-
O-glucoside from
Acer spp. leaves [
27].
3.4.1. Herbacetin 3-O-glucoside-8-O-arabinoside (1)
TLC (Rf): 0.31 (BAW), 0.55 (BEW), 0.56 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark greenish yellow. HPLC (retention times, tR): 6.72 min (solv. I). UV: λmax (nm) MeOH 272, 357; +NaOMe 281, 326, 407 (inc.); +AlCl3 280, 310, 354, 407; +AlCl3/HCl 280, 308, 347, 406; +NaOAc 280, 316, 398; +NaOAc/H3BO3 274, 364. HR-MS (ESI) [M − H]− calcd. for C26H27O16: 595.1299, Found: 595.1291. LC-MS: m/z 597 [M + H]+, 595 [M − H]−, m/z 435 [M-162 + H]+ and m/z 303 [M-162-132 + H]+. 1H NMR (800MHz, DMSO-d6): δ 8.23 (2H, d, J = 8.8 Hz, H-2′,6′), 6.85 (2H, d, J = 8.8 Hz, H-3′,5′), 6.02 (1H, s, H-6), 5.41 (1H, d, J = 7.2 Hz, glucosyl H-1), 4.58 (1H, d, J = 6.4 Hz, arabinosyl H-1), 3.81 (1H, dd, J = 4.0 and 12.0 Hz, arabinosyl H-5b), 3.68 (1H, brd, J = 8.0 Hz, arabinosyl H-2), 3.67 (1H, brd, J = 6.4 Hz, arabinosyl H-4), 3.58 (1H, brd, J = 10.4 Hz, glucosyl H-6b), 3.48 (1H, brd, J = 10.4 Hz, arabinosyl H-3), 3.47 (1H, brd, J = 10.4 Hz, arabinosyl H-5a), 3.37 (1H, dd, J = 4.8 and 8.7 Hz, glucosyl H-6a), 3.22 (1H, m, glucosyl H-3), 3.21 (1H, m, glucosyl H-2), 3.11 (1H, m, glucosyl H-5), 3.10 (1H, m, glucosyl H-4). 13C NMR (200 MHz, DMSO-d6): (herbacetin) δ 154.8 (C-2), 132.9 (C-3), 176.5 (C-4), 164.8 (C-5), 100.9 (C-6), 156.8 (C-7), 125.5 (C-8), 148.5 (C-9), 100.5 (C-10), 121.2 (C-1′), 131.0 (C-2′), 115.0 (C-3′), 159.9 (C-4′), 115.0 (C-5′), 131.0 (C-6′); (3-O-glucose) δ 101.6 (C-1), 74.3 (C-2), 76.6 (C-3), 69.9 (C-4), 77.5 (C-5), 60.9 (C-6); (8-O-arabinose) δ 106.3 (C-1), 70.7 (C-2), 72.3 (C-3), 67.0 (C-4), 65.5 (C-5).
3.4.2. Herbacetin 3-O-glucoside-8-O-xyloside (2)
TLC (Rf): 0.47 (BAW), 0.63 (BEW), 0.55 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 greenish yellow. HPLC (tR): 6.46 min (solv. I). UV: λmax (nm) MeOH 271, 351; +NaOMe 281, 325, 407 (inc.); +AlCl3 274, 308, 351, 405sh; +AlCl3/HCl 275, 310, 350, 404sh; +NaOAc 280, 308, 384; +NaOAc/H3BO3 273, 368. HR-MS (ESI) [M − H]− calcd. for C26H27O16: 595.1299, Found: 595.1278. LC-MS: m/z 597 [M + H]+, 595 [M − H]−, m/z 463 [M-132-H]−, m/z 435 [M-162 + H]+ and m/z 303 [M-162-132 + H]+. 1H NMR (800MHz, DMSO-d6): δ 8.24 (2H, d, J = 8.9 Hz, H-2′,6′), 6.82 (2H, d, J = 8.9 Hz, H-3′,5′), 5.75 (1H, s, H-6), 5.35 (1H, d, J = 7.6 Hz, glucosyl H-1), 4.34 (1H, d, J = 7.3 Hz, xylosyl H-1), 3.77 (1H, dd, J = 5.6 and 11.2 Hz, xylosyl H-5b), 3.58 (1H, brd, J = 11.2 Hz, glucosyl H-6b), 3.37 (1H, m, glucosyl H-6a), 3.34 (1H, m, xylosyl H-4), 3.19 (1H, m, glucosyl H-3), 3.18 (1H, m, xylosyl H-2), 3.18 (1H, m, glucosyl H-2), 3.18 (1H, m, xylosyl H-3), 3.10 (1H, m, glucosyl H-5), 3.10 (1H, m, xylosyl H-5a), 3.09 (1H, m, glucosyl H-4). 13C NMR (200 MHz, DMSO-d6): (herbacetin) δ 153.4 (C-2), 132.6 (C-3), 175.7 (C-4), 164.0 (C-5), 101.7 (C-6), 157.1 (C-7), 127.2 (C-8), 148.3 (C-9), 101.7 (C-10), 121.6 (C-1′), 130.8 (C-2′), 114.8 (C-3′), 159.4 (C-4′), 114.8 (C-5′), 130.8 (C-6′); (3-O-glucose) δ 102.1 (C-1), 74.3 (C-2), 76.7 (C-3), 70.0 (C-4), 77.4 (C-5), 60.9 (C-6); (8-O-xylose) δ 108.4 (C-1), 73.8 (C-2), 76.6 (C-3), 69.2 (C-4), 66.3 (C-5).
