A tetra-acylated cyanidin 3-sophoroside-5-glucoside from the purple-violet flowers of Moricandia arvensis (L.) DC. (Brassicaceae)

Article abstract of DOI:10.1016/j.phytol.2012.12.007

A novel tetra-acylated cyanidin 3-sophoroside-5-glucoside was isolated from the purple-violet flowers of Moricandia arvensis (L.) DC. (Family: Brassicaceae), and determined to be cyanidin 3-O-[2-O-(2-O-(4-O-(6-O-(4-O- (β-glucopyranosyl)-trans-caffeoyl)-β-glucopyranosyl)-trans-caffeoyl) -β-glucopyranosyl)-6-O-(trans-caffeoyl)-β-glucopyranoside] -5-O-[6-O-(malonyl)-β-glucopyranoside] by chemical and spectroscopic methods.

Full text of DOI:10.1016/j.phytol.2012.12.007

Phytochemistry Letters 6 (2013) 170–173
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Phytochemistry Letters
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A tetra-acylated cyanidin 3-sophoroside-5-glucoside from the purple-violet
flowers of Moricandia arvensis (L.) DC. (Brassicaceae)
a
a
b
a
a
Fumi Tatsuzawa a, , Shun Ito , Motoki Sato , Hiroki Muraoka , Kazuhisa Kato , Yoshihito Takahata ,
*
Satoshi Ogawa b
a Faculty of Agriculture, Iwate University, Morioka, Iwate 020-8550, Japan
b Faculty of Engineering, Iwate University, Morioka, Iwate 020-8551, Japan
A R T I C L E I N F O
A B S T R A C T
Article history:
A novel tetra-acylated cyanidin 3-sophoroside-5-glucoside was isolated from the purple-violet flowers
Received 5 November 2012
Received in revised form 11 December 2012
Accepted 21 December 2012
Available online 14 January 2013
of Moricandia arvensis (L.) DC. (Family: Brassicaceae), and determined to be cyanidin 3-O-[2-O-(2-O-(4-
O-(6-O-(4-O-(
6-O-(trans-caffeoyl)-
spectroscopic methods.
b
-glucopyranosyl)-trans-caffeoyl)-
b
-glucopyranosyl)-trans-caffeoyl)-
b-glucopyranosyl)-
b
-glucopyranoside]-5-O-[6-O-(malonyl)-
b
-glucopyranoside] by chemical and
ß 2013 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.
Keywords:
Moricandia arvensis (L.) DC.
Brassicaceae
Purple-violet flower color
Tetra-acylated cyanidin 3-sophoroside-5-
glucoside
1. Introduction
restricted to the flowers of the genera Raphanus, Iberis and
Moricandia (Saito et al., 2008; Tatsuzawa et al., 2008b, 2012a). In
The genus Moricandia DC. (Brassicaceae), includes eight species
of annual or perennial herbs that are native to the Mediterranean
region and also cultivated as ornamental plants with purple, violet,
and white flowers, in Europe and America. The leaves and/or stems
of Moricandia arvensis (L.) DC. (Violet Cabbage) are used in
traditional cookery and the treatment of syphilis in Tunisia
(Braham et al., 2005). In the limited flavonoid work reported for
this species, flavonol glycosides have been characterized (Braham
et al., 2005). Moreover, floral anthocyanins in the genus Moricandia
have only been characterized from the purple flowers of
Moricandia ramburii (Tatsuzawa et al., 2012a), and not from the
purple-violet flowers of M. arvensis.
this paper, the structure elucidation of a new tetra-acylated
cyanidin 3-sophoroside-5-glucoside from the purple-violet flow-
ers of M. arvensis is reported.
2. Results and discussion
More than 10 anthocyanins were detected in a methanol–acetic
acid–water (MAW) (4:1:5, v/v/v) extract from purple-violet
flowers of M. arvensis by high performance liquid chromatography
(HPLC) analysis. The percentage of the major pigment (1),
calculated as the total anthocyanin content by HPLC peak area
at 530 nm, was 25.3%.
