1224 Short Reports

Viscosic acid (5). Oil (8 mg). UV IzH nm: 215 (108~4.3); REFERENCES
IRvE~‘~~~-~: 2950,2840,26OO(br sh), 1695,1620,1445,1380, 1. Baytop, T (1984) Therapy with Medicinal Plants in Turkey,
1100,1050,980,820,750, ‘H NMR (400 MHz, CDC&) see text; p. 167. Sanal Press, Istanbul.
MS (probe) 70eV (rel. int.): 264 [M]’ (r&Hz403) (methyl ether 2. Gad, S. and Yoel, K. (1970) J. Med. Chem. 13, 1221.
ofviscosicacid)(18),250[M-CH,]+ (16),249[M-Me]+ (68), 3. Suslugas, C., Balansard, G., Julien, J, Gasquet, M., Timon-
233 (13), 203 (20), 156 (31), 117 (98). Davis, P. and Ross, J. C. (1980) Herba Hung. 19, 19; Chem.
Epoxidation of costic acid methyl ester. p-Chloroperbenzoic Abstr. 94, 20290.
acid (10 mg) was added to costic acid methyl ester (5 mg) in 1 ml 4. Herz, W., Hiroaki,C.and Tether, L. H. (1%6)5. Org. Chem. 31,
CHICll in the presence of NaHCOs (10 mg). The mixture was 1632.
left at room temp. for 1 hr. The ‘HNMR spectrum of the
5. Bohhnann, F., Czerson, H. and Schaneweiss, S. (1977)
resulting compound was identical with that of compound 5.
Phytachemistry 11, 2859.
6. Zdero, C., Bohhnann, F., King, R M. and Robinson, H. (1987)
Acknowledgements-This work was supported by the Health Phytochemistry 26, 1207.
Sciences Institute of the University of Istanbul (Grant No. 114). 7. ijksiiz, S. (1976) Pkanta Med. 343.
The authors thank Dr. J. Jakupovic (Berlin) for his kind help in 8. Grande, M., Piera, F., Cuenca, A., Torres, P. and Bellido, I. S.
structure determination. (1985)Planta Med. 414.

Phytochemistry, %A 26, No. 4, pp. 1224-1226, 1987. 003 I -9422/87 $3.00 + 0.00
Printedin GreatBritain. Pergamon Journals Ltd.

A FLAVONOL GLYCOSIDE FROM LYSIMACHlA A4AURlTIANA

KEN YASUKAWA and MICHIO TAKIDO

Department of Pharmacy, College of Science and Technology, Nihon University, Tokyo 101, Japan

(Revised receioed 29 September 1986)

Key Word Index-Lysimachia mauritiana; Primulaceae; mauritianin; kaempferol-3-O-(2,6i-O-a-rhamno-

pyranosyl+galactopyranoside); flavonol glycoside.

Abstract-From the whole plant of Lysimachiu mauritiana, a new flavonol glycoside (mauritianin) was isolated
together with hyperin, kaempferol-3-0-robinobioside and kaempferol-3-0-a-rhamnopyranosyl-(l-2)-8-
galactopyranoside. The structure of mauritianin was established as kaempferol-3-O-(2,6-di-O-a-rhamnopyranosyl-fl-
galactopyranoside).

INTRODUCTION RESULTSAND DISCUSSH)N

Flavor101 glycosides of the kaempferol, quercetin and The concentrated methanol extract prepared from the
myricetin type have already been isolated from the genus air dried plant was extracted successively with ethyl
Lysimachia, a member of the Primulaceae (L. uulgaris acetate and n-butanol. One flavonol glycoside (com-
[l, 23, L. punctata [3] and L. nummuluria [4]). Initial pound 1) was isolated on a Sephadex LH-20column using
chemical investigations of L. mauritiana led to the iso- water-methanol from the ethyl acetate fraction, and
’ lation and structure elucidation of sapogenins [S, 63. In another three tlavonol glycosides (compounds 2-4) were
this paper we report the isolation and characterization of isolated by the same method from the n-butanol fraction.
four flavonol glycosides from this plant. Hyperin and Compound 1, yellow needles, mp 238%239”, gave
kaempferol-3-0-rhamnosyl galactosides are reported for positive ferric chloride and Mg+ HCl tests and was
the first time in Lysimachia; mauritianin is a new com- identified as hyperin by direct comparison with an
pound, which an anti-tumour promoter [7]. authentic sample.
 Short Reports 1225