3.4.3. Herbacetin 3-O-xyloside-8-O-glucoside (3)
TLC (Rf): 0.44 (BAW), 0.57 (BEW), 0.63 (15%HOAc); color UV (365 nm) dark purple, UV/NH
3 dark greenish yellow. HPLC (
tR): 7.48 min (solv. I). UV: λmax (nm) MeOH 271, 355; +NaOMe 281, 327, 404 (inc.); +AlCl
3 279, 309, 353, 407; +AlCl
3/HCl 279, 308, 348, 404; +NaOAc 280, 320, 400; +NaOAc/H
3BO
3 274, 366. HR-MS (ESI) [M − H]
− calcd. for C
26H
27O
16: 595.1299, Found: 595.1290. LC-MS:
m/
z 597 [M + H]
+, 595 [M − H]
−,
m/
z 435 [M-162 + H]
+ and
m/
z 303 [M-162-132 + H]
+.
1H and
13C NMR, see
Table 1. Aglycone of
3 (herbacetin). HPLC (
tR): 6.75 min (solv. II). UV: λmax (nm) MeOH 248, 275, 299, 370; +NaOMe decomp.; +AlCl
3 260sh, 341, 378sh, 433; +AlCl
3/HCl 249, 268sh, 306, 365, 429; +NaOAc 274sh, 304, 372; +NaOAc/H
3BO
3 310, 381.
3.4.4. Herbacetin 3-O-glucoside-8-O-(2‴-acetylxyloside) (4)
TLC (Rf): 0.59 (BAW), 0.71 (BEW), 0.56 (15%HOAc); color UV (365 nm) dark purple, UV/NH
3 dark greenish yellow. HPLC (
tR): 15.49 min (solv. I). UV: λmax (nm) MeOH 272, 354; +NaOMe 282, 326, 408 (inc.); +AlCl
3 274, 309, 354, 406sh; +AlCl
3/HCl 276, 308, 350, 405sh; +NaOAc 280, 309, 385; +NaOAc/H
3BO
3 274, 319, 364. HR-MS (ESI) [M − H]
− calcd. for C
28H
29O
17: 637.1405, Found: 637.1380. LC-MS:
m/
z 639 [M + H]
+, 637 [M − H]
−,
m/
z 477 [M-162 + H]
+ and
m/
z 303 [M-42-132-162 + H]
+.
1H and
13C NMR, see
Table 1. Aglycone of
4 (herbacetin). HPLC (
tR): 6.75 min (solv. II). UV: λmax (nm) MeOH 249, 276, 301, 374; +NaOMe decomp.; +AlCl
3 267sh, 339, 374sh, 452; +AlCl
3/HCl 268, 306sh, 359, 436; +NaOAc 273sh, 323, 374; +NaOAc/H
3BO
3 310, 377.