The major pigment (1) was extracted from the purple-violet
flowers with 5% HOAc, followed by isolation using Diaion HP-20
(Mitsubishi Chemical’s Ion Exchange Resins, aromatic type
adsorbent) column chromatography (CC), preparative HPLC and
thin layer chromatography (TLC) (Tatsuzawa et al., 2012a).
Acid hydrolysis of 1 yielded cyanidin as its anthocyanidin
(Harborne, 1984). Moreover, trans-caffeic acid and malonic acid
were detected in the hydrolysates of 1 by HPLC.
To date, 20 acylated 3-sophoroside-5-glucosides of pelargoni-
din, cyanidin and peonidin isolated from 3 genera and 50 acylated
3-sambubioside-5-glucosides of pelargonidin, cyanidin and del-
phinidin isolated from
9 genera have been unambiguously
determined from the flowers of Brassicaceae (Honda et al.,
2005; Saito et al., 1995a, 1996, 2008, 2011; Tatsuzawa, 2012;
Tatsuzawa et al., 2006, 2007, 2008a, b, 2010, 2012a, b, c). The
distribution of 3-sophoroside-5-glucosides of anthocyanidins is
Alkaline hydrolysis of 1 yielded cyanidin 3-sophoroside-5-
glucoside, 4-O-caffeoyl-glucoside and malonic acid. The deacyl
anthocyanin and 4-O-caffeoyl-glucoside structures were identified
via co-HPLC with authentic cyanidin 3-sophoroside-5-glucoside
* Corresponding author. Tel.: +81 19 621 6145; fax: +81 19 621 6145.
E-mail address: fumi@iwate-u.ac.jp (F. Tatsuzawa).
1874-3900/$ – see front matter ß 2013 Phytochemical Society of Europe. Published by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.phytol.2012.12.007

F. Tatsuzawa et al. / Phytochemistry Letters 6 (2013) 170–173
171
and 4-O-caffeoyl-glucoside, prepared from Ipomoea purpurea
(Saito et al., 1995b) by alkaline hydrolysis.
coupling constants were assigned as shown in Table 1. Three sets
of two pairs of doublet resonances, assigned to the six olefinic
proton signals of the caffeic acid moieties, indicated trans
configuration for these acids based on their coupling constants
(J = 15.8 Hz each) (Table 1). The proton chemical shifts of the
2.1. Pigment 1
The molecular ion [M]+ of 1 was observed at m/z 1669
(C75H81O43),indicatingonemoleculeofcyanidinwithfivemolecules
of glucose, three molecules of caffeic acid and one molecule of
malonic acid. The elemental components of pigment 1 were
confirmed by measuring its high resolution FABMS (Section 4.4.1).
sugar moieties were observed in the region of
dH 5.61–3.02, with
the five anomeric proton resonances at H 5.61 (d, J = 7.3 Hz, Glc
d
A), 5.14 (d, J = 7.6 Hz, Glc B), 5.15 (d, J = 7.6 Hz, Glc C), 4.85 (d,
J = 7.3 Hz, Glc D) and 4.79 (d, J = 8.3 Hz, Glc E). Based on the
observed coupling constants (Table 1), these five sugars were
The structure of pigment
1
was further elucidated by
assumed to be in b-pyranose form. The linkages and/or positions
of the attachments of the sugar and acyl groups were determined
based on 2D COSY and 2D NOESY experiments.
investigation of its 1H (500 MHz) and 13C (126 MHz) NMR spectra,
including 2D COSY, 2D NOESY (with 300 ms mixing time), 1H–13
C
HMQC and 1H–13C HMBC spectra in DMSO-d6–CF3COOD (9:1)
(Table 1 and Fig. 1).
By the application of its NOESY experiment, NOEs between
H-1 of Glc A and H-4 (
H 6.94) of cyanidin, and H-2 (
were observed (Fig. 1), supporting the glycosylation of the C-3
d
H 8.72) of cyanidin, H-1 of Glc B and H-6
In the 1H NMR spectrum, the chemical shifts of 15 aromatic
protons of the cyanidin and caffeic acid moieties with their
(
d
d
H 4.12) of Glc A and H-1 of Glc C
Table 1
NMR spectroscopic data of pigment 1 from the flowers of Moricandia arvensis in DMSO-d6/CF3CO2D (9:1).
