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Solvents were removed in a vacuum to obtain a crude extract
(1050 g), which was suspended in water and partitioned with
EtOAc and n-BuOH to yield an EtOAc part (185 g), an n-BuOH
part (390 g) and an aqueous extract (382 g). The n-BuOH part
(350 g) was subjected to silica gel column chromatography
using gradient CH2Cl2–CH3OH–H2O (30 : 1 : 1–1 : 1 : 1) to
obtain 12 fractions based on TLC analysis. Fraction 6 (8 g) was
submitted to silica gel column chromatography and eluted with
a CH2Cl2–CH3OH system (15 : 1–1 : 1) to yield six sub-fractions.
Subfraction 6-5 (2.8 g) was further separated by Sephadex LH-20
column chromatography (CH2Cl2–CH3OH, 1 : 1), and the
resultant fractions were further puried by RP-18 HPLC
(CH3OH–H2O, 78 : 22) to obtain compounds 2 (11 mg) and 5
(13 mg). Fraction 8 (4.2 g) was separated by ash ODS column
chromatography (CH3OH : H2O, 30 : 70–80 : 20) to yield six
subfractions. Sub-fraction 8-4 (600 mg) was further puried by
RP-18 HPLC (CH3OH : H2O, 70 : 30) to yield compounds 3
(13 mg), 4 (9 mg), 6 (12 mg), and 7 (11 mg). Compound 1 (8 mg)
was also obtained from subfraction 8-6 (800 mg) by RP-18 HPLC
(CH3OH : H2O, 65 : 35). Fraction 11 (75.9 g) was subjected to
ash ODS column chromatography (CH3OH : H2O, 10 : 90–
100 : 0) to afford nine subfractions, and subfraction 11-5
(150 mg) was puried by RP-18 HPLC with CH3OH–H2O
Compound 4. 16-O-Acetyl-aesculioside G12; white amor-
phous powder (methyl alcohol); [a]2D0 ¼ +23.0 (c 0.07, CH3OH);
IR (KBr) nmax 1028, 1720, 1743, 2928, 2953, 3420 cmꢀ1; 1H-NMR
(C5D5N, 600 MHz) and 13C-NMR (C5D5N, 150 MHz) are shown in
Table 2; HR-ESI-MS (positive-ion model) m/z 1203.5925 [M +
Na]+ (calcd for C60H92O23Na, 1203.5927).
Compound
5.
3-O-[a-L-Arabinofuranosyl(1/3)]-b-D-gal-
actopyranosyl(1/2)-b-D-60-methyl-glucuronic acid-21-O-(30000,40000
-
O-diangeloyl)-b-D-fucopyranosyl-28-O-acetyl-3b,16a,21b,22a,28-
pentahydroxy-olean-12-ene; white amorphous powder (methyl
alcohol); [a]2D0 ¼ +23.1 (c 0.09, CH3OH); IR (KBr) nmax 1041,
1071, 1630, 1723, 2927, 3427 cmꢀ1; 1H-NMR (C5D5N, 600 MHz)
and 13C-NMR (C5D5N, 150 MHz) are shown in Table 1; HR-ESI-
MS (positive-ion model) m/z 1349.6426 [M + Na]+ (calcd for
C
66H102O27Na, 1349.6506).
Compound 6. 60-Methylester-O-xanifolia-Y2; white amor-
phous powder (methyl alcohol); [a]2D0 ¼ +24.5 (c 0.05, CH3OH);
IR (KBr) nmax 1039, 1075, 1720, 2922, 3400 cmꢀ1 1H-NMR
;
(C5D5N, 600 MHz) and 13C-NMR (C5D5N, 150 MHz) are shown
in Table 1; HR-ESI-MS (positive-ion model) m/z 1193.5719 [M +
Na]+ (calcd for C58H90O24Na, 1193.5720).
Compound 7. 60-Methylester-O-xanifolia-Y8; white amor-
phous powder (methyl alcohol); [a]2D0 ¼ +71.7 (c 0.06, CH3OH);
IR (KBr) nmax 1027, 1082, 1720, 2921, 3400 cmꢀ1 1H-NMR
;
(75 : 25) solvent system to yield compound
9 (10 mg).
(C5D5N, 300 MHz) and 13C-NMR (C5D5N, 150 MHz) are shown
in Table 1; HR-ESI-MS (positive-ion model) m/z 1177.5759 [M +
Na]+ (calcd for C58H90O23Na, 1177.5771).
Compound 8 (3 g) was recrystallized from subfraction 11-7
(10 g) in methanol–dichloromethane system. Fraction 12 (7.9 g)
was subjected to silica gel column chromatography and eluted
with a CH2Cl2–CH3OH system (100 : 5–100 : 70) to obtain eight
subfractions. Subfraction 12-2 (190 mg) was puried with using
Sephadex LH-20 column chromatography (CH3OH : H2O,
65 : 35), and the resultant fractions were further puried using
RP-18 HPLC with a CH3OH : H2O : CF3COOH (70 : 30 : 0.03)
solvent system to obtain compound 10 (13 mg).