3.4.5. Gossypetin 3-O-glucoside-8-O-xyloside (5)
TLC (Rf): 0.28 (BAW), 0.49 (BEW), 0.48 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark greenish yellow. HPLC (tR): 5.34 min (solv. I). UV: λmax (nm) MeOH 260, 268sh, 364; +NaOMe 283, 418 (inc.); +AlCl3 278, 440; +AlCl3/HCl 273, 304sh, 363, 410; +NaOAc 279, 328, 408; +NaOAc/H3BO3 268, 388. LC-MS: m/z 613 [M + H]+, 611 [M − H]−, m/z 479 [M-132-H]−, m/z 451 [M-162 + H]+ and m/z 319 [M-162-132 + H]+. 1H NMR (800 MHz, DMSO-d6): δ 7.84 (1H, d, J = 2.4 Hz, H-2′), 7.75 (1H, dd, J = 2.4 and 8.8 Hz, H-6′), 6.80 (1H, d, J = 8.0 Hz, H-5′), 5.80 (1H, s, H-6), 5.38 (1H, d, J = 8.0 Hz, glucosyl H-1), 4.42 (1H, d, J = 8.0 Hz, xylosyl H-1), 3.83 (1H, dd, J = 4.8 and 11.2 Hz, xylosyl H-5b), 3.60 (1H, brd, J = 9.6 Hz, glucosyl H-6b), 3.38 (1H, m, xylosyl H-4), 3.37 (1H, t, J = 5.6 Hz, glucosyl H-6a), 3.28 (1H, t, J = 8.0 Hz, glucosyl H-3), 3.25 (1H, t, J = 8.0 Hz, xylosyl H-2), 3.23 (1H, t, J = 8.8 Hz, glucosyl H-2), 3.16 (1H, t, J = 8.8 Hz, xylosyl H-3), 3.12 (1H, m, glucosyl H-5), 3.11 (1H, m, xylosyl H-5a), 3.11 (1H, m, glucosyl H-4). 13C NMR (200 MHz, DMSO-d6): (gossypetin) δ 154.4 (C-2), 132.8 (C-3), 176.1 (C-4), 157.0 (C-5), 101.0 (C-6), 157.0 (C-7), 126.5 (C-8), 148.4 (C-9), 99.8 (C-10), 121.6 (C-1′), 117.2 (C-2′), 144.6 (C-3′), 148.2 (C-4′), 114.9 (C-5′), 121.8 (C-6′); (3-O-glucose) δ 101.8 (C-1), 74.1 (C-2), 76.6 (C-3), 69.9 (C-4), 77.4 (C-5), 61.0 (C-6); (8-O-xylose) δ 107.9 (C-1), 73.8 (C-2), 76.5 (C-3), 69.2 (C-4), 66.2 (C-5).
3.4.6. Gossypetin 3-O-glucoside-8-O-arabinoside (6)
TLC (Rf): 0.21 (BAW), 0.44 (BEW), 0.46 (15%HOAc); color UV (365 nm) dark purple, UV/NH
3 dark greenish yellow. HPLC (
tR): 5.74 min (solv. I). UV: λmax (nm) MeOH 262, 271, 365; +NaOMe 282, 420 (inc.); +AlCl
3 274, 304sh, 370, 414sh; +AlCl
3/HCl 273, 305sh, 364, 407sh; +NaOAc 280, 327, 393; +NaOAc/H
3BO
3 268, 390. HR-MS (ESI) [M − H]
− calcd. for C
26H
27O
17: 611.1248, Found: 611.1227. LC-MS:
m/
z 613 [M + H]
+, 611 [M − H]
−,
m/
z 479 [M-132-H]
−,
m/
z 451 [M-162 + H]
+ and
m/
z 319 [M-162-132 + H]
+.
1H and
13C NMR, see
Table 1. Aglycone of
6 (gossypetin). HPLC (
tR): 4.99 min (solv. II). UV: λmax (nm) MeOH 259, 280, 298, 341, 377; +NaOMe decomp.; +AlCl
3 287, 336, 388, 466; +AlCl
3/HCl 273, 286sh, 304sh, 371, 443; +NaOAc 287, 375; +NaOAc/H
3BO
3 310, 361.
3.4.7. Gossypetin 3-O-glucoside-8-O-(2‴-acetylxyloside) (7)
TLC (Rf): 0.51 (BAW), 0.63 (BEW), 0.64 (15%HOAc); color UV (365 nm) dark purple, UV/NH
3 dark greenish yellow. HPLC (
tR): 10.13 min (solv. I). UV: λmax (nm) MeOH 261, 267sh, 360; +NaOMe 283, 325sh, 421 (inc.); +AlCl
3 272, 369, 411; +AlCl
3/HCl 271, 304sh, 360, 408sh; +NaOAc 280, 325, 396; +NaOAc/H
3BO
3 266, 384. HR-MS (ESI) [M − H]
− calcd. for C
28H
29O
18: 653.1354, Found: 653.1344. LC-MS:
m/
z 655 [M + H]
+, 653 [M − H]
−,
m/
z 493 [M-162-H]
−,
m/
z 319 [M-42-132-162 + H]
+.
1H and
13C NMR, see
Table 1. Aglycone of
7 (gossypetin). HPLC (
tR): 4.91 min (solv. II). UV: λmax (nm) MeOH 261, 278, 304sh, 341, 381; +NaOMe decomp.; +AlCl
3 288, 329sh, 383, 470; +AlCl
3/HCl 272, 309sh, 370, 445; +NaOAc 275sh, 376; +NaOAc/H
3BO
3 305, 361.
