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Polysaccharides of Salsola passerina: Extraction, Structural Characterization and Antioxidant Activity


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The above-ground part of the Salsola passerine was found to contain ~13% (w/w) of polysaccharides extractable with water and aqueous solutions of ammonium oxalate and sodium carbonate. The fractions extracted with aqueous sodium carbonate solutions had the highest yield. The polysaccharides of majority fractions are characterized by similar monosaccharide composition; namely, galacturonic acid and arabinose residues are the principal components of their carbohydrate chains. The present study focused on the determination of antioxidant activity of the extracted polysaccharide fractions and elucidation of the structure of polysaccharides using nuclear magnetic resonance (NMR) spectroscopy. Homogalacturonan (HG), consisting of 1,4-linked residues of α-D-galactopyranosyluronic acid (GalpA), rhamnogalacturonan-I (RG-I), which contains a diglycosyl repeating unit with a strictly alternating sequence of 1,4-linked D-GalpA and 1,2-linked L-rhamnopyranose (Rhap) residues in the backbone, and arabinan, were identified as the structural units of the obtained polysaccharides. HMBC spectra showed that arabinan consisted of alternating regions formed by 3,5-substituted and 1,5-linked arabinofuranose residues, but there was no alternation of these residues in the arabinan structure. Polysaccharide fractions scavenged the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical at 0.2–1.8 mg/mL. The correlation analysis showed that the DPPH scavenging activity of polysaccharide fractions was associated with the content of phenolic compounds (PCs).
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Int. J. Mol. Sci. 2022, 23, 13175.
Polysaccharides of Salsola passerina: Extraction, Structural
Characterization and Antioxidant Activity
Victoria Golovchenko
, Sergey Popov
*, Vasily Smirnov
, Victor Khlopin
, Fedor Vityazev
Shinen Naranmandakh
, Andrey S. Dmitrenok
and Alexander S. Shashkov
Institute of Physiology of Federal Research Centre “Komi Science Centre of the Urals Branch of the Russian
Academy of Sciences”, 50 Pervomaiskaya Str., 167982 Syktyvkar, Russia
School of Arts and Sciences, National University of Mongolia, Baga Toiruu 47, Ulaanbaatar 14201, Mongolia
N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, 47, Leninsky Prospect,
119991 Moscow, Russia
* Correspondence:; Tel./Fax: +78-21-224-1001
Abstract: The above-ground part of the Salsola passerine was found to contain ~13% (w/w) of poly-
saccharides extractable with water and aqueous solutions of ammonium oxalate and sodium car-
bonate. The fractions extracted with aqueous sodium carbonate solutions had the highest yield. The
polysaccharides of majority fractions are characterized by similar monosaccharide composition;
namely, galacturonic acid and arabinose residues are the principal components of their carbohy-
drate chains. The present study focused on the determination of antioxidant activity of the extracted
polysaccharide fractions and elucidation of the structure of polysaccharides using nuclear magnetic
resonance (NMR) spectroscopy. Homogalacturonan (HG), consisting of 1,4-linked residues of α-D-
galactopyranosyluronic acid (GalpA), rhamnogalacturonan-I (RG-I), which contains a diglycosyl
repeating unit with a strictly alternating sequence of 1,4-linked D-GalpA and 1,2-linked L-rhamno-
pyranose (Rhap) residues in the backbone, and arabinan, were identified as the structural units of
the obtained polysaccharides. HMBC spectra showed that arabinan consisted of alternating regions
formed by 3,5-substituted and 1,5-linked arabinofuranose residues, but there was no alternation of
these residues in the arabinan structure. Polysaccharide fractions scavenged the 1,1-diphenyl-2-pic-
rylhydrazyl (DPPH) radical at 0.2–1.8 mg/mL. The correlation analysis showed that the DPPH scav-
enging activity of polysaccharide fractions was associated with the content of phenolic compounds
Keywords: pectin; polysaccharides; NMR spectroscopy; arabinan; galacturonan (HG); rham-
nogalacturonan-I (RG-I); DPPH radical scavenging; phenolic compounds (PCs)
1. Introduction
Plants of the Amaranthaceae family are associated with noxious garden weeds and
ruderal plants. Perennial or annual herbaceous flowering plants of various species of the
Сhenopodium genus, known as the goosefoots, grow almost everywhere in the world and
are among the most common cosmopolitan weeds. However, this family also contains
valuable, useful plants. The genus of halophyte plant Salsola L. is one of the largest in the
family Amaranthaceae. Plants of this genus are characterized by rapid regeneration, the
ability to grow large biomass, resistance to high environmental temperature, tolerance to
soil salinity and to extended drought conditions. Therefore, the role of plants of this genus
is great in saline, arid regions of various countries with developed distant pastures. Over
150 species of the genus Salsola L., including annual semi-dwarf and dwarf shrubs and
woody trees, are distributed in arid and semi-arid regions of the Middle East, Asia, Eu-
rope and Africa [1].
Citation: Golovchenko, V.; Popov,
S.; Smirnov, V.; Khlopin, V.;
Vityazev, F.; Naranmandakh, S.;
Dmitrenok, A.; Shashkov, A.
Polysaccharides of Salsola passerina:
Extraction, Structural
Characterization and Antioxidant
Activity. Int. J. Mol. Sci. 2022, 23,
Academic Editors: Claudiu T.
Supuran and Clemente Capasso
Received: 3 October 2022
Accepted: 25 October 2022
Published: 29 October 2022
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Copyright: © 2022 by the authors. Li-
censee MDPI, Basel, Switzerland.
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tribution (CC BY) license (https://cre-
Int. J. Mol. Sci. 2022, 23, 13175 2 of 18
Extracts and decoctions of plants of this genus are used in world folk medicine to
treat bacterial and viral, cardiovascular, skin diseases, coughs and flu, and in cosmetics
[2]. Previously, several biologically active compounds were isolated from different types
of Salsola: flavonoids, phenolic acids, saponins, triterpenes, lignans, sterols, fatty acids,
alcohols, alkaloids, coumarins, as well as nitrogenous cyanogenic, isoprenoid and sul-
phur-containing compounds [3–7]. Most of these studies focus on phenolic compounds
(PCs), which attract a lot of attention because of the great antioxidant activity.
Pectiс polysaccharides are a family of complex polysaccharides present in all plant
primary cell walls [8]. The irregular block sugar chains and various macromolecular seg-
ments of the linear and ramified regions characterize the complicated structure of pectic
polysaccharides. The final model of the primary structure of the pectin macromolecule
and the model of its biosynthesis have not been developed to date. The studies spanning
the last 100 years have made it possible to establish the structure but not interlinking of
the main domains of pectins. Homogalacturonan (HG) forms linear regions and the back-
bone of the substituted galacturonans (rhamnogalacturonan II, xylogalacturonan and api-
ogalacturonan). Rhamnogalacturonan (RG) forms the backbone of the RG-I, in which the
arabinans, the galactans and/or the arabinogalactans form the side chains [9].
Pectin is resistant in the human stomach and small intestine and has to be fermented
in the large bowel by colonic bacteria. Therefore, pectin belongs to dietary fibers and pos-
sesses good prebiotic properties [10]. Pectin is highly valued as a functional food ingredi-
ent because of hypolipidemic, hypoglycemic, satiating, antibacterial and antitumor effects
[11]. In particular, pectic polysaccharides from various sources show antioxidant activity
[12–16]. Various authors suggest that galacturonic acid (GalA) scavenges free radicals
[17,18]; therefore, the HG domain mediates antioxidant activity [18,19]. Moreover, the an-
tioxidant activity of pectins may also be associated with RG-I [20]. Additionally, the side
chains of pectin may be feruloylated in certain cases, which might explain its considerable
antioxidant potential [21].
Based on the wide distribution and ability to produce a large biomass, it is of interest
to isolate pectin polysaccharides from plants of the genus Salsola L., since pectins make
up the bulk of the plant cell and are its biologically active compounds. In this study, we
assume pectins can be biologically active substances of saltwort Salsola passerine, which
remains unexplored both in terms of polysaccharide composition and low-weight molec-
ular phytochemicals and polyphenols. S. passerina, like other plant species of the genus
Salsola, grows effectively in salt soil areas and can gain high biomass in semi-arid desert
conditions, providing food for camels, sheep, goats, cattle. S. passerina is one of the most
common species in Mongolia.
The present study aims to determine the structure and antioxidant activity of poly-
saccharides isolated from the semi-shrubs of Salsola gemmascens ssp. passerina (Bunge)
Botschantz (main name—Salsola passerina Bunge) and to identify the component respon-
sible for antioxidant activity.
2. Results and Discussion
2.1. Isolation and Characterization of Polysaccharides
Five extractants—cold water, hot water, water acidified to pH 2.0, 0.7% aqueous so-
lutions of ammonium oxalate and 0.5% aqueous solutions of sodium carbonate—were
successively used to extract polysaccharides from S. passerine (Figure 1). We performed
the extraction with each extractant out until there were no sugars in the corresponding
extract. As a result, eleven polysaccharide fractions were obtained. Fractions extracted
with the sodium carbonate solution had the highest yield and those extracted with cold
water—the lowest. Polysaccharides isolated with cold water containing a significant
amount of Man, GalA and Ara residues were the principal components of polysaccharides
extracted by other extractants.
Int. J. Mol. Sci. 2022, 23, 13175 3 of 18
HW1, AC, OK1, SO1 were fractionated using a DEAE-cellulose (OH-) column. As a
result, three polysaccharide fractions were obtained from each fraction by elution with
0.01, 0.1 and 0.2 M NaCl. The polysaccharides of the obtained fractions had a similar mon-
osaccharide composition. The GalA and Ara residues were the principal components of
their carbohydrate chains. The exception was the HW1-1 fraction whose polysaccharides,
similar to the polysaccharides of fractions extracted by cold water, were characterized by
a significant content of Man and Glc residues (Table 1). This indicates that the extraction
with cold water was incomplete, and a small part of the polysaccharides was extracted
with hot water in the next step.
All parent fractions included protein components. Fractions extracted with sodium
carbonate included the largest amount of protein (up to 39%). The largest part of the pro-
teins was not connected to polysaccharides because it was removed during separation on
DEAE-cellulose. However, a small part of the co-eluted protein seemed to be connected
to polysaccharides.
Figure 1. Scheme of isolation of polysaccharides from the S. passerine.
Table 1. Chemical characteristics of pectic polysaccharides from the S. passerine.
Fractions Yield, % Content, % Mw,
kg/mol PDI
GalA c Rha
c Ara
c Xyl
c Man
c Glc
c Gal
c TS
d Protein
d PCs
CW1 0.40
a 17.46 4.33 13.72 15.45 22.45 13.82 12.77 47.44 2.3 0.8 43.59 15.62 2.79
CW2 0.14
a 31.18 5.81 8.39 10.43 24.93 7.23 12.03 46.70 6.9 1.0 63.09 12.74 4.95
CW3 0.09
a 26.14 6.92 16.13 8.12 25.35 4.67 12.67 38.33 3.7 1.0 52.68 14.04 3.75
HW1 0.88
a 47.40 4.79 11.86 9.02 11.77 7.51 7.65 64.02 3.7 0.7 96.07 27.01 3.56
HW2 0.67
a 50.44 3.62 31.59 3.26 4.12 2.63 4.35 70.27 1.9 0.7 71.44 16.02 4.46
HW1-1 6.93
b 10.12 0.59 23.12 1.03 45.41 14.07 5.65 45.00 0 nd 34.47 14.75 2.34
HW1-2 31.30
b 66.46 2.09 14.71 8.78 0.83 0.57 6.56 56.35 0 nd 56.68 20.96 2.70
HW1-3 17.40
b 79.07 4.60 4.91 5.68 0.42 0.76 4.56 76.46 0.17 nd 124.30 50.74 2.45
AC 1.82
a 47.61 3.99 31.87 6.91 2.18 1.92 5.52 52.05 2.0 0.4 55.31 13.56 4.08
AC-1 17.22
b 40.02 6.90 14.75 10.90 3.12 3.81 20.50 59.25 0 nd - - -
AC-2 50.92
b 58.46 8.28 10.50 12.83 1.51 1.64 6.77 46.92 0 nd 80.06 34.55 2.32
Int. J. Mol. Sci. 2022, 23, 13175 4 of 18
OK1 1.04
a 56.99 4.65 23.00 6.51 2.03 2.11 4.71 76.01 1.8 0.6 87.52 22.38 3.91
OK2 0.30
a 59.43 4.52 25.62 2.05 1.93 3.05 3.41 64.17 2.9 0.6 107.17 25.88 4.14
OK3 0.45
a 51.78 5.20 27.80 5.09 2.75 2.97 4.41 75.14 2.9 0.6 124.26 27.95 4.45
OK1-1 9.44
b 32.51 10.27 47.13 0.00 1.43 1.94 6.73 43.46 2.25 nd 293.36 40.02 7.33
OK1-2 25.97
b 29.00 6.00 55.24 1.41 0.18 1.46 6.71 81.05 0 nd 75.11 21.65 3.47
OK1-3 29.98
b 77.92 3.64 11.72 1.53 0.00 0.73 4.47 85.83 0 nd 92.28 44.98 2.05
SO1 7.32
a 55.48 6.84 19.42 4.31 0.00 1.83 12.12 57.84 15.6 1.2 259.19 58.09 4.46
SO2 0.12
a 52.51 7.46 24.72 1.93 0.00 3.39 10.00 62.91 18.2 1.1 239.29 30.43 7.86
SO1-1 33.78
b 21.41 8.09 60.16 1.70 0.03 0.99 7.63 88.51 10.90 0.3 444.41 319.55 1.39
SO1-2 27.97
b 33.72 4.18 47.40 3.94 0.29 1.65 8.82 77.84 0.02 0.3 93.30 33.35 2.77
SO1-3 7.88
b 60.71 6.65 19.54 5.63 0.23 0.75 6.50 68.31 5.08 0.4 81.93 40.37 2.03
a—air-dried; b—in relation to the sample applied to the column; c—data were calculated as molar %;
d—data were calculated as mass %; TS—total sugar content; PCs—phenolic compounds; nd—not
determined; number (Mn) and weight (Mw) average relative molar masses and polydispersity in-
dices (PDI) were measured using pullulan standards; the data are presented in the table as a mean
of three experiments.