1H (ppm)
13C (ppm)
1H (ppm)
13C (ppm)
Cyanidin
Glucose E
2
162.9
145.9
132.2
155.4
105.0
167.8
96.5
1
4.79 d(8.3)
3.37 t(7.9)
3.22 t(9.1)
3.20 t(8.9)
3.40 m
102.3
73.7
74.1
70.3
74.7
61.2
3
2
4
8.72 s
6.95 s
6.94 s
3
5
4
6
5
7
6a
6b
3.69 m
8
3.72 dd(1.3, 11.8)
9
155.3
111.9
120.1
117.4
146.9
155.7
116.9
128.8
10
10
20
30
40
50
60
Caffeic acid (I)
1
126.0
116.0
145.7
148.6
116.2
121.3
114.1
145.7
167.1
7.93 d(2.3)
2
6.83 d(1.6)
3
4
7.12 d(8.8)
5
6.67 d(8.2)
8.33 dd(2.3, 8.8)
6
a
b
COOH
6.77 dd(1.6, 8.2)
6.13 d(15.8)
7.26 d(15.8)
Glucose A
1
5.61 d(7.3)
4.12 t(8.0)
99.1
77.7
76.9
70.3
73.7
63.3
2
3
3.60 t(8.2)
Caffeic acid (II)
4
3.42 t(8.2)
1
129.6
115.4
147.4
147.7
116.1
121.3
117.4
144.9
166.2
5
3.91 m
2
7.15 d(1.3)
6a
6b
4.23 dd(5.8, 11.7)
4.36 brd(11.7)
3
4
5
7.08 d(8.5)
Glucose B
6
6.97 dd(1.3, 8.5)
6.36 d(15.8)
7.43 d(15.8)
1
5.14 d(7.6)
3.52 t(8.9)
102.0
73.6
76.2
70.3
74.7
64.4
a
b
COOH
2
3
3.37 t(7.9)
4
3.24 t(8.8)
5
3.74 m
Caffeic acid (III)
6a
6b
4.06 dd(6.1, 11.5)
4.41 brd(11.5)
1
129.4
115.6
147.5
148.0
115.4
121.5
116.5
145.3
166.9
2
7.18 d(1.6)
3
Glucose C
4
1
5.15 d(7.6)
4.63 t(8.8)
3.37 t(7.9)
3.16 t(9.3)
3.02 m
100.4
75.0
76.2
70.3
77.8
61.6
5
7.14 d(8.5)
2
6
a
b
COOH
7.10 dd(1.6, 8.5)
6.43 d(15.8)
7.54 d(15.8)
3
4
5
6a
6b
3.41 m
3.60 brd(11.1)
Malonic acid
CH2
3.32 s
41.6
167.3
168.5
Glucose D
COOH
1
4.85 d(7.3)
3.40 m
102.1
73.7
75.9
70.3
74.7
63.8
COOH
2
3
3.32 m
4
3.31 m
5
3.71 m
6a
6b
4.27 dd(5.7, 11.7)
4.45 brd(11.7)

172
F. Tatsuzawa et al. / Phytochemistry Letters 6 (2013) 170–173
of xylose moiety in their 3-sambubioside residue (Honda et al.,
2005). These complicated structures of acyl groups are considered
to be an advanced character in the Brassicaceae (Harborne, 1967;
Honda and Saito, 2002).
4. Experimental
4.1. General procedures
TLC was carried out on plastic coated cellulose sheets (Merck)
using seven mobile phases: BAW (n-BuOH–HOAc–H2O, 4:1:2, v/v/
v), BuHCl (n-BuOH–2 N HCl, 1:1, v/v, upper layer), AHW (HOAc–
HCl–H2O, 15:3:82, v/v/v), and 1% HCl for anthocyanins, Forestal
(HOAc–HCl–H2O, 30:3:10, v/v/v) for anthocyanidins and BAW, EAA
(EtOAc–HOAc–H2O, 3:1:1, v/v/v) and EFW (EtOAc–HCOOH–H2O,
5:2:1, v/v/v) for sugars and organic acids with UV light and aniline
hydrogen phthalate spray reagent (Harborne, 1984).