3.4.8. Hibiscetin 3-O-glucoside-8-O-arabinoside (8)
TLC (Rf): 0.20 (BAW), 0.45 (BEW), 0.36 (15%HOAc); color UV (365 nm) dark purple, UV/NH
3 dark yellow. HPLC (
tR): 5.01 min (solv. I). UV: λmax (nm) MeOH 268, 369; + NaOMe decomp.; +AlCl
3 278, 443; +AlCl
3/HCl 277, 309, 368, 413; +NaOAc 279, 325, 420; +NaOAc/H
3BO
3 268, 393. HR-MS (ESI) [M − H]
− calcd. for C
26H
27O
18: 627.1197, Found 627.1178. LC-MS:
m/
z 629 [M + H]
+, 627 [M − H]
−,
m/
z 495 [M-132-H]
−,
m/
z 467 [M-162 + H]
+ and
m/
z 335 [M-162-132 + H]
+.
1H and
13C NMR, see
Table 1. Aglycone of
8 (hibiscetin). HPLC (
tR): 3.95 min (solv. II). UV: λmax (nm) MeOH 242sh, 299, 360; +NaOMe decomp.; +AlCl
3 259sh, 337, 406, 457sh; +AlCl
3/HCl 306, 373, 437sh; +NaOAc 308, 385; +NaOAc/H
3BO
3 308, 387.
3.4.9. Quercetin (9)
TLC (Rf): 0.76 (BAW), 0.76 (BEW), 0.01 (15%HOAc); color UV (365 nm) yellow, UV/NH3 yellow. HPLC (tR): 7.39 min (solv. II). UV: λmax (nm) MeOH 255, 273sh, 369; +NaOMe decomp.; +AlCl3 269, 449; +AlCl3/HCl 263, 296sh, 357, 425; +NaOAc 274, 327, 400; +NaOAc/H3BO3 258, 386. LC-MS: m/z 303 [M + H]+, 301 [M − H]−.
3.4.10. Quercetin 3-O-glucoside (Isoquercitrin, 10)
TLC (Rf): 0.67 (BAW), 0.73 (BEW), 0.23 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 yellow. HPLC (tR): 14.10 min (solv. I). UV: λmax (nm) MeOH 256, 266sh, 357; +NaOMe 275, 331, 409 (inc.); +AlCl3 275, 303sh, 434; +AlCl3/HCl 269, 297sh, 361, 400; +NaOAc 273, 325, 400; +NaOAc/H3BO3 261, 380. LC-MS: m/z 465 [M + H]+, 463 [M − H]−, m/z 303 [M-162 + H]+, 301 [M-162-H]−.
3.4.11. Quercetin 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside (11)
TLC (Rf): 0.51 (BAW), 0.68 (BEW), 0.84 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark yellow. HPLC (tR): 11.44 min (solv. I). UV: λmax (nm) MeOH 256, 265sh, 351; +NaOMe 273, 394 (inc.); +AlCl3 269, 404; +AlCl3/HCl 269, 301sh, 356, 397sh; +NaOAc 257, 264sh, 358, 407sh; +NaOAc/H3BO3 260, 368. LC-MS: m/z 725 [M − H]−, m/z 595 [M-132 + H]+, 593 [M-132-H]−, m/z 449 [M-132-146 + H]+, m/z 303 [M-132-146-146 + H]+. 1H NMR (800 MHz, DMSO-d6): δ 7.39 (1H, d, J = 2.4 Hz, H-2′), 7.30 (1H, dd, J = 2.4 and 8.4 Hz, H-6′), 6.89 (1H, d, J = 8.8 Hz, H-5′), 6.77 (1H, d, J = 1.6 Hz, H-8), 6.45 (1H, d, J = 2.4 Hz, H-6), 5.55 (1H, brs, 7-rhamnosyl H-1), 5.30 (1H, brs, 3-rhamnosyl H-1), 4.16 (1H, d, J = 8.0 Hz, xylosyl H-1), 4.07 (1H, brd, J = 3.2 Hz, 3-rhamnosyl H-2), 3.84 (1H, brs, 3-rhamnosyl H-3), 3.64 (1H, m, 7-rhamnosyl H-3), 3.61 (1H, m, 