Flavonoid glycosides from flowers of Sisymbrium officinale and Diplotaxis muralis growing in Georgia

Full text of DOI:10.1007/s10600-012-0234-0

Chemistry of Natural Compounds, Vol. 48, No. 2, May, 2012 [Russian original No. 2, March-April, 2012]
FLAVONOID GLYCOSIDES FROM FLOWERS OF Sisymbrium
officinale AND Diplotaxis muralis GROWING IN GEORGIA
*
M. D. Alaniya, N. Sh. Kavtaradze, A. V. Skhirtladze,
UDC 547.972
M. G. Sutiashvili, and E. P. Kemertelidze
Sisymbrium officinale Scop. (Brassicaceae, Cruciferae), which is indigenous to Georgia, flowers profusely and bears
fruit. The flowers are rich in flavonoids.
Yellow needle-like crystals of 1 (2.1%) formed in the aqueous layer after the appropriate work up [1] of the alcohol
(
80%) extract of the plant flowers and extraction by EtOAc.
The mother liquor remaining after removal of the crystals was fractionated over a polyamide column. The resulting
enriched fraction (6.9% of the starting material) was separated by column chromatography over silica gel (2 ꢀ 65 cm) with
elution by CHCl :EtOH with increasing concentration of the latter. This eluted another two compounds of flavonoid nature,
3
2
and 3.
Compound 1, C H O , MW 610.33 (mass spectrometry), mp 189–192°C, soluble in aqueous EtOH,
2
7 30 16
slightly soluble in concentrated, insoluble in H O, acetone, CHCl . The Bryant reaction [2] was negative. UV spectrum
2
3
(
EtOH, ꢁmax, nm): 360, 340 sh, 250; + CH COONa: 360, 260; + AlCl : 430, 360 sh, 270; + AlCl + HCl: 400, 270.
3 3 3
–
1
IR spectrum (KBr, ꢂmax, cm ): 3425 (OH), 1658, 1604 (ꢃ-pyrone >C=O), 1496 (–CH–).
PMR spectrum (400 MHz, CDCl –CD OD, ꢄ, ppm, J/Hz): 6.48 (1H, d, J = 2.2, H-6), 6.74 (1H, d, J = 2.05, H-8), 7.83
3
3
(
3
(
3
1H, d, J = 2.1, H-2ꢅ), 7.65 (1H, d, J = 8.2, H-6ꢅ), 6.91 (1H, s, H-5ꢅ), 5.04 (1H, d, J = 7.6, H-1ꢅꢅ), 3.82 (1H, br.d, J = 9.4, H-2ꢅꢅ),
.56 (1H, dd, J = 9.4, 3.8, H-3ꢅꢅ), 3.80 (1H, dd, J = 10.0, 9.0, H-4ꢅꢅ), 3.70 (1H, m, H-5ꢅꢅ), 3.54 (1H, dd, J = 12.1, 5.2, H-6ꢅꢅ), 3.72
1H, dd, J = 12.1, 2.2, H-6ꢅꢅ), 5.56 (1H, d, J = 2.0, H-1ꢅꢅꢅ), 4.15 (1H, dd, J = 2.0, 3.0, H-2ꢅꢅꢅ), 3.98 (1H dd, J = 3.1, 10.0, H-3ꢅꢅꢅ),
.8–3.5 (2H, m, H-4ꢅꢅꢅ, H-5ꢅꢅꢅ), 1.22 (3H, d, J = 6, L-rhamnose CH ).
3
1
3
C NMR spectrum [100 MHz, CDCl –CD OD (1:1), ꢄ, ppm]: 156.60 (C-2), 133.20 (C-3), 177.30 (C-4), 161.20
3
3
(
1
C-5), 97.55 (C-6), 162.79 (C-7), 93.01 (C-8), 157.30 (C-9), 105.67 (C-10), 121.10 (C-1ꢅ), 115.20 (C-2ꢅ), 144.70 (C-3ꢅ),
48.65 (C-4ꢅ), 116.45 (C-5ꢅ), 121.62 (C-6ꢅ), 104.6 (C-1ꢅꢅ), 73.60 (C-2ꢅꢅ), 76.12 (C-3ꢅꢅ), 69.35 (C-4ꢅꢅ), 65.80 (C-5ꢅꢅ), 60.82
(
C-6ꢅꢅ), 99.6 (C-1ꢅꢅꢅ), 71.80 (C-2ꢅꢅꢅ), 72.10 (C-3ꢅꢅꢅ), 73.62 (C-4ꢅꢅꢅ), 68.70 (C-5ꢅꢅꢅ), 17.86 (C-6ꢅꢅꢅ).
–
Mass spectrum (EI, 70 eV, m/z, I , %): 609 (100) [M] , 446 (8.6), 463 (2), 301 (9) [3].
rel
Glycoside 1 was hydrolyzed by H SO (2%) to an aglycon of formula C H O , mp 316–318°C, MW 302, quercetin
2
4
15 10 7
[
4]. D-Glucose and L-rhamnose were detected in the carbohydrate part of the hydrolysate.
