858
Vol. 57, No. 8
given in Table 1. 13C-NMR (125 MHz, CD3OD) dC: given in Table 1. Posi-
tive-ion FAB-MS m/z: 447 (MꢁNa)ꢁ.
(6.40 g), Fr. 6 (11.00 g), Fr. 7 (5.40 g), Fr. 8 (4.00 g), Fr. 9 (7.10 g), Fr. 10
(5.80 g), Fr. 11 (6.10 g), and Fr. 12 (17.10 g)] as reported previously.17,18)
Fraction 4 (1.80 g) was subjected to reversed-phase silica gel CC
[70 g, MeOH–H2O (15 : 85→80 : 20, v/v)→MeOH] and HPLC [MeOH–
H2O (20 : 80—30 : 70, v/v)] to give everlastoside I (4, 22.0 mg, 0.0055%) to-
gether with everlastoside A (2.3 mg, 0.0005%) and 7-hydroxy-5-methoxy-
Everlastoside G (2): A white powder, [a]D24 ꢀ30.3° (cꢂ0.61, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C19H32O13Na (MꢁNa)ꢁ
491.1741; Found 491.1745. UV [lmax (log e), MeOH]: 217 (4.82) nm. IR
(KBr, cmꢀ1): 3568, 1718, 1686, 1508, 1458, 1066. 1H-NMR (500 MHz,
CD3OD) d: given in Table 1. 13C-NMR (125 MHz, CD3OD) dC: given in
Table 1. Positive-ion FAB-MS m/z: 491 (MꢁNa)ꢁ.
phthalide 7-O-b-D-glucopyranoside (476.8 mg, 0.12%).17,18) Fraction
5
(4.00 g) was purified by reversed-phase silica gel CC [300 g, MeOH–H2O
(15 : 85→80 : 20, v/v)→MeOH] and HPLC [MeOH–H2O (15 : 85—45 : 55,
v/v)] to furnish 7-O-(b-D-glucopyranosyloxy)-5-hydroxy-1(3H)-isobenzofu-
ranone (9, 93.0 mg, 0.023%) and everlastoside I (4, 10.0 mg, 0.0025%) to-
gether with (2S)-helichrysin (320.0 mg, 0.069%), (2R)-helichrysin (18.0 mg,
0.0045%), naringenin 7-O-b-D-glucopyranoside (21.5 mg, 0.0053%), api-
genin 7-O-b-D-glucopyranoside (10.0 mg, 0.0025%), luteolin 7-O-b-D-glu-
copyranoside (10.0 mg, 0.0025%), luteolin 3ꢄ-O-b-D-glucopyranoside
(5.3 mg, 0.0013%), kaempferol 3-O-b-D-glucopyranoside (2.30 g, 0.58%),
tortoside B (10.0 mg, 0.0025%), 7-hydroxy-5-methoxyphthalide 7-O-b-D-
glucopyranoside (150.0 mg, 0.037%), scopolin (68.0 mg, 0.017%), undulato-
side A (11.0 mg, 0.0017%), 4-(3ꢄ-glucopyranosyloxy-4ꢄ-hydroxyphenyl)-3-
buten-2-one (7a, 11.3 mg, 0.0023%), syringin (15.0 mg, 0.0037%), dihy-
drosyringin (7.0 mg, 0.0013%), and eugenyl b-D-glucopyranoside (11.8 mg,
0.0029%).17) Fraction 7 (5.40 g) was subjected by reversed-phase silica gel
CC [300 g, MeOH–H2O (15 : 85→70 : 30, v/v)→MeOH] and HPLC
[MeOH–H2O (10 : 90—40 : 60, v/v)] to furnish 9 (65.3 mg, 0.016%) and
Everlastoside H (3): A white powder, [a]D28 ꢀ35.2° (cꢂ0.67, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C21H28O14Na (MꢁNa)ꢁ
527.1376; Found 527.1371. UV [lmax (log e), MeOH]: 216 (4.50), 257
1
(4.20) nm. IR (KBr, cmꢀ1): 3568, 1719, 1612, 1508, 1458, 1343, 1066. H-
NMR (500 MHz, CD3OD) d: given in Table 2. 13C-NMR (125 MHz,
CD3OD) dC: given in Table 2. Positive-ion FAB-MS m/z: 527 (MꢁNa)ꢁ.
