Tannins, flavonol sulfonates, and a norlignan from Phyllanthus virgatus

Article abstract of DOI:10.1021/np970336v

Investigation of the constituents of Phyllanthus virgatus has led to the isolation of five new compounds, including a norlignan, 2-(3,4- methylenedioxybenzyl)-4-(3,4-methylenedioxyphenyl)-3-butyne-1,2-diol named virgatyne (1); a hydrolyzable tannin, virganin (2); and three flavonoid sulfonates, galangin-8-sulfonate (4), galangin-3-O-β-D-glucoside-8- sulfonate (5), and kaempferol-8-sulfonate (6). Their structures were established by spectral and chemical methods.

Full text of DOI:10.1021/np970336v

1194
J . Nat. Prod. 1998, 61, 1194-1197
Ta n n in s, F la von ol Su lfon a tes, a n d a Nor lign a n fr om P h ylla n th u s vir ga tu s
Yu-Lin Huang,†,‡ Chien-Chih Chen,‡ Feng-Lin Hsu,*,† and Chieh-Fu Chen‡
Graduate Institute of Pharmaceutical Sciences, Taipei Medical College, No. 250, Wu-Hsing St., Taipei, Taiwan,
Republic of China, and National Research Institute of Chinese Medicine, No. 155-1, Sec. 2, Li Nung St. Peitou,
Taipei, Taiwan, Republic of China
Received J uly 16, 1997
Investigation of the constituents of Phyllanthus virgatus has led to the isolation of five new compounds,
including a norlignan, 2-(3,4-methylenedioxybenzyl)-4-(3,4-methylenedioxyphenyl)-3-butyne-1,2-diol named
virgatyne (1); a hydrolyzable tannin, virganin (2); and three flavonoid sulfonates, galangin-8-sulfonate
(4), galangin-3-O-â-D-glucoside-8-sulfonate (5), and kaempferol-8-sulfonate (6). Their structures were
established by spectral and chemical methods.
Phyllanthus virgatus Forst. f. (Euphorbiaceae) is an
annual plant in Taiwan, and it is traditionally used to
protect the liver.1 In previous work on P. virgatus, several
lignans and indole-3-carboxylic acid were identified from
the whole plant.2 Continuing our investigation on P.
virgatus, we obtained five additional new compounds
including a norlignan, 2-(3,4-methylenedioxybenzyl)-4-(3,4-
methylenedioxyphenyl)-3-butyne-1,2-diol (virgatyne, 1); a
hydrolyzable tannin (virganin, 2); and three flavonoid
sulfonates, galangin-8-sulfonate (4), galangin-3-O-â-D-glu-
coside-8-sulfonate (5), and kaempferol-8-sulfonate (6).
Twenty-four known compoundss(-)-lirioresinol-B;3 glochi-
done;4 indole-3-carboxaldehyde;5 kaempferol; quercetin;
quercitrin; astragalin; isoquercitrin; rutin; myricitrin; 5;7-
dihydroxy-4′-methoxyflavonol; quercetin-3-O-â-D-glucosyl-
(1f6)-â-D-glucoside; gallic acid; methyl gallate; corilagin;6
1-O-galloyl-â-D-glucose; 1,6-di-O-galloyl-â-D-glucose;7 1,4,6-
tri-O-galloyl-â-D-glucose;8 1,3,4,6-tetra-O-galloyl-â-D-glu-
cose;9 geraniin;10 furosin;11 brevifolin;12 methyl brevifolin-
carboxylate;13 and potassium brevifolincarboxylate14swere
also recognized. The known compounds were identified by
comparison of their spectral data with those in the litera-
ture or by comparison with authentic samples. The
structures of new compounds 1, 2, and 4-6 were estab-
lished by spectral and chemical methods, and the details
are presented herein.