Alkaline hydrolysis of 1 produced L-rhamnose and a monoside with mp 221–223°C; acid hydrolysis cleaved
D-glucose and quercetin. The monoside was identified as quercetin-3-O-ꢆ-D-glucopyranoside (isoquercitrin) [5, 6].
Enzymatic hydrolysis of 1 by rhamnodiastase gave D-glucose and a monoside with mp 263–266°C that was
characterized as quercetin-7-O-ꢇ-L-rhamnopyranoside [7].
The results of the studies indicated that 1 was identical to quercetin-3-O-ꢆ-D-glucopyranosyl-7-O-ꢇ-L-
rhamnopyranoside [8].
Compound 2 was yellow crystals, MW 756 (mass spectrometry), C H O , mp 219–222°C. The Bryant reaction
3
3 40 20
was negative. UV spectrum (EtOH, ꢁmax, nm): 365, 309 sh, 255; + CH COONa: 360, 285 sh, 260; + CH COONa + H BO :
3
3
3
3
3
60, 295, 260; + AlCl , 400, 303 sh, 260; + AlCl + HCl: 360, 305 sh, 250; + CH ONa: 410, 310 sh, 275.
3 3 3
PMR spectrum (400 MHz, CD OD, ꢄ, ppm, J/Hz): 6.0 (1H, d, J = 2.2, H-6), 6.42 (1H, d, J = 2.1, H-8), 7.90 (1H, d,
3
J = 2.3, H-2ꢅ), 6.91 (1H, d, J = 8.3, H-5ꢅ), 7.63 (1H, dd, J = 2.1, 8.4, H-6ꢅ), 3.81 (3H, s, OCH ), 4.90 (1H, d, J = 7.6, D-galactose
3
H-1ꢅꢅ), 5.62 (1H, d, J = 2.0, L-rhamnose H-1ꢅꢅꢅ), 4.62 (1H, d, J = 7.0, D-xylose H-1ꢅꢅꢅꢅ), 4.0-3.0 (sugar protons), 1.24 (3H, d,
J = 6.0, L-rhamnose CH3).
I. G. Kutateladze Institute of Pharmaceutical Chemistry, 0159, Georgia, Tbilisi, Ul. P. Saradzhishvili 36, fax: (99532) 52 00 23,
e-mail: merialania@yahoo.com. Translated from Khimiya Prirodnykh Soedinenii, No. 2, March–April, 2012, pp. 281–282.
Original article submitted July 25, 2011.
0
009-3130/12/4802-0315 ©2012 Springer Science+Business Media, Inc.
315

1
3
C NMR spectrum [100 MHz, CDCl –CD OD (1:1), ꢄ, ppm]: 157.72 (C-2), 133.10 (C-3), 177.13 (C-4), 161.08
3
3
(
(
C-5), 97.0 (C-6), 161.45 (C-7), 93.02 (C-8), 156.07 (C-9), 105.68 (C-10), 121.10 (C-1ꢅ), 114.02 (C-2ꢅ), 149.40 (C-3ꢅ), 149.55
C-4ꢅ), 115.08 (C-5ꢅ), 122.30 (C-6ꢅ), 55.60 (OCH ), 102.9 (D-galactose C-1ꢅꢅ), 73.72 (C-2ꢅꢅ), 74.75 (C-3ꢅꢅ), 66.92 (C-4ꢅꢅ),
3
7
(
5.80 (C-5ꢅꢅ), 66.80 (C-6ꢅꢅ); 100.2 (L-rhamnose C-1ꢅꢅꢅ), 79.32 (C-2ꢅꢅꢅ), 76.0 (C-3ꢅꢅꢅ), 69.95 (C-4ꢅꢅꢅ), 67.90 (C-5ꢅꢅꢅ), 15.35
C-6ꢅꢅꢅ); 105.60 (D-xylose C-1ꢅꢅꢅꢅ), 75.50 (C-2ꢅꢅꢅꢅ), 77.62 (C-3ꢅꢅꢅꢅ), 72.60 (C-4ꢅꢅꢅꢅ), 65.72 (C-5ꢅꢅꢅꢅ).
Flavonoid 2 was hydrolyzed by H SO4 (2%) to an aglycon with mp 300–304°C, D-xylose, D-galactose, and
2
L-rhamnose. The algycon was identified as 3,5,7,4ꢅ-tetrahydroxy-3ꢅ-methoxyflavone or isorhamnetin [1].
A comparative analysis of UV spectra of the glycoside and the aglycon taken with added diagnostic reagents showed
that the carbohydrates were located on C-3 and C-7.
Enzymatic hydrolysis of 2 by rhamnodiastase gave D-galactose and a bioside; acid hydrolysis cleaved isorhamnetin,
D-xylose, and L-rhamnose. Therefore, the bioside was isorhamnetin-7-O-xylosidorhamnoside. The resonance for ꢇ-L-rhamnose
H-1 appeared in the PMR spectrum at 5.60 ppm, which indicated it was located on C-7 of the aglycon. The chemical shift of
D-xylose H-1 was 4.68 ppm (J = 7.0), which indicated it was bonded to rhamnose at C-2 because L-rhamnose C-2 experienced
1
3
a shift effect (ꢄ 79.32) in the C NMR spectrum. The resonance for D-galactose H-1 at 4.90 ppm (d, J = 7.6) and the result of
the enzymatic hydrolysis indicated that it was bonded to C-3 of the aglycon.