Fig. 1. Structure of pigment 1 isolated from the purple-violet flowers of Moricandia
arvensis. Observed NOEs are indicated by arrows.
and C-5 cyanidin hydroxyl groups with Glc A and Glc B,
respectively, while also indicating
between Glc A and Glc C.
a
sophorose structure
Analytical HPLC was performed on a LC 10A system (Shimadzu),
using a Waters C18 (4.6 mm ꢀ 250 mm) column at 40 8C with a
flow rate of 1 mL/min and monitoring at 530 nm. The eluant was
applied as a linear gradient elution for 40 min from 20 to 85%
solvent B (1.5% H3PO4, 20% HOAc, 25% MeCN in H2O) in solvent A
(1.5% H3PO4 in H2O) with 5 min of re-equilibration at 20% solvent
B, for anthocyanins, anthocyanidins and hydroxycinnamic acids
(method 1). The other eluant for malonic acid was applied as an
isocratic elution of solvent A for 10 min and monitored at 210 nm
(Tatsuzawa et al., 2009) (method 2).
UV–vis spectra were recorded on UV–vis Multi Purpose
Spectrophotometer (MPS-2450, Shimadzu) in 0.1% HCl–MeOH
(from 200 to 700 nm).
High resolution FAB mass (FABMS) spectra were determined on
a JEOL JMS-700 Mass spectrometer operating in the positive ion
mode using 1:1 mixture of dithiothreitol and 3-nitrobenzyl alcohol
as a matrix. 1H (500 MHz) and 13C (126 MHz) NMR spectra were
measured on a Bruker Avance III 500 MHz NMR spectrometer using
CF3CO2D-DMSO-d6 (1:9) as a solvent. Chemical shifts are reported
Six characteristic downfield shifted proton signals were
assigned to the methylene protons of Glc A ( H 4.23 and 4.36, H-
6a and b), Glc B ( H 4.06 and 4.41, H-6a and b), Glc D ( H 4.27 and
4.45, H-6a and b) and to a methine proton ( H 4.63, t, J = 8.8 Hz, H-
d
d
d
d
2) of Glc C, indicating acylations of C-6 OH of Glc A, Glc B and Glc D
and C-2 OH of Glc C with four acid molecules. In the NOESY
spectrum, the correlations between H-6a,b of Glc A and H-
caffeic acid (I), H-2 of Glc C and H-
Glc D and H-5,6 of caffeic acid (II), H-6a,b of Glc D and H-
a
,
b
of
a
,
b
of caffeic acid (II), H-1 of
a
,
b
of
caffeic acid (III) and H-1 of Glc E and H-5,6 of caffeic acid (III)
were observed, establishing the acylation groups at C-6 OH of
(Glc A and D), C-2 OH (Glc C) and C-1 OH (Glc D and E) as caffeic
acid (I and III), caffeic acid (II) and caffeic acid (II and III),
respectively. Moreover, the methylene proton signals of Glc B
were observed at 4.01 and 4.10 ppm in a lower magnetic field.
Thus, OH-6 of Glc B was assumed to be acylated with malonic
acid.
Consequently, the structure of pigment 1 was determined to be
relative to DMSO-d6 (dH 2.50 ppm, dC 39.5 ppm), and coupling
cyanidin 3-O-[2-O-(2-O-(4-O-(6-O-(4-O-(
caffeoyl)- -glucopyranosyl)-trans-caffeoyl)-
O-(trans-caffeoyl)- -glucopyranoside]-5-O-[6-O-(malonyl)-
b
-glucopyranosyl)-trans-
-glucopyranosyl)-6-
-glu-
constants (J) are in Hz.
b
b
b
b
4.2. Plant materials
copyranoside], which is a new anthocyanin in plants (Andersen and
Jordheim, 2006; Harborne and Baxter, 1999; Honda and Saito, 2002;
Veitch and Grayer, 2008, 2011). The 13C spectrum of pigment 1 was
completely assigned using the HMQC and HMBC experiments
(Table 1).