Compound 2, pale yellow needles, mp 198-200”, also EXPERlMENTAL

gave positive ferric chloride and Mg + HCl tests. The acid
hydrolysis products of 2 showed the presence of kaemp- All mps are uncorr. ‘Hand “C NMR spectra were recorded at
ferol and the two sugars, galactose and rhamnose, by 100 and 25.5 MHz, respectively,chemical shifts are given in
TLC. 2 was subsequently identi6ed as kaempferol-3-O- S(ppm) with TMS as int. std. CC was carried out using
robinobioside by direct comparison with an authentic Sephadex LH-20 (Pharmacia). TLC on Avicel SF (Funakoshi)
sample. was performed with n-BuOH-HOAc-Hz0 (3: 1: 1).
The similarity in the chromophore of compound 3 to Plant material. L. mauritiana Lam. was collected at Miura
that of compound 2 was established by comparison of beach, Kanagawa, Japan in the autumn of 1983.
their colour reactions, acid hydrolysis products and UV Extraction and isolation. Dried whole plant (1.5 kg) was
spectrum. “C NMR also showed that 3 had galactose and extracted with Me-OH and coned to a dark residue, which was
rhamnose units in its structure, the only significant then macerated with hot Hz0 and filtered. The Hz0 soln was
difference from 1 being an upfield shift of 4.1 ppm for the extracted with EtOAc followed by n-BuOH. The EtOAc extract
C-2” of galactose and a downfield shift of 3.5 ppm for the (2g) was then subjected to CC on Sephadex LH-20, using
C-l” of galactose (Table 1). These shifts are analogous to H,G-MeOH as eluant, to yield compound 1 (10 mg). The n-
those reported [S] when the C-2” of galactose is rhamno- BuOH extract (10 g) was analysed by the same method, to yield
sylated in a flavonol glycoside. The structure of com- compounds 2 (10 mg), 3 (20 mg) and 4 (102 mg).
pound 3 was therefore determined as kaempferol-3-O-a- Hyperin. Recrystallization (H,C&MeCN) gave yellow
rhamnopyranosyl-( 1-2)+galactopyranoside. needles, mp 238.5-239”. Analysis: calat: C21H2001Z.H20:
Compound 4, pale yellow needles, mp 202-204”, again C, 52.33; H, 4.76; found: C, 52.29; H, 4.60. Dark green
gave positive ferric chloride and Mg + HCl tests. The acid &our after FeC& reaction, pale red colour in Mg + HCl test.
hydrolysis products of 4 showed the presence of kaemp- UVIzH nm: 258,270sh,293sh, 364. IR v=cm-‘: 3300 (OH),
feral, galactose and rhamnose on TLC. Kaempferol was 2900 (C-H), 1655 (GO), 1605 (C=C), 1060 (C-O). ‘HNMR
identified by direct comparison with an authentic sample. (DMSO-d,,) Gppm: 12.65 (lH, s, OH-5b7.60 (2H, dd, J = 2.0 Hz,
The “CNMR of 4 in DMSO+ also confirmed that it .I = 8.3 Hz, H-6’), 7.59 (lH, d, J = 2.0 Hz, H-2’), 6.86 (lH, d, J
was a glycoside of kaempferol. The “C NMR shifts of the = 8.3 Hz, H-5’). 6.42 (lH, d, J = 1.9 Hz, H-8). 6.21 (IH, d, J
aglycone part of 4 corresponded well to the shifts for = 1.9 Hz, H-6), 5.52 (IH, d, J = 7.3 HZ galactosyl H-l).
kaempferol, the only significant difference being an 13CNMR (DMSO-d,, 90”) Gppm: 177.6 (CA), 164.2 (C-7), 161.3
upfield shift of 2.4 ppm for the C-3. This shift is analogous (C-5), 156.5 (C-2, C-9), 148.4 (C-4’), 144.8 (C-3’), 133.8 (C-3),
to that reported [8] when the 3-hydroxy group is 121.9 (C-6’), 121.4 (C-l’), 116.4 (C-5’), 115.3 (C-2’), 104.2 (C-lo),
glycosylated in a lIavono1 glycoside. UV shifts on the 98.8 (C-6), 93.6 (C-8). For chemical shifts of sugar units see
addition of sodium acetate and aluminium chloride also Table 1.
showed the position of linkage between sugar and ag- Kmnpferol-3-0-robinobioside. Recrystallization (Hz@
lycone as C-3. The “CNMR of 4 is similar to that of MeCN) gave pale yellow needles, mp 198-200”. Analysis: calcd:
compounds 1-3, the only significant difference being an CL7H30016 .3/2HZO: C.50.80; H,5.17; found; C,50.87; H,5.22.
upfield shift of 4.1 and 5.4 ppm, respectively, for the C-2” Dark green colour after FeCIJ reaction, pink colour in Mg + HCI
and C-6” of galactose because it is a rhamnosylated test. UVIEHnm: 263, 3Olsh, 349. IRvEcm-‘: 3400 (OH),
flavonol glycoside [9]. Thus, the structure of compound 4 2950 (C-H), 1660 (GO), 1605 (c--c), 1060 (C-O). ‘HNMR
(named mauritianin) was determined as kaempferol-3-0- (DMSOd6) Gppm: 12.58 (IH, s, OH-5), 8.00 (2H, d, .I = 8.3 Hz,
(2,6di-O-a-rhamnopyranosyl-B-gahctopyranoside). The H-2’, H-6’), 6.89 (ZH, d, .I = 8.3 Hz, H-3’, H-5’), 6.40 (IH, d, J
same trisaccharide has already been found in another = 1.9 Hz, H-S), 6.23 (lH, d, J = 1.9 Hz, H-6), 5.56 (lH, d, J
kaempferol glycoside isolated from Rhamnus nakahami = 7.3 Hz, galactosyl H-l), 5.05 (lH, s, rhamnosyl H-l), 1.05 (3H,
but in this case the sugar is attached to the 4’-OH [9]. d, J = 5.4 Hz, rhamnosyl Me). 13CNMR (DMSO-d,, 90”) 6

Table 1. 13CNMR spectral data of sugar units of compounds 14 (25.5 MHz,

DMSO-d,, TMS as internal standard)

1 2 3 4

GalactoseC-l” 102.6 102.2 99.1 99.2 -

C-2” 71.5 71.5 75.6 75.6 -
C-3” 73.6 13.4 74.1 73.7 -
C-4” 68.2 68.4 68.2 68.3 -
C-5” 75.9 73.9 75.9 74.0 -
C-6” 60.4 65.9 60.4 65.8 -
Rhamnose C-l - loo.4 loo.7 loo.7 loo.3
c-2 - 71.0 70.9 70.9 70.9
c-3 - 70.3 70.8 70.8 10.5
C-4 - 72.6 72.3 72.3 72.3
c-5 - 68.4 68.7 68.8 68.3
C-6 - 17.6 17.2 17.2 17.7
1226 Short Reports

ppm: 177.5 (C-4), 164.3 (C-7), 161.3 (C-S), 160.0 (C-4’), 156.6 (C-2, *H NMR (DMSOC) Gppm: 12.70 (IH, s, OH-5), 8.05 (2H, d, J
C-9), 133.8 (C-3), 130.9 (C-2’, C-6’), 121.1 (C-l’), 115.4 (C-3’,C-S), = 8.3 Hz, H-2’, H-6’), 6.88 (2H, d, J = 8.3 Hz, H-3’, H-5’), 6.41
104.3 (C-lo), 99.0 (C-6), 93.9 (C-8). Chemical shifts of sugar units (IH, d, J = 1.9 Hz, H-6), 6.21 (lH, d, J = 1.9 Hz, H-8), 5.56 (lH,
are given in Table 1. d, J = 7.3 Hz, galactosyl H-l), 5.05 (ZH, s, rhamnosyl H-l, H-l’),
Hydrolysis of2. Compound 2 (3 mg) treated with 2 N HCl at 1.05 (3H, d, J = 5.4 Hz, rhamnosyl Me), 0.79 (3H, d, J = 5.4 Hz,
100” for 3 hr gave kaempferol, galactose and rhamnose. rhamnosyl Me’). 13CNMR (DMSOd6, 90”) dppm: 177.5 (C-4),
Kaempferol was identified by.direct comparison with an authen- 164.0 (C-7), 161.3 (C-S), 159.7 (C-4’), 156.5 (C-2, C-9), 132.9 (C-3),
tic sample. The sugars were identified by TLC with authentic 130.6 (C-2’, C-6’), 121.3 (C-l’), 115.2 (C-3’, C-S), 104.2 (C-lo),
sampies. R, = 0.44 and 0.30 (rhamnose and galactose R, = 0.44 98.9 (C-6), 93.8 (C-8). Chemical shifts of the sugar units are given
and 0.30). in Table 1.
Kaempfeol-3-O-a-rhamnopyranosyl-(l-2)+galac~opyrano- Hydrolysis of 4. Compound 4 (5 mg) treated with 2 N HCI at
side. Recrystallization (H&-MeCN) gave pale yellow needles, 100” for 3 hr gave an aglycone and sugars. The aglycone was
mp 202-204”. Analysis: calcd: C2,HJOOLL.9/5H20: C, 50.73; H, identified as kaempferol by direct comparison with an authentic
5.57; found: C, 5O.io.44,H, 5.27. Dark green colour with Fe-Cl, sample. The sugars were identified as rhamnose and galactose by
reaction, pink colour in Mg + HCl test. UV n%H nm: 262, TLC.
3OOsh, 350. 1 sH+A’CIa mn: 272, 300, 32Osh. 345, 395.
AetoH+*oAcmn: 263, 3OOsh, 349. IRvEcm-‘: 3400 (OH), Acknowledgements-We are grateful to Professor T. J. Mabry,
295 (C-D), 1660 (C==O), 1610 (C==== 1060 (C-D). ‘HNMR Department of Botany, University of Texas at Austin, for a
(DMSO-d,) 6ppm: 12.55 (lH, s, OH-5), 8.07 (2H, d, J = 8.3 HZ, generous gift of kaempferol-3-O-robinobioside. We thank
H-2’, C-6’), 6.85 (2H. d, J = 8.3 Hz, H-3’, H-S), 6.39 (H-I, d, J Y. Kimura, Department of Pharmacy, Nihon University, for IR
= 1.9 Hz, H-8), 6.17 (lH, d, J = 1.9 I-Iq H-6), 5.65 (lH, d, J and Dr. T. Takido, Analytical Centre, College of Science and
= 7.3 Hz, galactosyl H-l), 5.04 (lH, s, rhamnosyl H-l), 0.73 (3H, Technology, Nihon University, for NMR spectra.
d. J = 5.4 Hz, rhamnosyl Me). 1‘C NMR (DMSO-de, 9O”)appm:
177.5 (C-4), 164.8 (C-7), 159.8 (C-4’), 156.5 (C-2), 156.0 (C-9),
REFERENCES
132.9 (C-3), 130.6 (C-2’, C-6’), 121.2 (C-l’), 115.1 (C-3’, C-S),
104.0 (C-10). 98.9 (C-6), 93.7 (C-8). Chemical shifts of sugar units 1. Rzadkowska-Bodalska, H. and Olechnowicz-Stepien, W.
are given in Table 1. (1975) Pol. J. Pharmacol. Pharm. 27, 345.
Hydrolysis of 3. Compound 3 (5 mg) treated with 2 N HCI at 2. Prum, N., Pichon, P. and Raynaud, J. (1972) Plant. Med.
100” for 3 hr gave kaempferol, gaalactose and rhamnose. Phytother. 6,267.
Kaempferol was identified by direct comparison with an authen- 3. Mendez, J. (1970) Experientia 26, 108.
tic sample. The sugars were identified as rhamnose and galactose 4. Popov, V. 1. (1967) Tr. Leningrad. Khim.-Farm. Inst. 21,221.
by TLC. 5. Usmanghani, K. (1977) Pak. J. Sci. Ind. Res. 20, 393.
Kaempfmol-3-0-(2,6di-Os-rhamnopyranosyl-~-galac~opyra- 6. Kitagawa, I., Matsuda, A. and Yosioka, I. (1972)Chem. Pharm.
noside) (mauritianin). Recrystallization (H,O-MeCN) gave pale Bull. 20, 2226.
yellow needles, mp 202-204’. A~lpis: C&d: 7. Yasukawa, K., Takeuchi, M., Sate, Y., Nitta, K. and Takido,
C~,H4~0,9.9/2HzO: C.48.14; H,5.71; found: C,48.23; H,6.01. M. (1985) Proc. Jpn. Cancer Assoc., 44th Meet. 66.
Dark green colour with FeCls reaction, pink colour in Mg + HCI 8. Markham, K. R., Ternai, B., Stanley, R., Geiger, H. and Mabry,
test. UVIzHnm: 264, 3OOsh,348. AEH+*Q nm: 274, 301, T. J. (1978) Tetrahedron 34, 1389.
343, 393.1&?H+N@Acnm: 265, 300, 356. IRvEcm-‘: 3400 9. Lin, C. N., Arisawa, M., Shimizu, M. and Morita, N. (1982)
(OH), 2950 (C-H), 1660 (GO), 1610 (c--c), 1050 (C-D). Phytochemistry 21, 1466.