It was concluded from the
results that flavonoid 2 was isorhamnetin-3-O-ꢆ-D-galactopyranosyl-
7
-O-ꢇ-L-rhamnopyranosyl-(2ꢈ1)-ꢆ-D-xylopyranoside.
Compound 3 was yellow needle-like crystals, C H O , MW 742 (mass spectrometry). UV spectrum (EtOH,
3
2 38 20
ꢁmax, nm): 350, 300 sh, 260; + AlCl : 420, 270; + AlCl + HCl: 400, 350 sh, 270; + CH COONa: 350, 260; + CH COONa +
3
3
3
3
–
1
H BO : 370, 260; + CH ONa: 390. IR spectrum (KBr, ꢂ , cm ): 3435 (OH), 1660, 1610 (ꢃ-pyrone >C=O), 1493 (–CH–).
3
3
3
max
PMR spectrum [400 MHz, CDCl –CD OD (1:1), ꢉꢄ, ppm, J/Hz]: 7.80 (1H, d, J = 2.1, H-2ꢅ), 7.65 (1H, d, J = 8.2,
3
3
H-6ꢅ), 6.79 (1H, d, J = 8.2, H-5ꢅ), 6.49 (1H, d, J = 2, H-6), 6.28 (1H, d, J = 2.2, H-8), 5.20 (1H, d, J = 8, D-galactose H-1ꢅꢅ), 5.52
1H, d, J = 2.2, L-rhamnose H-1ꢅꢅꢅ), 4.60 (1H, d, J = 7.0, D-xylose H-1ꢅꢅꢅꢅ), 4.0-3.0 (sugar protons), 1.25 (3H, d, J = 6.1,
L-rhamnose CH ).
(
3
1
3
C NMR spectrum [100 MHz, CDCl –CD OD (1:1), ꢄ, ppm]: 156.60 (C-2), 133.20 (C-3), 177.30 (C-4), 161.20
3
3
(
1
C-5), 97.55 (C-6), 162.79 (C-7), 93.01 (C-8), 157.30 (C-9), 105.67 (C-10), 121.10 (C-1ꢅ), 115.20 (C-2ꢅ), 144.70 (C-3ꢅ),
48.65 (C-4ꢅ), 116.45 (C-5ꢅ), 121.62 (C-6ꢅ), 103.5 (C-1ꢅꢅ), 73.60 (C-2ꢅꢅ), 74.72 (C-3ꢅꢅ), 66.86 (C-4ꢅꢅ), 75.80 (C-5ꢅꢅ), 66.79
(
(
C-6ꢅꢅ); 100.4 (C-1ꢅꢅꢅ), 79.32 (C-2ꢅꢅꢅ), 75.70 (C-3ꢅꢅꢅ), 69.90 (C-4ꢅꢅꢅ), 67.90 (C-5ꢅꢅꢅ), 15.30 (C-6ꢅꢅꢅ); 105.62 (C-4ꢅꢅꢅꢅ), 75.49
C-2ꢅꢅꢅꢅ), 77.63 (C-3ꢅꢅꢅꢅ), 72.55 (C-4ꢅꢅꢅꢅ), 65.56 (C-5ꢅꢅꢅꢅ).
Acid hydrolysis of 3 produced quercetin, D-galactose, D-xylose, and L-rhamnose. A comparison of UV spectra of 3
and its aglycon indicated that the carbohydrates were situated on C-3 and C-7. The products of enzymatic hydrolysis of 3 were
D-galactose and a bioside that was cleaved by acid hydrolysis into quercetin, D-xylose, and L-rhamnose. A comparison of
1
3
PMR and C NMR spectra of the carbohydrate parts of 3 and 2 indicated that they were identical. Therefore, the product of
enzymatic hydrolysis was quercetin-7-O-ꢇ-L-rhamnopyranosyl-(2ꢈ1)-O-ꢆ-D-xylopyranoside.
The results suggested that 3 had the structure quercetin-3-O-ꢆ-D-galactopyranosyl-7-O-ꢇ-L-rhamnopyranosyl-(2ꢈ1)-
O-ꢆ-D-xylopyranoside.
A yellow crystalline precipitate consisting of two compounds that were identical to 2 and 3 after separation was
isolated from flowers of Diplotaxis muralis (L.) DC. by the method described above.
Isorhamnetin-3-O-ꢆ-D-galactopyranosyl-7-O-ꢇ-L-rhamnopyranosyl-(2ꢈ1)-ꢆ-D-xylopyranoside (2) and quercetin-
3
-O-ꢆ-D-galactopyranosyl-7-O-ꢇ-L-rhamnopyranosyl-(2ꢈ1)-O-ꢆ-D-xylopyranoside (3) were not previously reported.
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3