Resu lts a n d Discu ssion
Compound 1 was obtained as white needles, mp 123-
125 °C. The UV spectrum showed absorption bands at λmax
294, 265, and 228 nm. The IR spectrum exhibited bands
at 3322 (OH), 2236 (CtC), 1606 and 1496 (aromatic rings),
and 932 (O-CH2-O) cm-1. The HRMS of 1 displayed a
molecular ion peak at m/z 340.0950 corresponding to
C
19H16O6. The 13C NMR and DEPT-NMR spectra of 1
showed 19 carbons, including four methylene, six methine,
and nine quaternary carbons. In the 1H NMR and 1H-
13C COSY spectra of 1, six aromatic proton signals between
δ 6.44 and 6.61 could be analyzed into a pair of ABX
patterns. The rest of the signals were assigned to two 7′
benzylic protons at δ 2.69 (1H, d, J ) 13.4 Hz) and 2.75
(1H, d, J ) 13.4 Hz), two 1-hydroxymethylene protons at
δ 3.33 (2H, m), and two methylenedioxyl groups at δ 5.64
the carbon signals of four methylene groups were at-
tributed to two methylenedioxyl (δ 100.2 and 100.8) groups,
one benzylic carbon (δ 43.3, C-7′), and one CH2O carbon (δ
68.0, C-1). Among the quaternary resonances, the signals
at δ 85.5 (C-4) and 88.1 (C-3) could be directly assigned to
an alkyne group. The structure of 1 was further estab-
lished using HMBC experiments (Figure 1). Two 7′ ben-
zylic methylene protons at δ 2.69 and 2.75 were correlated
with C-1 (δ 68.0), C-3 (δ 88.1), C-2′ (δ 110.7), and C-6′ (δ
123.3). The proton signal at δ 3.33 (H-1) was correlated
1
(2H, s) and 5.68 (2H, s). Consistent with the H NMR data,
* To whom correspondence should be addressed. Tel.: 886-2-27361661
ext. 672. Fax: 886-2-27370903.
† Taipei Medical College.
‡ National Research Institute of Chinese Medicine.
10.1021/np970336v CCC: $15.00
© 1998 American Chemical Society and American Society of Pharmacognosy
Published on Web 08/26/1998

New Compounds from Phyllanthus
J ournal of Natural Products, 1998, Vol. 61, No. 10 1195
F igu r e 2. 1H-13C long-range correlations observed in the HMBC
spectrum of 5.
F igu r e 1. 1H-13C long-range correlations observed in the HMBC
spectrum of 1.
In the 13C NMR spectrum of 5, the C-3 signal appeared ca.
3 ppm upfield and the C-2 signal (δ 155.2) appeared ca. 9
ppm downfield from those of 4 (δ 145.9),16 suggesting that
the location of this sugar moiety was at C-3. It was further
confirmed by the observation that its anomeric proton
signal (δ 5.56) showed a cross-peak with the carbon signal
at δ 134.6 (C-3) in the HMBC spectrum of 5. The
configuration of the anomeric center was determined to be
â on the basis of the coupling constant of the anomeric
proton signal (δ 5.56, 1H, d, J ) 7.6 Hz). In DEPT and
HMBC spectra of 5 (Figure 2), the proton signal at δ 12.51
(chelated 5-OH) showed a cross-peak by two-bond coupling
with the carbon signal at δ 160.8 and also showed cross-
peaks by three-bond coupling with the carbon signals at δ
110.0 (quaternary carbon) and 98.6 (tertiary carbon). The
assignment of C-5 (δ 160.8), C-10 (δ 110.0), and C-6 (δ 98.6)
was thus confirmed. Therefore, the signal at δ 6.19 was
assignable as H-6, which correlated with the C-6 signal at
δ 98.6. Moreover, the H-6 signal showed cross-peaks with
the carbon signals at δ 110.0 (C-10) and δ 104.3, allowing
us to assign the latter to C-8. Acid hydrolysis17 of com-
pounds 4 and 5 yielded 4a , which was identified as
galangin (5,7-dihydroxyflavonol) by comparison of its 13C
NMR spectral data with those reported in the literature.18
The hydrolysate gave a white precipitate on addition of
barium chloride, which also confirmed the presence of the
sulfonate or the sulfate group.19,20 In addition, the sugar
in the hydrolysate of 5 was identified as glucose by Si gel
TLC comparison21 with authentic sample. Comparing the
1H NMR spectra of 4 and 4a , the signal of ring A changed
from a singlet (4, δ 6.16, H-6) into a pair of meta-coupling
doublets (4a , δ 6.17 and 6.42, each 1H, J ) 1.9 Hz),
indicating the substituent at C-8 was a sulfonate group.