Compound 8. Xanifolia Y: white acicular crystal (methyl
alcohol), 1H-NMR (C5D5N, 400 MHz) dH 0.85 (3H, s, H-25), 1.02
(3H, s, H-26), 1.13 (3H, s, H-29), 1.17 (3H, s, H-24), 1.30 (3H, s, H-
23), 1.35 (3H, s, H-30), 1.76 (3H, s, 22-Ang-H-5), 1.88 (3H, s, H-
27), 1.98 (3H, d, J ¼ 7.1 Hz, 22-Ang-H-4), 2.04 (3H, s, 21-Ang-
H-5), 2.11 (3H, d, J ¼ 7.2 Hz, 21-Ang-H-4), 3.27 (1H, dd, J ¼
11.6, 4.1 Hz, H-3), 3.52 (1H, d, J ¼ 10.6 Hz, H-28a), 3.78 (1H, d,
J ¼ 10.6 Hz, H-28b), 4.94 (1H, d, J ¼ 7.6 Hz, H-10), 5.36 (1H, d, J ¼
7.7 Hz, H-100), 5.62 (1H, brs, H-12), 5.80 (1H, d, J ¼ 7.1 Hz, 22-
Ang-H-3), 5.99 (1H, d, J ¼ 7.2 Hz, 21-Ang-H-3), 6.07 (1H, d, J ¼
2.0 Hz, H-1000), 6.36 (1H, d, J ¼ 10.2 Hz, H-22), 6.74 (1H, d, J ¼
10.2 Hz, H-21); 13C-NMR (C5D5N, 100 MHz) dC 39.6 (C-1), 26.6
(C-2), 89.9 (C-3), 38.9 (C-4), 55.5 (C-5), 18.8 (C-6), 36.6 (C-7), 40.1
(C-8), 47.1 (C-9), 36.9 (C-10), 23.9 (C-11), 125.4 (C-12), 143.6 (C-
13), 48.3 (C-14), 67.5 (C-15), 73.4 (C-16), 47.6 (C-17), 41.4 (C-
18), 46.8 (C-19), 36.3 (C-20), 78.8 (C-21), 73.6 (C-22), 27.8
(C-23), 16.7 (C-24), 15.9 (C-25), 17.5 (C-26), 21.2 (C-27), 63.0 (C-
28), 29.4 (C-29), 20.2 (C-30), 105.1 (C-10), 78.6 (C-20), 86.2 (C-
30), 71.7 (C-40), 77.2 (C-50), 172.1 (C-60), 104.7 (C-100), 73.3 (C-200),
75.0 (C-300), 69.7 (C-400), 76.6 (C-500), 61.8 (C-600), 111.1 (C-1000), 83.5
(C-2000), 77.6 (C-3000), 85.4 (C-4000), 62.3 (C-5000), 167.7 (C-21-Ang-1),
128.8 (C-21-Ang-2), 137.3 (C-21-Ang-3), 15.6 (C-21-Ang-4), 20.6
(C-21-Ang-5), 168.1 (C-22-Ang-1), 129.0 (C-22-Ang-2), 136.5 (C-
22-Ang-3), 15.7 (C-22-Ang-4), 20.9 (C-22-Ang-5).
Compound 1. 3-O-(30-O-Angeloyl)-b-D-glucopyranosyl-28-O-
[a-L-rhamnopyranosyl(1/2)]-b-D-glucopyranosyl-3b,21b,22a,28-
tetrahydroxy-olean-12-ene; white amorphous powder (CH3OH);
[a]2D0 ¼ +137.8 (c 0.04, CH3OH); IR (KBr) nmax 1048, 1719, 2923,
1
3423 cmꢀ1; H NMR (C5D5N, 600 MHz) and 13C NMR (C5D5N,
150 MHz) spectral data were given in Table 1; HR-ESI-MS
(positive-ion model) m/z 1049.5656 [M + Na]+ (calcd for
C53H86O19Na, 1049.5661).
Compound 2. 3-O-[a-L-Arabinofuranosyl(1/3)]-b-D-gal-
actopyranosyl(1/2)-b-D-60-n-butyl-glucuronic acid-21-O-epoxy-
angeloyl-22-O-angeloyl-3b,16a,21b,22a,28-pentahydroxy-olean-
12-ene; white amorphous powder (CH3OH); [a]2D0 ¼ +30.9
(c 0.07, CH3OH); IR (KBr) nmax 1082, 1738, 2929, 2959,
1
3422 cmꢀ1; H-NMR (C5D5N, 300 MHz) and 13C-NMR (C5D5N,
150 MHz) are shown in Table 2; HR-ESI-MS (positive-ion
model) m/z 1219.6238 [M + Na]+ (calcd for C61H96O23Na,
1219.6240).
Compound 3. 60-Methylester-O-xanifolia-Y5, white amor-
phous powder (methyl alcohol); [a]2D0 ¼ +56.6 (c 0.09, CH3OH);
Compound 9. Xanifolia ACH-Y: white amorphous powder
(methyl alcohol), 1H-NMR (C5D5N, 600 MHz); 13C-NMR (C5D5N,
100 MHz) dH 1.00 (3H, s, H-25), 1.08 (3H, s, H-24), 1.12 (3H, s,
H-29), 1.13 (3H, s, H-26), 1.26 (3H, s, H-23), 1.37 (3H, s, H-30),
1.76 (3H, s, 22-Ang-H-5), 1.88 (3H, s, H-27), 1.97 (3H, d, J ¼
IR (KBr) nmax 1077, 1680, 2927, 3426 cmꢀ1 1H-NMR (C5D5N,
;
600 MHz) and 13C-NMR (C5D5N, 150 MHz) spectral data were
shown in Table 2; HR-ESI-MS (positive-ion model) m/z
1045.5294 [M + Na]+ (calcd for C53H82O19Na, 1045.5348).
27444 | RSC Adv., 2016, 6, 27434–27446
This journal is © The Royal Society of Chemistry 2016