The seeds of M. arvensis (L.) DC. (accession number: MOR-ARV-
8) were obtained from Prof. T. Nishio at the Tohoku University
Brassica Seed Bank, and grown in greenhouses at the Laboratory of
Olericultural and Floricultural Science, Faculty of Agriculture,
Iwate University. The flowers exhibited purple-violet [purple-
violet 82C by Royal Horticultural Society color chart and
chromaticity value, b*(ꢁ30.41)/a*(58.56)]. Flowers were collected
from winter to spring seasons in Iwate, Japan and dried overnight
at 40 8C, and kept in a refrigerator at 4 8C. The chromaticity values
were recorded on a NR-3000 handy colorimeter (Nippon Denshoku
Industries Co., Ltd.).
3. Concluding remarks
A new acylated cyanidin glycoside was isolated from the
purple-violet flowers of M. arvensis. From the chemotaxonomical
point of view, there are two C-3 OH glycosidic patterns for
anthocyanidins found in the flowers of Brassicaceae species,
namely acylated 3-sophoroside from Iberis umbellata, M. ramburii
and Raphanus sativus (Saito et al., 2008; Tatsuzawa et al., 2008b,
2012a) and acylated 3-sambubioside from Aubrieta ꢀ cultorum,
Cheiranthus cheiri, Heliophila coronopifolia, Hesperis matronalis,
Lobularia maritima, Lunaria annua, Malcolmia maritima, Matthiola
incana and Orychophragmus violaceus (Honda et al., 2005; Saito
et al., 1995a, 1996, 2011; Tatsuzawa, 2012; Tatsuzawa et al., 2006,
2007, 2008a, 2010, 2012a, b, c). Therefore, the floral anthocyanins
of M. arvensis are grouped into the former pattern.
4.3. Isolation of anthocyanin
Dried flowers (100 g) of M. arvensis were immersed in HOAc–
H2O (3 L; HOAc–H2O, 1:19) at room temp. for 5 h and extracted.
The extract was passed through a Diaion HP-20 (Mitsubishi
Chemical’s Ion Exchange Resins, aromatic type adsorbent) column
(90 mm ꢀ 150 mm), on which acylated anthocyanins were
adsorbed. The column was thoroughly washed with H2O (2 L)
and eluted with 5% HOAc–MeOH (500 mL) to recover the
anthocyanins. After concentration, the residue was separated
and purified with paper chromatography (PC) using BAW. The
separated pigment was further purified by TLC (15% HOAc) and
The acylated pattern of pigment 1 is acylated at the 2-position
of outer glucose in the 3-sophoroside with a long-linear side chain
of glucosyl caffeoyl-glucosyl-caffeic acid. This long-linear side
chain is similar to those of Orychophragmus violet-blue antho-
cyanins whose long-linear side chains were acylated at the C-2 OH
prep. HPLC. Prep. HPLC was performed on
a Waters C18

F. Tatsuzawa et al. / Phytochemistry Letters 6 (2013) 170–173
173
(19 mm ꢀ 150 mm) column at 40 8C with a flow rate of 4 mL/min
Acknowledgements
and monitoring at 530 nm. The solvent used was as follows: a
linear gradient elution for 30 min from 50 to 60% solvent B in
solvent A. The fraction was transformed to a Diaion HP-20 column,
on which pigment was adsorbed. Anthocyanin pigment was eluted
with 5% HOAc–MeOH followed by addition of excess of Et2O, and
then dried. The purified pigment from the flowers was obtained as
pigment 1 (ca. 23 mg).
We thank Prof. T. Nishio (Tohoku University) for the donation of
M. arvensis seeds (accession number: MOR-ARV-8: Tohoku
University Brassica Seed Bank) and Mr. James Hall (Iwate
University) and Dr. Norio Saito (Meiji-Gakuin University) for
careful revision of the manuscript.
This work was supported in part by a Grant-in-Aid for Science
Research (C) [No. 22580024, to F.T.] from the Japan Society for the
Promotion of Science (JSPS).
4.4. Analyses of anthocyanins
References
The identification of pigment 1 was carried out by standard
procedures with both alkaline and acid hydrolyses (Harborne,
1984). Acid hydrolysis of 1 (ca. 1 mg) was carried out with 2 N HCl
(1 mL) at 100 8C for 1 h. Alkaline hydrolysis of 1 (ca. 1 mg) was
carried out with 2 N NaOH solution (1 mL) under degassed syringe
allowed to stand for 15 min. The solution was next acidified with
2 N HCl (1.1 mL) and evaporated in vacuo to dryness. These
products were analyzed by standard procedures (Harborne, 1984).
