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Lovely Goldenrod
(Latin name Solidago
canadensis L.)
|
PhD. Thesis of
Svitlana Dzyubak
Biologically
active substances of several species of Goldenrod genus (Latin
name Solidago)
Table
of Contents
Chapter
1. Species of Goldenrod genus growing in Ukraine; their chemical
composition, chemical and pharmacological properties, usage in folk and
official medicine.
Chapter
2. Preliminary chemical studies and isolation of bioactive
substances.
Chapter
3. Chemical investigation of obtained substances; structure
evaluation.
Chapter
4. Development of polyphenolic medicines with hyponitrogenic activity
(lowering blood urea nitrogen)
from Solidago canadensis.
Chapter
5. Development of combined medicine with hyponitrogenic and diuretic
effects.
|
Abstract
of Dissertation
(Fragments,
part 1)
Chemical
Studies
Scientific
novelty
Three
spices of Goldenrod (Solidago) genus: S. virgaurea, S. canadensis,
and S. alpestris have been studied. Using different methods of
preparative chromatography 48 natural substances were isolated from
these plants. Twenty four were different whereas others were found in
more one species.
Three
previously unknown substances were discovered:
quercetin-3-O-D-(6''-O-acetyl)-glucopyranoside,
kaempferol-3-О-9-D-(6''-О-acetyl)-glucopyranoside and
isorhamnetin-3-О-D-(6''-O-acetyl)-glucopyranoside.
All
chemical investigation of S. alpestris were done for the first time.
Six
substances previously identified in other natural sources, have been
isolated from Solidago
genus for
the first time.
They are ferulic acid (derivative of cinnamic acids), umbelliferone,
scopoletin, scopolin
(derivative of 7-hydroxycoumarin), rhamnetin,
rhamnetin-3-O-D-rutinoside (derivative of flavonoids).
Macro-
and micro-element analyses of polyphenolic and hydrophilic complexes
of S. canadensis, extracted by ethyl alcohol and water respectively,
were obtained for the first time.
Polyphenolic
complex release from mixture with 3 natural adsorbents was studied.
Six sets of experiments measuring the speed of flavonidd release from
a mixture with natural enterosorbents were done for the first time.
Work
description
Investigation
of isolated substances
A
total of forty eight, 24 different, substances were isolated from
three species of Solidago genus: S. virgaurea, S. canadensis and S.
virgaure (table 1). Chemical nature of substances was determined by
using thin-layer chromatography, on silica gel plats. Their
structures have been investigated by spectrophotometric procedures,
Rf-values (table 2) test and other methods (tables 3,4,5,6).
The
following substances, listed in the table under numbers 1-4, were
identified as derivatives of the cinnamic acid: caffeic acid
(3,4-dihydrocinnamic acid), chlorogenic acid (3-caffeoylquinic acid),
isochlorogenic acid (5-caffeoylquinic acid), ferulic acid
(4-hydroxy-3-methoxycinnamic acid).
The
bonding point of caffeic and quinic acids in substance 3 was
determined by obtaining of lactone of quinidin (Fig.1). The
substance's structure was classified as isochlorogenic acid
(5-caffeoylquinic acid). substance 2 did not form lactone of
quinidin, yet, the other characteristics are the same. Based on these
facts, substance 2 was classified as a chlorogenic acid
(3-caffeoylquinic acid).
Fig.1
Chemical transformations of
isochlorogenic acid
Five
substances (5-9), which had blue fluorescence in UV-light on silica
gel plats, were tentatively identified as coumarines
(benzo-alpha-pyrones). The coumarinic nature of substances (5-9) was
proved through their coumarin-producing reaction with hydroiodic acid
dissolved in a mixture of liquefied phenol and acetic anhydride.
Fusion of substance 5 with KOH
yielded 2,4-dihydroxybenzoic acid and resorcin (1,3-dioxybenzene)
(Fig. 2). Methylation of substance 6 by dimethylsulphate yielded 6,
7-dimethoxycoumarin (Fig.
2). Base on data about UV- and IR-spectrum, Rf-values, and the study
of methoxides and acetylates derivatives, substances 5-9 were
identified as: 5 - umbelliferone (7-hydroxycoumarin), 6 - esculetin
(6, 7-dihydroxycoumarin), 8 - scopoletin
(7-hydroxy-6-methoxycoumarin). The hydrolyzation of substances 7 and
9 by 5% HCl in ethyl alcohol-water solution (1:1) yielded
umbelliferone and scopoletin, respectively, as well as glucose.
substances 7 and 9 were concluded to be esculin
(6,7-dihydroxycoumarin 6-glucoside) and scopolin
(scopoletin-7-O-D-glucopyranoside).