This supposition was supported by the presence of the ion
peaks at m/z 371 [M - H]- and m/z 269 [M - Na - SO3]-
in the FABMS spectrum of compound 4. Furthermore, the
C-8 (δ 103.2) signal in the 13C NMR spectrum of 4 appeared
ca. 10 ppm downfield from that (δ 93.6) of 4a . This spectral
characteristic was in agreement with that of disodium
epicatechin-(4â,5′)-disulfonate,22 which showed that the
aromatic carbon carrying the sulfonyl group shifts down-
field ca. 12 ppm compared to its aglycon.23 In general,
sulfation of a phenolic hydroxy group exhibits upfield
displacement (3.5-5.0 ppm) for the ipso carbon.19-20,24
Based on the above data, compound 4 was identified as
galangin-8-sulfonate. Therefore, compound 5 was estab-
lished as galangin-3-O-â-D-glucoside-8-sulfonate.
with C-7′ (δ 43.3) and C-3 (δ 88.1). These HMBC experi-
mental data suggested that C-2 was located between C-3
and C-7′. Moreover, the carbon signal at δ 85.5 (C-4) was
correlated with aromatic proton signals at δ 6.52 (H-2′′)
and 6.61 (H-6′′), indicating that one of alkynyl carbons was
linked to the phenyl group. The presence of two hydroxyl
groups in 1 was supported by acetylation of 1 with acetic
anhydride in pyridine to afford a monoacetate (1a ) and a
diacetate (1b). Thus, the structure of 1 was determined
to be 2-(3,4-methylenedioxybenzyl)-4-(3,4-methylenedioxy-
phenyl)-3-butyne-1,2-diol, which we named virgatyne.
Compound 2 gave a positive color test with ferric chloride
(a dark blue). The 1H NMR spectral features of 2 were
very similar to those of euphormisin M2 (3),15 except for
the absence of an hexahydroxydiphenoyl (HHDP) group
and existence of three upfield-shifted signals at δ 4.93 (glc
H-3), 4.16 (glc H-6), and 4.01 (glc H-6) assigned with the
1
aid of 1H-1H and H-13C COSY. Comparing the 13C NMR
spectrum of 2 with that of 3, only five carbonyl signals at
δ 164.7, 165.2, 171.6, 172.1, and 176.1 were observed in
the former compound, which supported the absence of the
HHDP group in the structure of 2. Additionally, the
FABMS of 2 showed a molecular ion peak at m/z 622
(C26H22O18) corresponding to the loss of an HHDP group
(302 mass units) from 3 (m/z 924, C40H28O26). The glc H-2
and glc H-4 signals resonated at a lower field (δ 5.13 and
4.93, respectively) than other glucose proton signals,
indicating that the hydroxyl groups at glc C-2 and glc C-4
were acylated. The ABXY-type signals at δ 1.88 (1H, dd,
J ) 3.2, 17.0 Hz), 2.15 (1H, dd, J ) 11.0, 17.0 Hz), 3.63
(dd, J ) 2.9, 11.0 Hz), and 4.93 (overlapped with glc H-3,
4) attributed to two methylene (H-4′) and two methine (H-
3′ and H-2′) protons, respectively, and an aromatic proton
signal at δ 7.26 (1H, s) were due to the acyl moiety at O-2/
O-4 of the glucose core. The glucopyranose ring in 2 was
indicated from the coupling pattern of the sugar proton
1
signals to adopt the C4-conformation.15 Accordingly, this
compound has the structure represented by 2 and was
named virganin.
Compounds 4-6 were characterized as flavonol-8-sul-
fonates on the basis of spectral analyses and chemical
evidence. Compound 4 was obtained as a yellow powder.