Everlastoside I (4): A white powder, [a]D23 ꢀ32.0° (cꢂ0.67, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C16H25O7 (MꢁH)ꢁ
329.1600; Found 329.1596. IR (KBr, cmꢀ1): 3568, 1509, 1458, 1375, 1071.
1H-NMR (500 MHz, CD3OD) d: given in Table 3. 13C-NMR (125 MHz,
CD3OD) dC: given in Table 3. Positive-ion FAB-MS m/z: 329 (MꢁH)ꢁ, 351
(MꢁNa)ꢁ.
Everlastoside J (5): A white powder, [a]D26 ꢀ89.3° (cꢂ0.30, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C21H32O11Na (MꢁNa)ꢁ
483.1842; Found 483.1846. IR (KBr, cmꢀ1): 3568, 1508, 1458, 1341, 1071.
1H-NMR (500 MHz, CD3OD) d: given in Table 3. 13C-NMR (125 MHz,
CD3OD) dC: given in Table 3. Positive-ion FAB-MS m/z: 461 (MꢁH)ꢁ, 483
(MꢁNa)ꢁ.
everlastoside
J (5, 5.0 mg, 0.0012%) together with everlastosides B
(12.9 mg, 0.0032%), C (16.9 mg, 0.0042%), D (6.2 mg, 0.0015%), and E
(23.0 mg, 0.0060%), (2S)-helichrysin (223.0 mg, 0.055%), (2R)-helichrysin
(17.0 mg, 0.0042%), chalconaringenin 2ꢄ-O-b-D-glucopyranoside (305.5 mg,
0.076%), quercetin 3-O-b-D-glucopyranoside (40.0 mg, 0.010%), (7R,8S)-
dihydrodehydrodiconiferyl alcohol 4-O-b-D-glucopyranoside (10.0 mg,
0.0025%), oricinol b-D-glucopyranoside (12.2 mg, 0.0035%), phenethyl al-
cohol b-D-xylopyranosyl-(1→6)-b-D-glucopyranoside (6.2 mg, 0.0015%),
icariside D1 (90.0 mg, 0.017%), and adenosine (22.0 mg, 0.0055%).17,18)
Fraction 8 (4.00 g) was subjected to reversed-phase silica gel CC [300 g,
MeOH–H2O (15 : 85→60 : 40, v/v)→MeOH] and HPLC [MeOH–H2O
(18 : 82—40 : 60, v/v) or CH3CN–H2O (8 : 92—11 : 89, v/v)] to give everlas-
toside G (2, 16.4 mg, 0.0041%) and licoagroside B (10, 6.0 mg, 0.0015%)
together with (2S)-helichrysin (20.0 mg, 0.0050%), (2R)-helichrysin
Everlastoside K (6): A white powder, [a]D25 ꢀ34.2° (cꢂ1.04, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C22H34O12Na (MꢁNa)ꢁ
513.1948; Found 513.1937. IR (KBr, cmꢀ1): 3568, 1508, 1458, 1071. 1H-
NMR (500 MHz, CD3OD) d: given in Table 3. 13C-NMR (125 MHz,
CD3OD) dC: given in Table 3. Positive-ion FAB-MS m/z: 491 (MꢁH)ꢁ, 513
(MꢁNa)ꢁ.