7-rhamnosyl H-5), 3.60 (1H, brd, J = 12.6 Hz, 7-rhamnosyl H-2), 3.43 (1H, m, xylosyl H-5b), 3.41 (1H, m, xylosyl H-4), 3.26 (1H, m, 3-rhamnosyl H-4), 3.19 (1H, m, 3-rhamnosyl H-5), 3.13 (1H, t, J = 9.6 Hz, 7-rhamnosyl H-4), 3.06 (1H, t, J = 8.8 Hz, xylosyl H-3), 2.95 (1H, t, J = 8.8 Hz, xylosyl H-2), 2.91 (1H, t, J = 11.2 Hz, xylosyl H-5a), 1.12 (3H, d, J = 6.2 Hz, 7-rhamnosyl CH3), 0.92 (3H, d, J = 6.2 Hz, 3-rhamnosyl CH3). 13C NMR (200 MHz, DMSO-d6): (quercetin) δ 157.6 (C-2), 134.5 (C-3), 178.0 (C-4), 160.9 (C-5), 99.4 (C-6), 161.7 (C-7), 94.5 (C-8), 156.0 (C-9), 105.6 (C-10), 120.1 (C-1′), 115.5 (C-2′), 145.4 (C-3′), 149.1 (C-4′), 115.6 (C-5′), 121.0 (C-6′); (3-O-rhamnose) δ 101.0 (C-1), 80.6 (C-2), 69.8 (C-3), 71.6 (C-4), 69.3 (C-5), 17.4 (C-6); (7-O-rhamnose) δ 98.4 (C-1), 70.1 (C-2), 70.3 (C-3), 71.7 (C-4), 70.2 (C-5), 17.9 (C-6); (2″-O-xylose) δ 106.5 (C-1), 73.8 (C-2), 76.2 (C-3), 70.3 (C-4), 65.7 (C-5).
3.4.12. Quercetin 3-O-rhamnoside-7-O-glucoside (12)
TLC (Rf): 0.31 (BAW), 0.52 (BEW), 0.64 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark yellow. HPLC (tR): 4.69 min (solv. I). UV: λmax (nm) MeOH 256, 265sh, 350; +NaOMe 272, 395 (inc.); +AlCl3 274, 435; +AlCl3/HCl 270, 297sh, 356, 397; +NaOAc 262, 395; +NaOAc/H3BO3 260, 369. LC-MS: m/z 611 [M + H]+, m/z 609 [M − H]−, m/z 465 [M-146 + H]+, m/z 303 [M-146-162 + H]+. 1H NMR (600 MHz, DMSO-d6): δ 7.31 (1H, d, J = 2.2 Hz, H-2′), 7.26 (1H, dd, J = 2.2 and 8.3 Hz, H-6′), 6.87 (1H, d, J = 8.3 Hz, H-5′), 6.73 (1H, d, J = 2.1 Hz, H-8), 6.44 (1H, d, J = 2.1 Hz, H-6), 5.27 (1H, d, J = 1.2 Hz, 3-rhamnosyl H-1), 5.06 (1H, d, J = 7.6 Hz, 7-glucosyl H-1), 3.97 (1H, dd, J = 1.5 and 3.1 Hz, 7-glucosyl H-4), 3.69 (1H, brd, J = 10.4 Hz, 7-glucosyl H-6b), 3.50 (1H, dd, J = 3.2 and 9.2 Hz, 3-rhamnosyl H-5), 3.44 (1H, m, 7-glucosyl H-6a), 3.43 (1H, m, 7-glucosyl H-5), 3.29 (1H, m, 7-glucosyl H-3), 3.25 (1H, m, 7-glucosyl H-2), 3.18 (1H, m, 3-rhamnosyl H-3), 3.17 (1H, m, 3-rhamnosyl H-2), 3.13 (1H, m, 3-rhamnosyl H-4), 0.81 (3H, d, J = 6.1 Hz, 3-rhamnosyl CH3). 13C NMR (150 MHz, DMSO-d6): (quercetin) δ 157.9 (C-2), 134.5 (C-3), 177.9 (C-4), 160.9 (C-5), 99.3 (C-6), 162.9 (C-7), 94.5 (C-8), 156.1 (C-9), 105.7 (C-10), 120.5 (C-1′), 115.8 (C-2′), 145.3 (C-3′), 148.7 (C-4′), 115.5 (C-5′), 121.2 (C-6′); (3-O-rhamnose) δ 101.8 (C-1), 70.0 (C-2), 70.6 (C-3), 71.2 (C-4), 70.4 (C-5), 17.5 (C-6); (7-O-glucose) δ 99.9 (C-1), 73.1 (C-2), 76.4 (C-3), 69.6 (C-4), 77.2 (C-5), 60.6 (C-6).