Its UV spectrum showed absorption bands at λmax 355, 316,
268, and 213 nm. The negative FABMS of 4 displayed an
ion peak at m/z 371 [M - H]- corresponding to the formula
C15H9O8SNa. In addition, peaks at m/z 349 [M - Na]- and
269 [M - Na - SO3]- suggested the presence of a sodium
sulfate or sodium sulfonate in the structure of 4, and Na+
was also confirmed by atomic absorption spectroscopy. The
aryl proton signal of the A-ring of compound 4 was observed
at δ 6.19 (1H, s), and 13C NMR spectral data indicated that
compound 4 was a flavonol with a 5,6,7- or 5,7,8-trisub-
stituted structure. The 1H and 13C NMR spectra of 5 were
similar to those of 4 except for the presence of signals of a
sugar residue, which indicated that compound 5 was a
glycoside of 4. This observation was consistent with the
negative FABMS spectrum of 5, which displayed an ion
peak at m/z 533 [M - H]-, 163 units higher than that of 4.
1
The H and 13C NMR spectra of compound 6 indicated
that its structure differed from that of compound 4 by the
presence of a hydroxy substitute on the B-ring, in agree-
ment with a molecular ion at m/z 387 [M - H]- (16 units
larger than 4) and [M - Na]- peaks at m/z 365 and m/z
285 [M - Na - SO3]- in the negative FABMS spectrum.
Moreover, acid hydrolysis of compound 6 gave kaempferol,
identified by direct comparison (co-TLC, 1H NMR, and 13C
NMR spectra) with an authentic sample. Accordingly,
compound 6 was determined to be kaempferol-8-sulfonate.
Although flavonoid sulfates and C-O-SO3- linkage, are
known to be present in a number of plant families,19,20 only
a relative few sulfonates such as sodium quercetin-5′-

1196 J ournal of Natural Products, 1998, Vol. 61, No. 10
Huang et al.
Hz, H-6′′), 6.65 (1H, d, J ) 1.6 Hz, H-2′); 13C NMR (CDCl3, 75
MHz) δ 43.3 (C-7′), 68.0 (C-1), 71.5 (C-2), 85.5 (C-4), 88.1 (C-
3), 100.2 and 100.8 (O-CH2-O), 107.1, (C-5′′), 107.8 (C-5′),
110.7 (C-2′), 111.0 (C-2′′), 115.4 (C-1′), 123.3 (C-6′), 125.7 (C-
6′′),129.7 (C-1′′), 146.0 (C-4′), 146.7 (C-3′′), 146.9 (C-3′), 147.5
(C-4′′).
sulfonate, sodium apigenin-3′-sulfonate,25,26 and sodium
procyanidin sulfonate22 have been characterized. These
three compounds (4-6) represent the first known examples
of flavonoid-8-sulfonates from a natural source.
Exp er im en ta l Section
Acetyla tion of Vir ga tyn e (1). Virgatyne (1, 5 mg) in
pyridine (0.5 mL) and Ac2O (1 mL) was allowed to stand at
room-temperature overnight. Usual workup and purification
by preparative TLC (n-hexane-EtOAc 5:1) afforded monoac-
etate 1a (1.5 mg) and diacetate 1b (3.0 mg). Compound 1a :
1H NMR (CDCl3, 200 MHz) δ 2.15 (3H, s, OCOMe-1), 3.00 (2H,
br s, H2-7′), 4.13 (1H, d, J ) 11.2 Hz, H-1), 4.36 (1H, d, J )
11.2 Hz, H-1), 5.96 (2H, s, O-CH2-O), 5.98 (2H, s, O-CH2-
O), 6.73-6.91 (6H, m, aromatic protons). Compound 1b: 1H
NMR (CDCl3, 200 MHz) δ 2.05 (3H, s, OCOMe-2), 2.14 (3H, s,
OCOMe-1), 3.14 (1H, d, J ) 13.6 Hz, H-7′), 3.34 (1H, d, J )
13.6 Hz, H-7′), 4.21 (1H, d, J ) 11.5 Hz, H-1), 4.61 (1H, d, J
) 11.5 Hz, H-1), 5.93 (2H, s, O-CH2-O), 5.95 (2H, s, O-CH2-
O), 6.69-6.94 (6H, m, aromatic protons).