Everlastoside L (7): A white powder, [a]D22 ꢁ34.5° (cꢂ0.25, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C19H32O13Na (MꢁNa)ꢁ
525.1373; Found 525.1367. UV [lmax (log e), MeOH]: 329 (4.07) nm. IR
(KBr, cmꢀ1): 3433, 1716, 1686, 1655, 1541, 1509, 1458, 1073. 1H-NMR
(500 MHz, CD3OD) d: given in Table 4. 13C-NMR (125 MHz, CD3OD) dC:
given in Table 4. Positive-ion FAB-MS m/z: 525 (MꢁNa)ꢁ.
Everlastoside M (8): A white powder, [a]D26 ꢀ80.8° (cꢂ0.50, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C22H26O12Na (MꢁNa)ꢁ
505.1322; Found 505.1327. UV [lmax (log e), MeOH]: 258 (3.87) nm. IR
(KBr, cmꢀ1): 3415, 1718, 1698, 1634, 1607, 1516, 1499, 1254, 1169, 1080.
1H-NMR (500 MHz, CD3OD) d: given in Table 4. 13C-NMR (125 MHz,
CD3OD) dC: given in Table 4. Positive-ion FAB-MS m/z: 505 (MꢁNa)ꢁ.
Alkaline and Acid Hydrolysis of Everlastosides F (1), G (2) A solu-
tion of 1 or 2 (each 6.0 mg) in 10% aqueous KOH–50% aqueous 1,4-dioxane
(1 : 1, v/v, 1.0 ml) was stirred at 37 °C for 3 h. The reaction mixture was neu-
tralized with Dowex HCR W2 (Hꢁ form) and the resin was removed by fil-
tration. Evaporation of the solvent from the filtrate under reduced pressure
yielded a residue. A part of residue was dissolved in (CH2)2Cl2 (2.0 ml) and
the solution was treated with p-nitrobenzyl-N-Nꢄ-diisopyopylisourea
(10 mg), then the whole was stirred at 80 °C for 1 h. The reaction mixture
was subjected to HPLC analysis [column: YMC-Pack ODS-A, 250ꢆ4.6 mm
i.d.; mobile phase: MeOH–H2O (70 : 30, v/v); detection: UV (254 nm); flow
rate: 0.9 ml/min] to identify the p-nitrobenzyl ester of angelic acid (tR
16.0 min), respectively. The rest of residue in 1 M HCl (1.0 ml) was heated at
80 °C for 1 h. After cooling, the reaction mixture was neutralized with Am-
berlite IRA-400 (OHꢀ form) and then the resin was removed by filtration.
Removal of the solvent from the filtrate under reduced pressure, the residue
was separated by Sep-Pak C18 cartridge column (H2O→MeOH). The H2O-
eluted fraction was subjected to HPLC analysis under the following condi-
tions: HPLC column, Kaseisorb LC NH2-60-5, 250ꢆ4.6 mm i.d. (Tokyo
Kasei Co., Ltd., Tokyo, Japan); detection, optical rotation [Shodex OR-2
(Showa Denko Co., Ltd., Tokyo, Japan); mobile phase, CH3CN-H2O
(85 : 15, v/v); flow rate 0.8 ml/min]. Identification of D-apiose18,22,23) (i, from
2) and D-glucose (ii, from 1 and 2) present in the aqueous layer was carried
out by comparison of their retention times and optical rotations with those of
authentic samples. tR: (i) 6.6 min (positive optical rotation) and (ii) 13.9 min
(positive optical rotation), respectively.