Based
on qualitative tests, UV- and IR-spectrum, and Rf-values in various
solvent systems, Briant's cyaniding, and hydrolyzation reactions
substances 10-23 were identified as flavonoids.
Substances
12, 16, and 22 were discovered for the first time.
An
alkaline hydrolysis of substance 12 yielded quercetin-3-O-glucoside
(11) and acetic acid (Fig.3). Both products were verified by
comparative chromatography in ethanol-ammonia (9:1) solution with
standard-substances of quercetin-3-O-glucoside and acetic acid.
Measurement of molecular mass of substances 11 and 12 and
quantitative analysis of the amount of acetyl groups showed that
substance 12 had only one acetyl group. According to the comparative
UV-analysis with diagnostic reagents (0.1 M sodium ethylate and
sodium acetate in ethanol; 4% solutions of boron hydroxide and
aluminum chloride) substance 12, as well as substance 11, were shown
to have hydroxyl groups located on the aglycone in positions 3,5,7,3'
and 4'. This means that the acetate group is attached to glycoside
component.
The
position of the acetic group in substance 12 was establish by
titrating it by sodium metaperiodate. One mole of methanoic acid was
received which allowed to make a conclusion about the presence of an
acetic group in position 6'' of glucose. This fact was also proved by
the measurement of the optical rotation activity of substances 11 and
12 with addition of a copper-ammonia solution.
Fig.2
Chemical transformations of coumarins
Thus,
above mentioned investigations allowed to establish the structure of
substance 12 as
5,7,3',4'-tetrahydroxyflavone-3-O-beta-D-(6''-O-acetyl)-glucopyranoside
or quercetin-3-O-beta-D-(6''-O-acetyl)-glucopyranoside. The structure
of substances 16 and 22 were determined the same way. The chemical
transformation of these substances demonstrated is by the example of
quercetin (Fig. 3).
Among
the products of hydrolysis of substances 13, 17, 19, and 23 were
found D-glucose, L-rhamnose and aglycons: quercetin (10), kaempferol
(14), rhamnetin (18), and isorhamnetin (20). In those substances
sugar components (disaccharides) are located in third position of
flavonoid's aglycone. On the basis of acidic (HCl) and enzymic
hydrolysis (rhamnodiastase), as well as periodate oxidation, the
disaccharides were identified as
glucopyranosyl-(6'')-O-rhamnopyranoside.
Thus
substances 13, 17, 19, and 23 were identified as rutin (quercetin
3-O-rutinoside or 6-O—L-rhamnosyl-D-glucose),
kaempferol-3-О-D-rutinoside, rhamnetin-3-O-D-rutinoside, and
isorhamnetin-3-O-D-rutinoside respectively.
Identification
of substance 24 was made on basis of physicochemical characteristics
of the substance itself and its derivatives, as well as its
IR-spectrum. They were also tested by the melting point depression
test. Based on this data substance 24 was identified as oleanolic
acid (3-beta-hydroxyolean-12-en-28-oic acid).
In
addition to qualitative analysis, a quantitative one was done. The
total amount of flavanoids in S. alpestris is 3.5%, in S. canadensis
– 5.6%, in S. virgaurea -4.5%.
Based
on semiquantitative comparative studies mentioned above, it has been
shown that quercetin and kaempferol groups are more common in S.
canadensis and S. virgaurea whereas S. alpestris
contained more flavanoids of isorhamnetin group.
Results
of these studies allowed to recommend S. canadensis and S. virgaurea
as raw material for the development of new herbal medicine for
treatment of kidney diseases. Data about chemical content of S.
alpestris was found for the first time.
Fig.3 Chemical transformations of flavonoids
Substances
extracted from herbs of Goldenrod genus (Lati name
Solidago L.)
Table 1.