Gen er a l Exp er im en ta l P r oced u r es. IR spectra were
obtained on a Bio-Rad FTS-7 spectrometer. Optical rotations
were measured on a J ASCO DIP-140 digital polarimeter. UV
spectra were recorded on a J ASCO model 7800 UV/vis spec-
trometer. Atomic absorption analysis was performed by a
Hitachi 180-30 atomic absorption spectrometer. EIMS and
FABMS spectra were obtained using a J EOL J MS-D100 and
a J EOL SX-102A spectrometer, respectively. 1H NMR and 13C
NMR spectra were measured with a Bruker AM-300 spec-
trometer and a Varian Gemini-200 spectrometer.
P la n t Ma ter ia l. The whole plants of P. virgatus were
collected at Pintoung, Taiwan, in J uly 1993. A voucher
specimen was deposited in the National Research Institute of
Chinese Medicine.
Vir ga n in (2): brown amorphous powder; [R]25D -43° (c 0.5,
MeOH); UV (MeOH) λmax (log ꢀ) 279 (4.38), 220 (4.66) nm;
HRFABMS m/z 622.0845 [M]-, calcd for C26H22O18, 622.0806;
1H NMR (Me2CO-d6 + D2O, 200 MHz) δ 1.88 (1H, dd, J ) 3.2,
17.0 Hz, H-4′), 2.15 (1H, dd, J ) 11.0, 17.0 Hz, H-4′), 3.63 (1H,
dd, J ) 2.9, 11.0 Hz, H-3′), 4.01 (1H, dd, J ) 6.1, 11.2 Hz, glc
H-6), 4.16 (1H, dd, J ) 6.7, 11.2 Hz, glc H-6), 4.36 (1H, br t,
glc H-5), 4.93 (3H, m, H-2′, glc H-3, 4), 5.13 (1H, dd, J ) 2.2,
3.8 Hz, glc H-2), 6.25 (1H, d, J ) 2.2 Hz, glc H-1), 7.15 (2H, s,
galloyl-H), 7.26 (1H, s, H-3); 13C NMR (Me2CO-d6 + D2O, 50
MHz) δ 30.6 (C-4′), 45.2 (C-3′), 47.8 (C-2′), 60.5 (glc C-3), 62.9
(glc C-6), 72.2 (glc C-4), 72.7 (glc C-2), 78.8 (glc C-5), 92.5 (glc
C-1), 110.1 (galloyl C-2, 6), 114.1 (C-3), 117.5, 118.4, 120.6,
135.9, 139.4, 146.1 (2 × C), 147.7, 164.7, 165.2, 171.6, 172.1,
176.1.
Extr a ction a n d Isola tion . In a previous paper,2 it was
reported that an EtOH extract of the dried whole plants of P.
virgatus (3.0 kg) was chromatographed on a Si gel column
eluting with gradient solvent systems of n-hexane-EtOAc
(10:1 to 0:1) and CH2Cl2-MeOH (10:1 to 0:1) to yield 38
fractions. Fractions 27-30 belonged to the portion eluted with
EtOAc. From fraction 27, indole-3-carboxylic acid was iso-
lated.2 Further separation of fraction 27 through a Si gel
column eluted with CH2Cl2-MeOH (15:1) gave mixtures A and
B. Mixture A was further separated by Sephadex LH-20
(MeOH-H2O 3:1) and preparative silica-TLC (CH2Cl2-Me2-
CO 10:1) to afford virgatyne (1, 34 mg), indole-3-carboxalde-
hyde (17 mg), and (-)-lirioresinol-B (12 mg). Mixture B was
separated by preparative silica-TLC (CH2Cl2-MeOH 6:1) to
give methyl gallate (13 mg). Fractions 28 and 30 were
repeatedly rechromatographed over Sephadex LH-20, eluting
with MeOH, to afford gallic acid (18 mg) from fraction 28, as
well as methyl brevifolincarboxylate (35 mg) and quercitrin
(17 mg) from fraction 30. Fractions 31 and 32 were purified
by elution from Sephadex LH-20 with aqueous MeOH (50%,
75%, 90%). The 75% MeOH eluate was further purified by
GPC on Sephadex LH-20 (MeOH-H2O 3:1) and MPLC (Cos-
mosil C-18, 0-25% aqueous MeOH) to obtain isoquercitrin (33
mg), astragalin (26 mg), 1,6-di-O-galloyl-â-D-glucose (18 mg),
corilagin (47 mg), myricitrin (16 mg), and furosin (7 mg).