(30.0 mg, 0.0075%), helicioside
A (6.0 mg, 0.0015%), (2R,3R)-dihy-
drokaempferol 7-O-b-D-glucopyranoside (40.0 mg, 0.010%), chalconarin-
genin 2ꢄ-O-b-D-glucopyranoside (25.0 mg, 0.0057%), quercetin 3-O-b-D-
glucopyranoside (70.0 mg, 0.018%), benzyl alcohol b-D-xylopyranosyl-
(1→6)-b-D-glucopyranoside (5.0 mg, 0.0012%), icariside F2 (19.0 mg,
0.0048%), and phenethyl alcohol b-D-xylopyranosyl-(1→6)-b-D-glucopyra-
noside (7.2 mg, 0.0018%).17) Fraction 9 (7.10 g) was subjected by reversed-
phase silica gel CC [300 g, MeOH–H2O (15 : 85→60 : 40, v/v)→MeOH] and
HPLC [MeOH–H2O (9 : 92—35 : 65, v/v)] to furnish everlastosides F (1,
30.0 mg, 0.0075%), H (3, 10.0 mg, 0.0025%), and K (6, 16.0 mg, 0.0040%)
together with scutellarein 7-O-b-D-glucopyranoside (7.0 mg, 0.0017%),
3ꢄ-methylchrysoeriol 7-O-b-D-glucopyranoside (13.0 mg, 0.0033%),
kaempferol 3-O-b-D-glucopyranoside (13.0 mg, 0.0032%), kaempferol 3-
O-b-D-glucopyranosyl-(1→3)-b-D-glucopyranoside (17.0 mg, 0.0040%),
quercetin 3-O-b-D-glucopyranoside (13.3 mg, 0.0033%), aureusidin 6-O-b-
D-glucopyranoside (10.0 mg, 0.0025%), and benzoyl b-D-glucopyranosyl-
(1→6)-b-D-glucopyranoside (36.0 mg, 0.0090%).17) Fraction 10 (5.10 g)
was subjected to reversed-phase silica gel CC [200 g, MeOH–H2O
(15 : 85→60 : 40, v/v)→MeOH] and HPLC [MeOH–H2O (15 : 85—40 : 60,
v/v), or MeOH–CH3CN–H2O (12 : 8 : 80, v/v/v)] to give everlastosides
K (6, 82.0 mg, 0.020%) and M (8, 369.0 mg, 0.093%) and 10 (19.4 mg,
0.0048%) together with arenariumosides I (18.0 mg, 0.0045%) and II
(15.4 mg, 0.0038%), (2S)-helichrysin (4.0 mg, 0.0010%), apigenin 7-O-
gentiobioside (16.0 mg, 0.0040%), 6-hydroxyluteolin 7-O-b-D-glucopyrano-
side (76.0 mg, 0.019%), kaempferol 3-O-b-D-glucopyranoside (4.8 mg,
0.0012%), kaempferol 3-O-gentiobioside (28.0 mg, 0.0070%), rutin
(13.0 mg, 0.0032%), maltol 3-O-b-D-apiofuranosyl-(1→6)-b-D-glucopyra-
noside (70.0 mg, 0.018%), and phenethyl alcohol b-D-glucopyranosyl-
(1→6)-b-D-glucopyranoside (18.0 mg, 0.0045%).17) Fraction 12 (17.10 g)
was subjected to reversed-phase silica gel CC [500 g, MeOH–H2O
(10 : 90→40 : 60, v/v)→MeOH] and HPLC [MeOH–H2O (30 : 70—45 : 55,
v/v)] to give everlastoside L (7, 6.9 mg, 0.0018%).
Acid Hydrolysis of Everlastosides H (3), I (4), J (5), and K (6) A so-
lution of 3 (1.0 mg) in 1.0 M HCl (1.0 ml) was heated at 80 °C for 3 h. After
cooling, the reaction mixture was neutralized with Amberlite IRA-400 (OHꢀ
form) and then the resin was removed by filtration. Removal of the solvent
from the filtrate under reduced pressure, the residue was separated by Sep-
Everlastoside F (1): A white powder, [a]D28 ꢀ42.5° (cꢂ1.90, MeOH).
High-resolution positive-ion FAB-MS: Calcd for C17H28O12Na (MꢁNa)ꢁ
447.1478; Found 447.1469. UV [lmax (log e), MeOH]: 219 (4.42) nm. IR
(KBr, cmꢀ1): 3550, 1718, 1655, 1075. 1H-NMR (500 MHz, CD3OD) d: