Rhamnetin Glycosides from the Genus Spiraea

Article abstract of DOI:10.1007/s10600-018-2255-9

Rhamnetin (7-O-methylquercetin,1) and its glycosides were found for the first time in the genus Spiraea (Rosaceae) during chromatographic studies of representatives from the subgenus Protospiraea. Leaves of S. salicifolia yielded1, rhamnetin-3-O-β-D-glucopyranoside (2), and two new flavonoids3and4that were identified by UV, IR, and NMR spectroscopy and mass spectrometry as rhamnetin-3-O-(6″-O-p-coumaroyl)-β-D-glucopyranoside (spiraearhamnin A,3) and rhamnetin-3-O-(6″-O-caffeoyl)-β-D-glucopyranoside (spiraearhamnin B,4). Leaves of S. betulifolia and S. betulifolia var. aemiliana afforded1and2and glycosides of kaempferol, quercetin, and isorhamnetin. Species of the subgenus Metaspiraea (S. alpina, S. chamaedryfolia, S. dahurica, S. hypericifolia, and S. media) did not contain1or its derivatives.

Full text of DOI:10.1007/s10600-018-2255-9

DOI 10.1007/s10600-018-2255-9
Chemistry of Natural Compounds, Vol. 54, No. 1, January, 2018
RHAMNETIN GLYCOSIDES FROM THE GENUS Spiraea
1
2
D. N. Olennikov and N. K. Chirikova
Rhamnetin (7-O-methylquercetin, 1) and its glycosides were found for the first time in the genus Spiraea
Rosaceae) during chromatographic studies of representatives from the subgenus Protospiraea. Leaves of
(
S. salicifolia yielded 1, rhamnetin-3-O-ꢀ-D-glucopyranoside (2), and two new flavonoids 3 and 4 that were
identified by UV, IR, and NMR spectroscopy and mass spectrometry as rhamnetin-3-O-(6ꢁ-O-p-coumaroyl)-
ꢀ
-D-glucopyranoside (spiraearhamnin A, 3) and rhamnetin-3-O-(6ꢁ-O-caffeoyl)-ꢀ-D-glucopyranoside
(
spiraearhamnin B, 4). Leaves of S. betulifolia and S. betulifolia var. aemiliana afforded 1 and 2 and
glycosides of kaempferol, quercetin, and isorhamnetin. Species of the subgenus Metaspiraea (S. alpina,
S. chamaedryfolia, S. dahurica, S. hypericifolia, and S. media) did not contain 1 or its derivatives.
Keywords: Spiraea, Rosaceae, rhamnetin-3-O-(6ꢁ-O-p-coumaroyl)-ꢀ-D-glucopyranoside, rhamnetin-3-O-(6ꢁ-O-
caffeoyl)-ꢀ-D-glucopyranoside, spiraearhamnin, Protospiraea, Metaspiraea.
The genus Spiraea (Rosaceae) is an interesting subject for chemotaxonomic studies because of the broad distribution
and various opinions about the number of species, which are complicated by hybridization and polymorphism. Flavonoids of
various structural types are considered marker compounds for this genus and the family as a whole [1]. Characteristic flavonoids
of the genus Spiraea are the flavonols kaempferol and quercetin and their glycosides [1], which were detected in S. canescens
D. Don [2], S. cantoniensis Lour. [3], S. chamaedryfolia L. [4], S. formosana Hayata [5], S. hypericifolia L., S. media Schmidt
[
4], and S. salicifolia L. [6]. Methoxylated quercetin derivatives including rhamnazin and the dimeric flavonols sparin A and
B, which were atypical of the genus, were relatively recently isolated from S. brahuica Boiss. [7]. Isorhamnetin was also
observed in several Siberian Spiraea species [8]. This suggested that methoxylated flavonols were more broadly distributed
within this genus.
Previously, the isolation of kaempferol and quercetin glycosides and their acylated derivatives from S. salicifolia was
reported by us [9]. In continuation of research on Spiraea flavonoids, we detected traces of rhamnetin (7-O-methylquercetin)
in the hydrolysate of leaves from S. salicifolia. Herein, results from studies of several Spiraea species for the presence of
rhamnetin and its derivatives are presented.
Four compounds were isolated by chromatographic separation from the EtOAc fraction of S. salicifolia and were
identified as rhamnetin (1), rhamnetin-3-O-ꢀ-D-glucopyranoside (2), and two new flavonoid glycosides 3 and 4.
OH
3'
OH
O
H
3
CO
7
O
O
5'
R
O
6''
9'''
3
'''
3
O
O
4''' OH
OH
OH
OH
1''
HO
3
, 4
3: R = H; 4: R = OH
1
) Institute of General and Experimental Biology, Siberian Branch, Russian Academy of Sciences, 6 Sakhꢂyanovoi
St., Ulan-Ude, 670047, fax: 7 (3012) 43 47 43, e-mail: olennikovdn@mail.ru; 2) M. K. Ammosov North-Eastern Federal
University, 58 Belinskogo St., Yakutsk, 677000, e-mail: hofnung@mail.ru. Translated from Khimiya Prirodnykh Soedinenii,
No. 1, January–February, 2018, pp. 38–42. Original article submitted June 3, 2017.
0
009-3130/18/5401-0041 ©2018 Springer Science+Business Media, LLC
41

+
Compound 3 had the formula C H O according to HR-ESI-MS (625.218 [M + H] ; calcd 625.591). The UV and
3
1 28 14
IR spectra indicated that 3 was an acylated flavonoid glycoside [10]. The flavonoid aglycon 3a, D-glucose, and p-coumaric
2
acid were identified in its hydrolysis products. The ESI-MS spectrum of 3a showed two strong ions with m/z 317 and 302
+
+
that corresponded to the pseudomolecular ion [M + H] and its demethylated fragment [(M + H) – 15] .
1
3
A comparison of its C NMR spectrum with that of quercetin indicated that C-7 underwent a weak-field shift
(
ꢃ 163.8ꢄ164.8) with C-6 (ꢃ 98.2ꢄ97.3) and C-8 (ꢃ 93.7ꢄ92.5) experiencing simultaneously strong-field shifts [11]. These
features allowed 3a to be characterized as rhamnetin (7-O-methylquercetin). HPLC analyses also confirmed that the retention
times of 3a and rhamnetin standard sample were similar and different from those of isorhamnetin (3ꢂ-O-methylquercetin) and tamarixetin
+
(
4ꢂ-O-methylquercetin). The ESI-MS of 3 gave peaks for a pseudomolecular ion [M + H] with m/z 625 and fragments formed
+
+
by sequential cleavage of p-coumaric acid (479) [(M + H) –146] and glucopyranose (317) [(M + H) – 146 – 162] . This
indicated that the probable structure was a rhamnetin glycoside with an acylated carbohydrate [12, 13].
1
3
The PMR and C NMR spectra were similar to those of rhamnetin-3-O-ꢀ-D-glucopyranoside (2) (Table 1) [14].
Glycosylation of flavonoid C-3 was indicated by shifts of C-3 (ꢃ 136.2ꢄ133.5) and C-2 (ꢃ 147.5ꢄ157.2) as compared with
those of rhamnetin (3a) and correlations in the HMBC spectrum between resonances for glucopyranose H-1ꢁ (ꢃ 5.44) and
rhamnetin C-3 (ꢃ 133.5) [15]. Additional resonances in the PMR spectrum were attributed to protons of a trans double bond
[
ꢃ 6.28 (1H, d, J = 16.0) and 7.40 (1H, d, J = 16.0)] and a para-substituted benzene ring [ꢃ 6.78 (2H, d, J = 9.0) and 7.92 (2H,
d, J = 9.0)] in p-coumaric acid. Resonances of glucopyranose H-6ꢁ (ꢃ 4.45 and 4.63) and C-6ꢁ (ꢃ 64.1) were shifted to weak
field, indicating that an acyl group added to carbohydrate C-6ꢁ. HMBC spectra in which correlations were observed
between resonances of glucopyranose H-6ꢁ (ꢃ 4.45 and 4.63) and p-coumaric acid carbonyl C-9ꢂꢁ (ꢃ 166.5) confirmed this.
The results determined the structure of 3 as rhamnetin-3-O-(6ꢁ-O-p-coumaroyl)-ꢀ-D-glucopyranoside, which was named
spiraearhamnin A.
+
Compound 4 had the formula C H O (HR-ESI-MS m/z 641.402 [M + H] ; calcd 641.591). The hydrolysis
3
1 28 15
products of 4 were rhamnetin, D-glucose, and caffeic acid. The ESI-MS spectrum gave a peak for the protonated ion with
+
m/z 641 [M + H] and peaks for fragments with m/z 479 and 317 due to sequential cleavage of caffeic acid and glucose [3].
1
3
A comparison of the PMR and C NMR spectra showed that they were similar to those of 3 but differed in the positions of
resonances from a 1,3,4-trisubstituted benzene ring including proton resonances at ꢃ 7.12 (H-2ꢂꢁ), 6.80 (H-5ꢂꢁ), and 6.89
1
3
(
[
H-6ꢂꢁ) and C resonances at ꢃ 127.0 (C-1ꢂꢁ), 115.0 (C-2ꢂꢁ), 146.5 (C-3ꢂꢁ), 148.0 (C-4ꢂꢁ), 116.5 (C-5ꢂꢁ), and 123.3 (C-6ꢂꢁ)
16]. Correlations in the HMBC spectrum between resonances of glucopyranose proton H-6ꢁ (ꢃ 4.40 and 4.69) and
caffeic acid C-9ꢂꢁ (ꢃ 166.2) indicated that the latter was bonded to carbohydrate C-6ꢁ. Compound 4 was identified based on
the results as rhamnetin-3-O-(6ꢁ-O-caffeoyl)-ꢀ-D-glucopyranoside, which was named spiraearhamnin B.
Acylated rhamnetin glycosides still represent a rare flavonoid group with only two known members, i.e., 3ꢁ-O-p-
coumaroylxanthorhamnin B [rhamnetin-3-O-(3ꢁꢁ-O-p-coumaroyl)rhamnoside] from Rhamnus petiolaris Boiss. (Rhamnaceae)
[
[
12] and campylospermoside F [rhamnetin-4ꢂ-O-(6ꢁ-O-p-coumaroyl)-ꢀ-D-glucopyranoside] from Ouratea calantha Gilg
Campylospermum calanthum (Gilg) Farron, Ochnaceae] [17].
The occurrence of rhamnetin (1) in the genus Spiraea was determined using chromatography of acid-hydrolyzed
alcohol extracts from seven Spiraea species. Two species that were representatives of the subgenus Protospiraea like
S. salicifolia, i.e., S. betulifolia and S. betulifolia var. aemiliana, contained 1 in addition to kaempferol, quercetin, and
isorhamnetin. The main flavonoid aglycons from species of the subgenus Metaspiraea, i.e., S. alpina, S. chamaedryfolia,
S. dahurica, S. hypericifolia, and S. media, were quercetin, kaempferol, and isorhamnetin. Compound 1 was not observed in
these species. Chromatography of the EtOAc fraction of S. betulifolia detected 2 and flavonol glycosides that were identified
as astragalin (5), nicotiflorin (6), tiliroside (7), isoquercitrin (8), hyperoside (9), avicularin (10), rutin (11), helichrysoside
(
12), quercetin-3-O-(6ꢁ-caffeoyl)-ꢀ-D-glucopyranoside (13), and isorhamnetin-3-O-ꢀ-D-glucopyranoside (14). Runners of
S. betulifolia var. aemiliana yielded 2, 7–9, 11, 12, and 14. The chemistry of S. betulifolia and S. betulifolia var. aemiliana was
studied for the first time.
Thus, the results showed that the genus Spiraea contained kaempferol, quercetin, and isorhamnetin and possibly also
rhamnetin, the distribution of which was probably limited to species of the subgenus Protospiraea.
4
2

TABLE 1. PMR (500 MHz) and 13C NMR Spectra (125 MHz) of 3 and 4 (MeOH-d , ꢃ, ppm, J/Hz)
4
3
4
C atom
ꢃH
ꢃC
ꢃH
ꢃC
Rhamnetin
2
3
4
5
6
7
8
9
–
–
–
–
157.2
133.5
177.0
161.0
97.3
164.8
92.5
156.3
103.9
122.6
116.0
145.0
148.6
115.5
120.4
56.2
–
–
–
–
156.9
133.2
177.2
160.9
97.5
165.0
92.0
156.4
104.2
122.7
115.9
144.8
148.5
115.3
121.0
56.2
6.35 (1Í, d, J = 2.1)
6.40 (1Í, d, J = 2.0)
–
–
6.65 (1Í, d, J = 2.2)
6.62 (1Í, d, J = 2.1)
–
–
–
–
–
–
10
1
2
3
4
5
6
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
7.72 (1Í, d, J = 2.0)
7.75 (1Í, d, J = 2.1)
–
–
–
–
6.93 (1Í, d, J = 9.0)
7.66 (1Í, dd, J = 9.0, 2.0)
3.85 (3H, s)
6.94 (1Í, d, J = 9.1)
7.69 (1Í, dd, J = 9.1, 2.1)
3.81 (3H, s)
7
-OCH3
3-O-ꢀ-D-Glucopyranosyl
1ꢂꢂ
2ꢂꢂ
3ꢂꢂ
4ꢂꢂ
5ꢂꢂ
6ꢂꢂ
5.44 (1Í, d, J = 7.1)
100.5
73.7
76.9
70.1
77.5
64.1
5.37 (1Í, d, J = 7.0)
3.04–3.31 (4H, m)
100.3
73.9
76.5
70.0
77.7
63.8
3.07–3.35 (4H, m)
4.45 (1Í, dd, J = 11.5, 2.3)
.63 (1Í, dd, J = 11.5, 5.3)
4.40 (1Í, dd, J = 11.6, 2.3)
4.69 (1Í, dd, J = 11.6, 5.4)
6ꢂꢂ-O-Caffeoyl
4
6ꢂꢂ-O-p-Coumaroyl
1
2
3
4
5
6
7
8
9
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
ꢂꢂꢂ
–
126.4
129.8
116.8
157.9
116.8
129.8
145.6
114.3
166.5
127.0
115.0
146.5
148.0
116.5
123.3
145.6
114.5
166.2
7.92 (2Í, d, J = 9.0)
6.78 (2Í, d, J = 9.0)
–
6.78 (2Í, d, J = 9.0)
7.92 (2Í, d, J = 9.0)
7.40 (1Í, d, J = 16.0)
6.28 (1Í, d, J = 16.0)
–
7.12 (1Í, d, J = 2.0)
–
–
6.80 (1Í, d, J = 8.0)
6.89 (1Í, dd, J = 8.0, 2.1)
7.45 (1Í, d, J = 16.0)
6.12 (1Í, d, J = 16.0)
–
EXPERIMENTAL
General Comments. Column chromatography (CC) used SiO , reversed-phase SiO (RP-SiO ), polyamide (Sigma-
2
2
2
Aldrich, St. Louis, MO, USA), and Sephadex LH-20 (GE Healthcare, Little Chalfont, UK). IR spectra were recorded from
KBr pellets (1:100) on an FT-801 FTIR spectrometer (NPF Simex, Novosibirsk, Russia). Spectrophotometric studies used an
SF-2000 spectrophotometer (OKB Spektr, St. Petersburg, Russia); mass spectrometric, an LCMS-8050 TQ-mass spectrometer
(
Shimadzu, Columbia, MD, USA) with electrospray ionization (ESI, positive-ion mode), ESI interface temperature 300°C,
desolvation line temperature 250°C, heating block temperature 400°C, spraying gas (N ) flow rate 3 L/min, heating gas (air)
2
flow rate 10 L/min, collision-induced dissociation gas (CID gas, Ar) pressure 270 kPa, Ar flow rate 0.3 mL/min, capillary
potential 3 kV, and mass scan range (m/z) 100–1,000. NMR spectra were recorded on a VXR 500S NMR spectrometer
(
Varian, Palo Alto, CA, USA). Preparative HPLC used a Summit liquid chromatograph (Dionex, Sunnyvale, CA, USA).
Plant Material. Runners of Spiraea species were collected during flowering in various districts of Russia: S. alpina
Pall., in the vicinity of Nilov Desert (Tunkinskii District, Republic of Buryatia; Jul. 21, 2016; 51°51ꢂ15ꢁ N, 101°43ꢂ3ꢁ E,
4
3

h 2,318 m above sea level); S. betulifolia Pall., Nizhnetambovskoe village (Komsomolꢂskii District, Khabarovskii Krai, Jul.
4, 2014; 50°57ꢂ31ꢁ N, 159°56ꢂ15ꢁ E, h 84 m above sea level); S. betulifolia var. aemiliana (C. K. Schneid.) Koidz.
syn. S. beauverdiana C. K. Schneid.), Kozyrevsk town (Ust-Kamchatskii District, Kamchatskii Krai; Jul. 21, 2016; 56°1ꢂ40ꢁ N,
38°8ꢂ26ꢁ E, h 96 m above sea level); S. chamaedryfolia L. (syn. S. flexuosa Fisch. ex Cambess., S. ussuriensis Pojark.),
1
(
1
Boyarsk village (Kabanskii District, Republic of Buryatia; Jun. 12, 2015; 51°50ꢂ57ꢁ N, 106°4ꢂ12ꢁ E, h 458 m above sea level);
S. dahurica (Rupr.) Maxim., Malovskii town (Bauntovskii District, Republic of Buryatia; Jul. 20, 2014; 54°23ꢂ34ꢁ N, 113°33ꢂ15ꢁ E,
h 867 m above sea level); S. hypericifolia L. (syn. S. aquilegiifolia Pall.), in the vicinity of Merkit Fortress (Mukhorshibirskii
District, Republic of Buryatia; Jun. 19, 2016; 51°9ꢂ6ꢁ N, 107°6ꢂ30ꢁ E, h 928 m above sea level); S. media Schmidt
(syn. S. sericea Turcz.), in the vicinity of Sukhoi Ruchei (Kabanskii District, Republic of Buryatia; Jun. 18, 2015; 51°48ꢂ33ꢁ N,
1
2
06°1ꢂ22ꢁ E, h 435 m above sea level); and S. salicifolia L., Kyren village (Tunkinskii District, Republic of Buryatia; Jul. 20,
015; 51°40ꢂ48ꢁ N, 102°11ꢂ54ꢁ E, h 741 m above sea level). The species were determined by Dr. T. A. Aseeva (IGEB, SB,
RAS). Raw material was dried in a convection oven (50°C) to ꢅ5% moisture content.
Isolation of 1–4 from S. salicifolia. The extraction conditions and production of the EtOAc fraction (SF-2) and its
subfractions (SF-2/3-01, SF-2/3-06) were reported earlier [9]. Subfraction SF-2/3-01 was separated over SiO (CC, 1 ꢆ 30 cm)
2
using a hexane–EtOAc gradient (95:5) to isolate rhamnetin (1, 18 mg) [18]. Subfraction SF-2/3-06 was chromatographed
over SiO (CC, 3 ꢆ 30 cm; hexane–EtOAc, 92:8) and then by preparative HPLC [LiChrospher PR-18 column (250 ꢆ 10 mm,
2
#
10 ꢇm,; Supelco, Bellefonte, PA, USA); mobile phase H O (A) and MeCN (B); gradient mode (%B): 0–40 min, 40–60%;
2
4
0–60 min, 60–90%; flow rate ꢈ 1 mL/min; column temperature 30°C, UV detector at 350 nm] to afford rhamnetin-3-O-ꢀ-D-
glucopyranoside (2, 26 mg) [14], 3 (37 mg), and 4 (28 mg).
Spiraearhamnin A (3). Ñ Í Î . HR-ESI-ÌS m/z 625.218 [M + Í] ; calcd 625.591. (+)ESI-ÌÑ m/z: 625
+
3
1 28 14
+
+
+
[
(
M + Í] , 479 [(M + Í) – 146] , 317 [(M + Í) – 146 – 162] . UV spectrum (ÌåÎÍ, ꢉ , nm): 258, 317, 361. IR spectrum
max
13
–
1
ꢈmax, cm ): 3341, 1720, 1641, 1625, 1604. Table 1 presents the PMR (500 MHz) and C NMR spectra (125 MHz).
+
Spiraearhamnin B (4). Ñ Í Î . HR-ESI-ÌS m/z 641.402 ([M + Í] ; calcd 641.591. (+)ESI-ÌÑ m/z: 641
3
1 28 15
+
+
+
[
(
M + Í] , 479 [(M + Í) – 162] , 317 [(M + Í) – 2 ꢆ 162] . UV spectrum (ÌåÎÍ, ꢉ , nm): 256, 334, 368. IR spectrum
ꢈmax, cm ): 3327, 1725, 1637, 1620, 1601. Table 1 presents the PMR (500 MHz) and C NMR spectra (125 MHz).
max
13
–
1
Acid Hydrolysis of 3 and 4. The compound (5 mg) was dissolved in TFA (10 mL, 5%) and heated at 110°C for 2 h.
The hydrolysate was concentrated in vacuo, dissolved in MeOH, and chromatographed over polyamide (CC, 20 g) with
elution successively by H O (100 mL, eluate I) and EtOH (90%, 250 mL, eluate II). The obtained eluates were concentrated
2
in vacuo and analyzed by HPLC (conditions 1, monosaccharides as 3-methyl-1-phenyl-2-pyrazolin-5-one derivatives [19];
conditions 2, phenolic compounds). Eluate I was also analyzed as before to determine if the monosaccharides were the D- or
L-form [20]. Eluate I after hydrolysis of 3 and 4 contained glucose (t 12.50 min); eluate II, rhamnetin (t 14.51 min); in
R
R
addition to p-coumaric acid for 3 (t 8.12 min) and caffeic acid for 4 (t 6.89 min). Eluates II were also separated over SiO
R
R
2
(
CC, 1 ꢆ 15 cm, hexane–EtOAc, 100:0ꢄ60:40) to isolate from 3 rhamnetin (3a, 2 mg) and p-coumaric acid (1 mg) [11] and
from 4, rhamnetin (3a, 2 mg) [18] and caffeic acid (1 mg) [21].
+
+
Rhamnetin (3a). Ñ Í Î . (+)ESI-ÌÑ m/z: 317 [M + Í] , 302 [(M + Í) – 15] . UV spectrum (ÌåÎÍ, ꢉmax, nm):
1
6 12 7
2
1
57, 369. 13C NMR spectrum (125 MHz, ÌåÎÍ-d , ꢃ, ppm): 146.8 (C-2), 136.3 (C-3), 176.0 (C-4), 160.3 (C-5), 97.3 (C-6),
4
64.9 (C-7), 92.0 (C-8), 156.1 (C-9), 104.3 (C-10), 122.5 (C-1ꢂ), 115.8 (C-2ꢂ), 145.0 (C-3ꢂ), 148.4 (C-4ꢂ), 115.4 (C-5ꢂ), 120.3
(C-6ꢂ), 56.1 (7-OCH3).
Acid Hydrolysis of Spiraea Extracts. A weighed portion of plant raw material (0.4 g) was mixed with
hexamethylenetetramine solution (1 mL, 0.5%), Me CO (20 mL), and HCl solution (7 mL, 25%), refluxed for 30 min, and
2
cooled. The extract was filtered, treated with H O (25 mL) and EtOAc (25 mL), and shaken for 5 min. The organic layer was
2
separated, washed with H O (2 ꢆ 50 mL), filtered through anhydrous Na SO , concentrated, and analyzed by HPLC (conditions 2).
2
2
4
HPLC. Conditions 1: ProntoSIL-120-5-C18 AQ column (2 ꢆ 75 mm, # 5 ꢇm; Metrohm AG); mobile phase
CH COONH (100 mM, pH 4.5) (A) and MeCN (B); gradient mode (%B): 0–20 min, 20–26%; flow rate 150 ꢇL/min; column
3
4
temperature 35°C, UV detector at 250 nm. The retention times of standard samples (t , min) were mannose 6.83, glucose
R
1
2.52, and galactose 13.54. Conditions 2: ProntoSIL-120-5-C18 AQ column (2 ꢆ 75 m, # 5 ꢇm; Metrohm AG); mobile phase
LiClO (0.2 M) in HClO (0.006 M) (A) and MeCN (B); gradient mode (%B): 0–9 min, 0–80%; 9–15 min, 80–100%; flow
4
4
rate 100 ꢇL/min; column temperature 35°C; UV detector at 320 and 350 nm. The retention times of standard samples (t , min)
R
were caffeic acid 6.90, p-coumaric acid 8.10, o-coumaric acid 9.73, quercetin 11.48, kaempferol 13.07, isorhamnetin 13.45,
tamarixetin 13.84, and rhamnetin 14.52.
4
4

Chromatographic separation of phenolic compounds from runners of S. betulifolia and S. betulifolia var. aemiliana
used CC over SiO , RP-SiO , polyamide, and Sephadex LH-20 and preparative HPLC as described earlier [9] to isolate from
2
2
S. betulifolia (630 g) 11 compounds that were identified using UV, IR, and NMR spectroscopy and mass spectrometry as 2
(
8 mg), astragalin (5, 12 mg) [22], nicotiflorin (6, 10 mg) [23], tiliroside (7, 63 mg) [24], isoquercitrin (8, 12 mg) [25],
hyperoside (9, 34 mg) [22], avicularin (10, 12 mg) [22], rutin (11, 39 mg) [25], helichrysoside (12, 46 mg) [26], quercetin-3-
O-(6ꢂꢂ-caffeoyl)-ꢀ-D-glucopyranoside (13, 40 mg) [3], and isorhamnetin-3-O-ꢀ-D-glucopyranoside (14, 9 mg) [27].
Chromatographic separation of S. betulifolia var. aemiliana (320 g) isolated 7 compounds that included 2 (6 mg), 7 (21 mg),
8
(5 mg), 9 (11 mg), 11 (18 mg), 12 (14 mg), and 14 (2 mg).
ACKNOWLEDGMENT
The work was sponsored by the RFBR and the Government of the Republic of Buryatia in the framework of Scientific
Project No. 16-43-030857.
REFERENCES
1
.
J. B. Harborne, Comparative Biochemistry of the Flavonoids, Academic Press, London, New York, 1967, 383 pp.
M. I. Choudhary, N. Naheed, A. Abbaskhan, S. Ali, and Atta-ur-Rahman, Phytochemistry, 70, 1467 (2009).
K. Yoshida, A. Hishida, O. Iida, K. Hosokawa, and J. Kawabata, J. Agric. Food Chem., 56, 4367 (2008).
E. A. Karpova and E. P. Khramova, Contemp. Probl. Ecol., 7, 228 (2014).
2
.
3
.
4.
5.
T.-S. Wu, C.-C. Hwang, P.-C. Kuo, T.-H. Kuo, A. G. Damu, and C.-R. Su, Chem. Pharm. Bull., 52, 1227 (2004).
6.
B. T. Ahn, J. G. Kim, J. S. Ro, K. S. Lee, K. J. Oh, S. K. Park, S. G. Chung, and E. H. Cho, Korean J. Pharmacogn.,
27, 178 (1996).
7
8.
.
U. R. Mughal, R. Mehmood, A. Malik, B. Ali, and R. B. Tareen, Helv. Chim. Acta, 95, 100 (2012).
E. A. Karpova and N. P. Lapteva, Turczaninowia, 17, 42 (2014).
9.
D. N. Olennikov and N. I. Kashchenko, Khim. Prir. Soedin., 883 (2017).
1
0.
K. R. Markham, Techniques of Flavonoid Identification, Academic Press, London, New York, 1982, 113 pp.
D. N. Olennikov, L. M. Tankhaeva, and S. V. Agafonova, Appl. Biochem. Microbiol., 47, 419 (2011).
M. Ozipek, I. Calis, M. Ertan, and P. Ruedi, Phytochemistry, 37, 249 (1994).
A. Panighel, M. De Rosso, A. D. Vedova, and R. Flamini, Rapid Commun. Mass Spectrom., 29, 357 (2015).
N. Berardini, R. Fezer, J. Conrad, U. Beifuss, R. Carle, and A. Schieber, J. Agric. Food Chem., 53, 1563 (2005).
D. Gupta and J. Singh, Phytochemistry, 30, 2761 (1991).
1
1.
12.
13.
14.
15.
16.
17.
D. N. Olennikov, N. K. Chirikova, and N. I. Kashchenko, Chem. Nat. Compd., 51, 1085 (2015).
S. S. E. Manga, A. E. Tih, B. Abderamane, R. T. Ghogomu, A. Blond, and B. Bodo, Z. Naturforsch., C: J. Biosci.,
67, 233 (2012).
18.
19.
20.
21.
22.
E. Lee, B.-H. Moon, Y. Park, S. Hong, S. Lee, Y. Lee, and Y. Lim, Bull. Korean Chem. Soc., 29, 507 (2008).
W. Bai, X. Fang, W. Zhao, S. Huang, H. Zhang, and M. Qian, J. Food Drug Anal., 23, 645 (2015).
D. N. Olennikov and N. I. Kashchenko, Chem. Nat. Compd., 50, 633 (2014).
D. N. Olennikov, L. M. Tankhaeva, A. V. Stolbikova, and E. V. Petrov, Chem. Nat. Compd., 47, 165 (2011).
S. S. Azimova and V. I. Vinogradova, Natural Compounds. Flavonoids: Plant Sources, Structure and Properties,
Springer, New York, 2013, p. 199.
23.
24.
25.
26.
27.
D. N. Olennikov, N. I. Kashchenko, and N. K. Chirikova, Molecules, 19, 18296 (2014).
M. Jung and M. Park, Molecules, 12, 2130 (2007).
D. N. Olennikov and V. V. Partilkhaev, J. Planar Chromatogr., 25, 30 (2012).
M. A. Aderogba, L. J. McGaw, M. Bezabih, and B. M. Abegaz, Nat. Prod. Res., 25, 1224 (2011).
D. N. Olennikov and N. I. Kashchenko, Chem. Nat. Compd., 49, 833 (2013).
4
5