Andersen, Ø.M., Jordheim, M., 2006. The anthocyanins. In: Andersen, Ø.M., Mark-
ham, K.R. (Eds.), Flavonoids: Chemistry, Biochemistry and Applications. CRC
Press, Boca Raton, pp. 471–551.
Braham, H., Mighri, Z., Jannet, H.B., Matthew, S., Abreu, P.M., 2005. Antioxidant
phenolic glycosides from Moricandia arvensis. J. Nat. Prod. 68, 517–522.
Harborne, J.B., 1967. Comparative Biochemistry of Flavonoids. Academic Press,
London/New York.
Harborne, J.B., 1984. Phytochemical Methods, 2nd ed. Chapman and Hall, London.
Harborne, J.B., Baxter, H., 1999. The Handbook of Natural Flavonoids, vol. 2. John
Wiley & Sons, Chichester.
The data of TLC (Rf values), HPLC (Rt-min, method 1), UV–vis (lmax),
Honda, T., Saito, N., 2002. Recent progress in the chemistry of polyacylated antho-
cyanins as flower color pigments. Heterocycles 56, 633–692.
and FABMS spectra are shown in Section 4.4.1.
Honda, T., Tatsuzawa, F., Kobayashi, N., Kasai, H., Nagumo, S., Shigihara, A., Saito, N.,
2005. Acylated anthocyanins from the violet-blue flowers of Orychophragonus
violaceus. Phytochemistry 66, 1844–1851.
Saito, N., Tatsuzawa, F., Nishiyama, A., Yokoi, M., Shigihara, A., Honda, T., 1995a.
Acylated cyanidin 3-sambubioside-5-glucoside in Matthiola incana. Phyto-
chemistry 38, 1027–1032.
Saito, N., Tatsuzawa, F., Yoda, K., Yokoi, M., Kasahara, K., Iida, S., Shigihara, A., Honda,
T., 1995b. Acylated cyanidin glycosides in the violet-blue flowers of Ipomoea
purpurea. Phytochemistry 40, 1283–1289.
4.4.1. Pigment 1
Dark purple-red powders; UV–vis (in 0.1% HCl–MeOH): lmax
533, 321, 286 nm, Eacyl/Emax = 132%, E440/Emax = 17%, AlCl3 shift +;
TLC: Rf-values (100ꢀ) BAW 20, BuHCl 13, 1% HCl 32, AHW 72;
HPLC: Rt (min) 28.3; HR-FABMS calc. for C75H81O43: 1669.4152.
Found: 1669.4095.
Saito, N., Tatsuzawa, F., Hongo, A., Win, K.W., Yokoi, M., Shigihara, A., Honda, T.,
1996. Acylated pelargonidin 3-sambubioside-5-glucoside in Matthiola incana.
Phytochemistry 41, 1613–1630.
Saito, N., Tatsuzawa, F., Suenaga, E., Toki, K., Shinoda, K., Shigihara, A., Honda, T.,
2008. Tetra-acylated cyanidin 3-sophoroside-5-glucosides from the flowers of
Iberis umbellata L. (Cruciferae). Phytochemistry 69, 3139–3150.
Saito, N., Tatsuzawa, F., Toki, K., Shinoda, K., Shigihara, A., Honda, T., 2011. The blue
anthocyanin pigments from the blue flowers of Heliophila coronopifolia L.
(Brassicaceae). Phytochemistry 72, 2219–2229.
4.5. Analyses of hydrolysates (deacylanthocyanins, anthocyanidins,
sugar and acids)
The identification of hydrolysates of pigment 1 by alkaline and
acid, was carried out by standard procedures (Harborne, 1984). The
results were as follows.
Tatsuzawa, F., 2012. Acylated cyanidin 3-sambubioside-5-glucosides in the purple
flowers of Hesperis matronalis L. (Brassicaceae). Biochem. Syst. Ecol. 44, 374–379.
Tatsuzawa, F., Saito, N., Shinoda, K., Shigihara, A., Honda, T., 2006. Acylated cyanidin
3-sambubioside-5-glucosides in three garden plants of the Cruciferae. Phyto-
chemistry 67, 1287–1295.