3.4.13. Kaempferol (13)
TLC (Rf): 0.95 (BAW), 0.95 (BEW), 0.01 (15%HOAc); color UV (365 nm) yellow, UV/NH3 yellow. HPLC (tR): 11.47 min (solv. II). UV: λmax (nm) MeOH 268, 367; +NaOMe decomp.; +AlCl3 270, 304sh, 350, 427; +AlCl3/HCl 269, 301sh, 349, 427; +NaOAc 276, 313sh, 396; +NaOAc/H3BO3 268, 370. LC-MS: m/z 287 [M + H]+, 285 [M − H]−.
3.4.14. Kaempferol 3-O-glucoside (Astragalin, 14)
TLC (Rf): 0.80 (BAW), 0.85 (BEW), 0.36 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark greenish yellow. HPLC (tR): 20.61 min (solv. I). UV: λmax (nm) MeOH 265, 349; +NaOMe 275, 323, 401 (inc.); +AlCl3 268, 303, 350, 402sh; +AlCl3/HCl 269, 300, 348, 396sh; +NaOAc 272, 304, 373; +NaOAc/H3BO3 264, 359. LC-MS: m/z 449 [M + H]+, 447 [M − H]−, m/z 287 [M-162 + H]+.
3.4.15. Kaempferol 7-O-rhamnoside (15)
TLC (Rf): 0.85 (BAW), 0.93 (BEW), 0.08 (15%HOAc); color UV (365 nm) yellow, UV/NH3 yellow. HPLC (tR): 23.86 min (solv. I). UV: λmax (nm) MeOH 269, 370; +NaOMe decomp.; +AlCl3 271, 303sh, 361, 422; +AlCl3/HCl 270, 301sh, 355, 423; +NaOAc 279, 316, 394; +NaOAc/H3BO3 270, 375. LC-MS: m/z 433 [M + H]+, 431 [M − H]−, m/z 287 [M-146 + H]+.
3.4.16. Kaempferol 3,7-di-O-rhamnoside (16)
TLC (Rf): 0.79 (BAW), 0.81 (BEW), 0.71 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark yellow. HPLC (tR): 18.06 min (solv. I). UV: λmax (nm) MeOH 266, 342; +NaOMe 277, 383 (inc.); +AlCl3 276, 301, 347, 401; +AlCl3/HCl 276, 299, 342, 399; +NaOAc 269, 391; +NaOAc/H3BO3 268, 347. LC-MS: m/z 579 [M + H]+, 577 [M − H]−, m/z 433 [M-146 + H]+, 431 [M-146-H]−, m/z 287 [M-146-146 + H]+, 285 [M-146-146-H]−.
3.4.17. Kaempferol 3-O-glucoside-7-O-rhamnoside (17)
TLC (Rf): 0.52 (BAW), 0.70 (BEW), 0.64 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark greenish yellow. HPLC (tR): 10.67 min (solv. I). UV: λmax (nm) MeOH 265, 350; +NaOMe 275, 390 (inc.); +AlCl3 275, 299sh, 351, 398; +AlCl3/HCl 275, 298sh, 348, 395; +NaOAc 268, 398; +NaOAc/H3BO3 266, 356. LC-MS: m/z 595 [M + H]+, 593 [M − H]−, m/z 447 [M-146-H]−, 433 [M-162 + H]+, m/z 287 [M-162-146 + H]+.
3.4.18. Kaempferol 3-O-glucosyl-(1→2)-rhamnoside-7-O-rhamnoside (18)
TLC (Rf): 0.57 (BAW), 0.64 (BEW), 0.85 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark yellow. HPLC (tR): 13.61 min (solv. I). UV: λmax (nm) MeOH 265, 339; +NaOMe 273, 379 (inc.); +AlCl3 267, 299, 345, 400sh; +AlCl3/HCl 268, 297sh, 340, 397sh; +NaOAc 265, 350; +NaOAc/H3BO3 266, 352. LC-MS: m/z 739 [M − H]−, m/z 593 [M-146-H]−, m/z 433 [M-146-162 + H]+, m/z 287 [M-146-146-162 + H]+. 1H NMR (600 MHz, DMSO-d6): δ 7.80 (2H, d, J = 8.8 Hz, H-2′,6′), 6.92 (2H, d, J = 8.8 Hz, H-3′,5′), 6.78 (1H, d, J = 2.0 Hz, H-8), 6.44 (1H, d, J = 2.1 Hz, H-6), 5.55 (1H, brs, 3-rhamnosyl H-1), 5.54 (1H, brs, 7-rhamnosyl H-1), 4.23 (1H, d, J = 7.9 Hz, 2″-glucosyl H-1), 4.08 (1H, brd, J = 2.3 Hz, 2″-glucosyl H-4), 3.83 (1H, brs, 3-rhamnosyl H-3), 3.62 (1H, dd, J = 3.3 and 9.3 Hz, 3-rhamnosyl H-4), 3.54 (1H, brd, J = 8.5 Hz, 7-rhamnosyl H-3), 3.51 (1H, m, 2″-glucosyl H-6b), 3.43 (1H, m, 7-rhamnosyl H-5), 3.41 (1H, m, 2″-glucosyl H-6a), 3.40 (1H, m, 3-rhamnosyl H-2), 3.30 (1H, m, 3-rhamnosyl H-5), 3.29 (1H, m, 7-rhamnosyl H-4), 3.16 (1H, m, 2″-glucosyl H-3), 3.12 (1H, m, 2″-glucosyl H-5), 2.99 (1H, m, 7-rhamnosyl H-2), 2.98 (1H, m, 2″-glucosyl H-2), 1.11 (3H, d, J = 6.2 Hz, 7-rhamnosyl CH3), 0.87 (3H, d, J = 6.2 Hz, 3-rhamnosyl CH3). 13C NMR (150 MHz, DMSO-d6): (kaempferol) δ 157.6 (C-2), 134.8 (C-3), 177.9 (C-4), 161.0 (C-5), 99.4 (C-6), 161.7 (C-7), 94.6 (C-8), 156.1 (C-9), 105.8 (C-10), 120.1 (C-1′), 130.7 (C-2′), 115.5 (C-3′), 160.4 (C-4′), 115.5 (C-5′), 130.7 (C-6′); (3-O-rhamnose) δ 101.0 (C-1), 81.2 (C-2), 70.1 (C-3), 71.6 (C-4), 69.3 (C-5), 17.9 (C-6); (7-O-rhamnose) δ 98.5 (C-1), 70.2 (C-2), 70.4 (C-3), 71.7 (C-4), 70.3 (C-5), 17.4 (C-6); (2″-O-glucose) δ 106.1 (C-1), 73.9 (C-2), 76.3 (C-3), 69.8 (C-4), 76.6 (C-5), 60.5 (C-6).
3.4.19. Kaempferol 3-O-xylosyl-(1→2)-rhamnoside (19)
TLC (Rf): 0.85 (BAW), 0.91 (BEW), 0.56 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark greenish yellow. HPLC (tR): 14.78 min (solv. I). UV: λmax (nm) MeOH 265, 337; +NaOMe 273, 322, 387 (inc.); +AlCl3 274, 303, 348, 390; +AlCl3/HCl 275, 300, 340, 392; +NaOAc 274, 321, 381; +NaOAc/H3BO3 266, 344. LC-MS: m/z 563 [M − H]−, m/z 433 [M-132 + H]+, m/z 287 [M-132-146 + H]+. 1H NMR (800 MHz, DMSO-d6): δ 7.76 (2H, d, J = 8.8 Hz, H-2′,6′), 6.92 (2H, d, J = 8.8 Hz, H-3′,5′), 6.34 (1H, brs, H-8), 6.15 (1H, brs, H-6), 5.38 (1H, brs, rhamnosyl H-1), 4.18 (1H, d, J = 7.2 Hz, xylosyl H-1), 4.01 (1H, brs, rhamnosyl H-2), 3.53 (1H, brd, J = 5.6 Hz, rhamnosyl H-3), 3.51 (1H, brd, J = 4.8 Hz, xylosyl H-5b), 3.42 (1H, dd, J = 4.8 and 8.8 Hz, xylosyl H-4), 3.20 (1H, m, rhamnosyl H-5), 3.11 (1H, t, J = 8.8 Hz, rhamnosyl H-4), 3.07 (1H, t, J = 8.8 Hz, xylosyl H-3), 2.96 (1H, t, J = 8.8 Hz, xylosyl H-2), 2.93 (1H, t, J = 11.2 Hz, xylosyl H-5a), 0.87 (3H, d, J = 6.4Hz, rhamnosyl CH3). 13C NMR (200 MHz, DMSO-d6): (kaempferol) δ 156.7 (C-2), 134.1 (C-3), 177.5 (C-4), 161.2 (C-5), 99.1 (C-6), 156.6 (C-7), 93.9 (C-8), 152.2 (C-9), 103.2 (C-10), 120.3 (C-1′), 130.4 (C-2′), 115.4 (C-3′), 161.1 (C-4′), 115.4 (C-5′), 130.4 (C-6′); (3-O-rhamnose) δ 100.7 (C-1), 80.5 (C-2), 70.3 (C-3), 71.6 (C-4), 69.3 (C-5), 17.4 (C-6); (2″-O-xylose) δ 106.4 (C-1), 73.7 (C-2), 76.2 (C-3), 70.3 (C-4), 65.8 (C-5).