#
|
Name
|
Formula
|
Melting
point
(OС)
|
Optical rotation |
Sources
|
|
|
Derivatives
of cinnamic acids
|
|
|
|
1
|
Caffeic
acid
(3,4-dihydrocinnamic acid) |
|
194-195
|
|
S.virgaurea
L.,
S.alpestris
Waldst. et Kit., S.canadensis
L. |
2
|
Chlorogenic
acid
(3-Caffeoylquinic acid) |
|
201-203
|
-32.3
|
S.virgaurea
L.,
S.alpestris
Waldst. et Kit.,
S.canadensis
L. |
3
|
Isochlorogenic
acid (5-Caffeoylquinic acid) |
|
200-201
|
|
S.virgaurea
L.,
S.canadensis
L. |
4
|
Ferulic acid
(4-hydroxy-3-methoxycinnamic acid)
|
|
167-169
|
|
S.canadensis
L. |
|
|
Coumarins
(Benzo-a-pyrones) and their derivatives
|
|
|
|
5
|
Umbelliferone
(7-hydroxycoumarin)
|
|
234-235
|
|
S.canadensis
L. |
6
|
Esculetin
(6,
7-dihydroxycoumarin) |
|
269-272
|
|
S.virgaurea
L.,
S.canadensis
L. |
7
|
Esculin
(6,7-dihydroxycoumarin 6-glucoside) |
|
203-206
|
-146
(methanol)
|
S.virgaurea
L.,
S.canadensis
L. |
8
|
Scopoletin
(7-Hydroxy-6-methoxycoumarin) |
|
204-205
|
|
S.alpestris
Waldst. et Kit.,
S.canadensis
L. |
9
|
Scopolin
(scopoletin-7-O-D-glucopyranoside) |
|
218-220
|
-8.5
(dimethylformamide)
|
S.canadensis
L. |
|
|
Flavonoids
(2-Phenylbenzo--pyrons) and their derivatives
|
|
|
|
10
|
Quercetin
(3,3',4'5,7-pentahydroxy flavone) |
|
311-314
|
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
11
|
Isoquercitrin
(quercetin-3-O-glucoside)
|
|
218-220
|
-
36
(methanol)
|
S.virgaurea L.,
S.canadensis
L. |
12
|
Quercetin-3-O-D-(6''-O-acetyl)-glucopyranoside
|
|
201-203
|
|
S.canadensis
L. |
13
|
Rutin
(quercetin
3-O-rutinoside or 6-O--L-rhamnosyl-D-glucose) |
|
188-192
|
-
32
(dimethylformamide)
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
14
|
Kaempferol
(3,4',5,7-tetrahydroxyflavone) |
|
283-285
|
|
S.virgaurea
L.,
S.canadensis
L. |
15
|
Kaempferol-3-O-D-glucopyranoside
|
|
210-213
|
-
16
(methanol)
|
S.virgaurea
L.,
S.canadensis
L. |
16
|
Kaempferol-3-О-9-D-(6''-О-acetyl)-glucopyranoside
|
|
200-201
|
|
S.canadensis
L. |
17
|
Kaempferol-3-О-D-rutinoside
|
|
188-192
|
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
18
|
Rhamnetin
(3,5,3',4'-Tetrahydroxy-7-methoxyflavone) |
|
296-298
|
|
S.virgaurea
L.,
S.canadensis
L. |
19
|
Rhamnetin-3-O-D-rutinoside
|
|
184-187
|
-
43.4
(dimethylformamide)
|
S.virgaurea
L.,
S.canadensis
L. |
20
|
Isorhamnetin
(3,5,7,4'-tetrahydroxy-3'-methoxyflavone) |
|
314-316
|
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
21
|
Isorhamnetin-3-O-D-glucopyranoside
|
|
250-251
|
- 59
(dimethylformamide)
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
22
|
Isorhamnetin-3-О-D-(6''-O-acetyl)-glucopyranoside
|
|
198-202
|
- 28
(dimethylformamide)
|
S.canadensis
L. |
23
|
Isorhamnetin-3-O-D-rutinoside
|
|
171-173
|
-
32.2
|
S.virgaurea
L.,
S.alpestris
Waldst.et Kit.,
S.canadensis
L. |
|
|
Sapogenin
(triterpene)
|
|
|
|
24
|
Oleanolic acid
(3-beta-hydroxyolean-12-en-28-oic acid) |
|
306-308
|
+80.0
(methanol)
|
S.canadensis
L. |
Several
characteristics of isolated substances Table
2 (*.pdf, 282 KB).
Rf-value
in different solvents (paper and silica gel plates) Table 2+3 (*.pdf, 553
KB).
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