Similarly, the 90% MeOH eluate gave virganin (2, 5 mg), 1,4,6-
tri-O-galloyl-â-D-glucose (17 mg), and 1,3,4,6-tetra-O-galloyl-
â-D-glucose (19 mg). Fraction 34 was subjected to the same
procedure as fractions 31 and 32. The 50% MeOH eluate was
purified on Sephadex LH-20 (MeOH-H2O 3:1) to give 1-O-
galloyl-â-D-glucose (12 mg), 5,7-dihydroxy-4′-methoxyflavonol
(25 mg), quercetin-3-â-O-D-glucosyl-(1f6)-â-D-glucoside (32
mg), galangin-3-O-â-D-glucoside-8-sulfonate (5, 42 mg), and
rutin (35 mg). The 75% MeOH eluate was similarly chro-
matographed to afford kaempferol (18 mg), quercetin (15 mg),
galangin-8-sulfonate (4, 37 mg), and kaempferol-8-sulfonate
(6, 25 mg). The 90% MeOH eluate was further purified by
MPLC with aqueous MeOH (10% to 25%) to afford geraniin
(28 mg). Rechromatography of fraction 35 over Sephadex LH-
20 with MeOH-H2O (3:1) furnished potassium brevifolincar-
boxylate (34 mg) and brevifolin (16 mg).
Ga la n gin -8-su lfon a te (4): yellow powder; mp 186-188 °C;
UV (MeOH) λmax (log ꢀ) 213 (4.54), 268 (4.32), 316 (4.12), 355
(4.22) nm; IR (KBr) νmax 3432 (OH), 2918, 1654 (CdO), 1562,
1459, 1424, 1385, 1223 (SdO), 1122 cm-1; negative FABMS
m/z 371 [M - H]- (30), 349 [M - Na]- (100), 269 [M - Na -
SO3]- (17); HRFABMS 348.9987 [M - Na]-, calcd for C15H9O8S,
1
349.0018; H NMR (DMSO-d6, 300 MHz) δ 6.16 (1H, s, H-6),
7.45-7.56 (3H, m, H-3′,4′,5′), 8.50 (2H, dd, J ) 1.9, 8.1 Hz,
H-2′, 6′), 12.50 (1H, br s, 5-OH); 13C NMR (DMSO-d6, 75 MHz)
δ 98.0 (C-6), 103.2 (C-8), 109.9 (C-10), 128.0 (C-2′, 6′), 128.2
(C-3′, 5′), 129.7 (C-4′), 131.0 (C-1′), 137.3 (C-3), 145.9 (C-2),
153.0 (C-9), 160.5 (C-5, 7), 176.1 (C-4).
Acid Hyd r olysis of Ga la n gin -8-su lfon a te (4). Com-
pound 4 (10 mg) in 2 N HCl was heated for 2 h in a boiling
H2O bath to yield galangin (4a ) (2.3 mg), EIMS m/z [M]+ 270
(100). The hydrolysate gave a white precipitate with BaCl2,
which was assumed to be BaSO4.17
Ga la n gin -3-O-â-D-glu cosid e-8-su lfon a te (5): yellow pow-
der; mp 215-217 °C; [R]25 -6° (c 0.6, MeOH); UV (MeOH)
D
λmax (log ꢀ) 216 (4.65), 267 (4.47), 304 (4.24) nm; negative
FABMS m/z 533 [M - H]- (29), 511 [M - Na]- (100), 371 [M
- glucosyl]- (49); 1H NMR (DMSO-d6, 300 MHz) δ 3.09-3.34
(5H, m, glucosyl protons), 3.54 (1H, d, J ) 12.0 Hz, H-6′′), 5.56
(1H, d, J ) 7.6 Hz, H-1′′), 6.19 (1H, s, H-6), 7.49-7.54 (3H, m,
H-3′,4′,5′), 8.49 (2H, dd, J ) 2.0, 7.6 Hz, H-2′, 6′), 12.51 (1H,
s, 5-OH); 13C NMR (DMSO-d6, 75 MHz) δ 60.6 (C-6′′), 69.8,
74.2, 76.4, 77.5 (glucosyl carbons), 98.6 (C-6), 100.8 (C-1′′),
104.3 (C-8), 110.0 (C-10), 128.1 (C-3′, 5′), 129.3 (C-2′, 6′), 130.2
(C-1′), 130.9 (C-4′), 134.6 (C-3), 153.1 (C-9), 155.2 (C-2), 160.8
(C-5), 161.0 (C-7), 177.7 (C-4).