Tatsuzawa, F., Toki, K., Saito, N., Shinoda, K., Shigihara, A., Honda, T., 2007. Four
acylated cyanidin 3-sambubioside-5-glucosides from the purple-violet flowers
of Lobularia maritima. Heterocycles 71, 1117–1135.
4.5.1. Cyanidin 3-sophoroside-5-glucoside
UV–vis: lmax 525, 279 nm, E440/Emax = 13%, AlCl3 shift +; TLC:
Rf-values (100ꢀ) BAW 20, BuHCl 10, 1% HCl 45, AHW 75; HPLC
(method 1): Rt (min) 12.5.
Tatsuzawa, F., Saito, N., Toki, K., Shinoda, K., Shigihara, A., Honda, T., 2008a.
Triacylated cyanidin 3-(3X-glucosylsambubioside)-5-glucosides from the flow-
ers of Malcolmia maritima. Phytochemistry 69, 1033–1040.
Tatsuzawa, F., Toki, K., Saito, N., Shinoda, K., Shigihara, A., Honda, T., 2008b.
Anthocyanin occurrence in the root peels petioles and flowers of red radish
(Raphanus sativus L.). Dyes Pigments 79, 83–88.
Tatsuzawa, F., Saito, N., Mikanagi, Y., Shinoda, K., Toki, K., Shigihara, A., Honda, T.,
2009. An unusual acylated malvidin 3-glucoside from flowers of Impatiens
textori Miq. (Balsaminaceae). Phytochemistry 70, 672–674.
4.5.2. Cyanidin
UV–vis: lmax 536, 273 nm, E440/Emax = 44%, AlCl3 shift +; TLC:
Rf-values (100ꢀ) Forestal 42; HPLC (method 1): Rt (min) 25.3.
4.5.3. Glucose
TLC: Rf-values (100ꢀ) BAW 24, EAA 18, EFW 49; Color (AHP)
Brown.
Tatsuzawa, F., Usuki, R., Toki, K., Saito, N., Shinoda, K., Shigihara, A., Honda, T., 2010.
Acylated pelargonidin 3-sambubioside-5-glucosides from the red-purple flow-
ers of Lobularia maritima. J. Jpn. Soc. Hort. Sci. 79, 84–90.
4.5.4. Caffeic acid
UV: lmax 325, 297 (in MeOH), TLC: Rf-values (100ꢀ) BAW 82,
EAA 92, EFW 73 Color (under UV) Blue; HPLC (method 1): Rt (min)
12.0.
Tatsuzawa, F., Ito, K., Muraoka, H., Namauo, T., Kato, K., Takahata, Y., Ogawa, S.,
2012a. Triacylated peonidin 3-sophoroside-5-glucosides from the purple flow-
ers of Moricandia ramburii Webb. Phytochemistry 76, 73–77.
Tatsuzawa, F., Saito, N., Toki, K., Shinoda, K., Honda, T., 2012b. Flower colors and their
anthocyanins in Matthiola incana cultivars (Brassicaceae). J. Jpn. Soc. Hort. Sci. 81,
91–100.
4.5.5. 4-O-glucosyl-caffeic acid
Tatsuzawa, F., Aiba, Y., Morino, T., Saito, N., Shinoda, K., Kato, K., Toki, K., Honda, T.,
2012c. Copigmentation with acylated anthocyanin and kaempferol glycosides
in violet and purple flower cultivars of Aubrieta ꢀ cultorum (Brassicaceae). J. Jpn.
Soc. Hort. Sci. 81, 275–284.
Veitch, N.C., Grayer, R.J., 2008. Flavonoids and their glycosides, including antho-
cyanins. Nat. Prod. Rep. 25, 555–611.
UV: lmax 316, 288 (in MeOH), TLC: Rf-values (100ꢀ) BAW 63,
EAA 81, EFW 55; Color (under UV) Blue; HPLC (method 1): Rt (min)
8.5.
4.5.6. Malonic acid
Veitch, N.C., Grayer, R.J., 2011. Flavonoids and their glycosides, including antho-
cyanins. Nat. Prod. Rep. 28, 1626–1695.
HPLC (method 2): Rt (min) 4.1.