3.4.20. Kaempferol 3-O-xylosyl-(1→2)-rhamnoside-7-O-rhamnoside (20)
TLC (Rf): 0.44 (BAW), 0.67 (BEW), 0.85 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 dark yellow. HPLC (tR): 13.86 min (solv. I). UV: λmax (nm) MeOH 265, 340; +NaOMe 273, 378 (inc.); +AlCl3 275, 300, 344, 397; +AlCl3/HCl 275, 297sh, 340, 395; +NaOAc 266, 385; +NaOAc/H3BO3 265, 344. LC-MS: m/z 711 [M + H]+, 709 [M − H]−, m/z 579 [M-132 + H]+, 577 [M-132-H]−, m/z 433 [M-132-146 + H]+, m/z 287 [M-132-146-146 + H]+. 1H NMR (800 MHz, DMSO-d6): δ 7.81 (2H, d, J = 8.8 Hz, H-2′,6′), 6.94 (2H, d, J = 8.8 Hz, H-3′,5′), 6.79 (1H, d, J = 2.4 Hz, H-8), 6.46 (1H, d, J = 1.6 Hz, H-6), 5.55 (1H, brs, 7-rhamnosyl H-1), 5.38 (1H, brs, 3-rhamnosyl H-1), 4.18 (1H, d, J = 8.0 Hz, xylosyl H-1), 4.03 (1H, dd, J = 1.6 and 3.6 Hz, 3-rhamnosyl H-2), 3.81 (1H, brs, 3-rhamnosyl H-3), 3.63 (1H, m, 7-rhamnosyl H-3), 3.62 (1H, m, 7-rhamnosyl H-5), 3.56 (1H, m, 7-rhamnosyl H-2), 3.51 (1H, dd, J = 5.6 and 11.2 Hz, xylosyl H-5b), 3.43 (1H, dd, J = 5.6 and 9.2 Hz, xylosyl H-4), 3.30 (1H, t, J = 9.6 Hz, 3-rhamnosyl H-4), 3.22 (1H, m, 3-rhamnosyl H-5), 3.12 (1H, t, J = 8.8 Hz, 7-rhamnosyl H-4), 3.07 (1H, t, J = 9.6 Hz, xylosyl H-3), 2.96 (1H, t, J = 8.8 Hz, xylosyl H-2), 2.93 (1H, t, J = 11.2 Hz, xylosyl H-5a), 1.12 (3H, d, J = 6.4 Hz, 7-rhamnosyl CH3), 0.89 (3H, d, J = 6.4 Hz, 3-rhamnosyl CH3). 13C NMR (200 MHz, DMSO-d6): (kaempferol) δ 157.6 (C-2), 134.6 (C-3), 178.0 (C-4), 161.0 (C-5), 99.5 (C-6), 161.7 (C-7), 94.7 (C-8), 156.1 (C-9), 105.7 (C-10), 120.1 (C-1′), 130.6 (C-2′), 115.5 (C-3′), 160.4 (C-4′), 115.5 (C-5′), 130.6 (C-6′); (3-O-rhamnose) δ 100.9 (C-1), 80.5 (C-2), 69.8 (C-3), 71.6 (C-4), 69.3 (C-5), 17.4 (C-6); (7-O-rhamnose) δ 98.4 (C-1), 70.1 (C-2), 70.4 (C-3), 71.7 (C-4), 70.2 (C-5), 17.9 (C-6); (2″-O-xylose) δ 106.4 (C-1), 73.7 (C-2), 76.3 (C-3), 70.3 (C-4), 65.8 (C-5).
3.4.21. Myricetin 3-O-glucoside (21)
TLC (Rf): 0.40 (BAW), 0.61 (BEW), 0.18 (15%HOAc); color UV (365 nm) dark purple, UV/NH3 yellow. HPLC (tR): 8.77 min (solv. I). UV: λmax (nm) MeOH 257, 264sh, 360; +NaOMe decomp.; +AlCl3 272, 431; +AlCl3/HCl 283, 309, 365sh, 402; +NaOAc 272, 325, 406; +NaOAc/H3BO3 260, 300, 382. LC-MS: m/z 481 [M + H]+, 479 [M − H]−, m/z 319 [M-162 + H]+.
3.4.22. Cyanidin 3-O-glucoside (Chrysanthemin, 22)
HPLC (tR): 4.19 min (solv. I). UV: λmax (nm) 0.01%HCl-MeOH 277, 332, 528; +AlCl3 275, 537; E440/Emax = 26.5%. LC-MS: m/z 449 [M]+, m/z 287 [M-162]+.