Vir ga tyn e (1): needles from n-hexane; mp 123-125°; [R]25
D
+30° (c 1.0, CH2Cl2); UV (CH2Cl2) λmax (log ꢀ) 294 (3.67), 265
(3.86), 228 (3.98) nm; IR (KBr) νmax 3322 (OH), 2236 (CtC),
1606 and 1496 (aromatic rings), 932 (O-CH2-O) cm-1; EIMS
(70 eV) m/z 340 [M]+ (18), 322 (13), 205 (98), 187 (30), 159
(44), 136 (100), 77 (20); HREIMS 340.0950, calcd for C19H16O6,
340.0947; 1H NMR (CDCl3, 300 MHz) δ 2.32 (1H, br s, -OH),
2.69 (1H, d, J ) 13.4 Hz, H-7′), 2.75 (1H, d, J ) 13.4 Hz, H-7′),
3.19 (1H, br s, -OH), 3.33 (2H, m, H-1), 5.64 (2H, s, O-CH2-
O), 5.68 (2H, s, O-CH2-O), 6.44 (1H, d, J ) 8.1 Hz, H-5′),
6.46 (1H, d, J ) 7.8 Hz, H-5′′), 6.52 (1H, d, J ) 1.6 Hz, H-2′′),
6.56 (1H, dd, J ) 1.6, 8.1 Hz, H-6′), 6.61 (1H, dd, J ) 1.6, 7.8
Acid Hyd r olysis of Ga la n gin -3-O-â-D-glu cosid e-8-su l-
fon a te (5). Compound 5 (10 mg) in 20% H2SO4 was heated
for 1 h in a boiling H2O bath to yield 4a (3.2 mg). After
neutralization, the hydrolysate was analyzed by Si gel TLC
[Kieselgel 60 (Merck Art. 5554), iPrOH-Me2CO-H2O (5:3:
1)].21 It showed a brown spot (Rf 0.46) on TLC over spraying
anilinephthalate solution and heating, which was coincident
with that of glucose.
Ka em p fer ol-8-su lfon a te (6): yellow powder; mp 204-206
°C; UV (MeOH) λmax (log ꢀ) 268 (4.28), 366 (4.41) nm; negative

New Compounds from Phyllanthus
J ournal of Natural Products, 1998, Vol. 61, No. 10 1197
FABMS m/z 387 [M - H]- (40), 365 [M - Na]- (100), 285 [M
- Na - SO3]- (11); HRFABMS 364.9955 [M - Na]-, calcd for
C15H9O9S, 364.9967; 1H NMR (DMSO-d6, 300 MHz) δ 6.13 (1H,
s, H-6), 6.89 (2H, d, J ) 8.8 Hz, H-3′, 5′), 8.37 (2H, d, J ) 8.8
Hz, H-2′, 6′), 12.44 (1H, s, 5-OH); 13C NMR (DMSO-d6, 75 MHz)
δ 97.9 (C-6), 103.0 (C-8), 109.8 (C-10), 115.2 (C-3′, 5′), 121.8
(C-1′), 130.1 (C-2′, 6′), 135.6 (C-3), 147.2 (C-2), 152.6 (C-9),
159.2 (C-4′), 160.3 (C-5), 160.6 (C-7), 175.7 (C-4).
Acid Hyd r olysis of Ka em p fer ol-8-su lfon a te (6). Com-
pound 6 (5 mg) in 2 N HCl was heated for 2 h in a boiling H2O
bath to yield kaempferol (1.6 mg). The hydrolysate gave a
white precipitate with BaCl2, which was assumed to be BaSO4.
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Ack n ow led gm en t. The study was supported by a grant
from the National Science Council of the Republic of China.
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