2.2. NMR Spectroscopic Study
Information about the structure of the main polysaccharides from S. passerines was
obtained by a combined analysis of the NMR spectra of SO1-1, SO1-2 and SO1-3. The
NMR spectra of the three samples were similar (Figures 2–5). The 13C NMR spectra of the
samples (Figure 2) were assigned using 1H, 13C heteronuclear single quantum coherence
spectroscopy (HSQC) spectra. Analysis of the 1H, 13C HSQC spectra (Figures 3–5, Table 2)
revealed substitutions in the monosaccharide residues based on the comparison of their
13C chemical shifts with those of the parent pyranoses and furanoses [22] and considering
the glycosylation effects in the 13C NMR spectra of the carbohydrates [23,24], as well as
data from our previous NMR studies of pectins [25]. The correlated spectroscopy (COSY),
total correlation spectroscopy (TOCSY), rotating frame Overhauser effect spectroscopy
(ROESY) and heteronuclear multiple bond correlation (HMBC) spectra revealed residues
of α-D-galactopiranoside uronic acid (GA in Table 2), α-L-rhamnopyranose (R) and α-
arabinofuranose (A) in all three samples. Conclusions regarding monosaccharide compo-
sition, ring size and anomeric configuration were drawn based on the comparison of vis-
ible coupling constants and chemical shifts of the sugar residues and corresponding py-
ranoses [26,27] and furanoses [28,29].
Int. J. Mol. Sci. 2022, 23, 13175 5 of 18
Figure 2. 13C spectra of the SO1-1 (a), SO1-2 (b) and SO1-3 (c).
Figure 3. Parts of 1H, 13C HSQC spectrum of the SO1-1. The corresponding parts of the 1H and 13C
NMR spectra are shown along the X and Y axes, respectively. Arabic numerals refer to the carbon
atoms in the residues, as designated in Table 2.
Int. J. Mol. Sci. 2022, 23, 13175 6 of 18
Figure 4. Parts of 1H, 13C HSQC spectrum of the SO1-2. The corresponding parts of the 1H and 13C
NMR spectra are shown along the X and Y axes, respectively. Arabic numerals refer to the carbon
atoms in the residues, as designated in Table 2.
Figure 5. Parts of 1H, 13C HSQC spectrum of the SO1-3. The corresponding parts of the 1H and 13C
NMR spectra are shown along the X and Y axes, respectively. Arabic numerals refer to the carbon
atoms in the residues, as designated in Table 2.
Int. J. Mol. Sci. 2022, 23, 13175 7 of 18
Table 2. Chemical shifts of the signals in the 1H and 13С NMR spectra of the SO1-1, SO1-2 and SO1-
3 (323 K, D2O, TSP, δH 0.0, δC -1.6).
13С NMR Chemical Shifts (δС) and 1Н (δН, Italic), ppm
4.75 176.1
3.83; 3.73
3.89; 3.81
3.94; 3.84
TSP—trimethylsilylpropanoic acid.
The occurrence of 1,2-linked (label R) and 2,4-substituted (label R) α-L-rhamnose
residues in polysaccharides was confirmed by cross peaks at 1.25/17.9 ppm and 1.31/18.1
ppm in the 1H, 13C HSQC spectra (Figures 3–5) and HMBC spectra (Figures 6 and S2). The
ROESY spectrum of SO1-3 (Figure 7) included an inter-residue correlation peak of the
anomeric proton of Rha residues and H-4 of GalA residues at δH/H 5.26/4.44 ppm, confirm-
ing the RG-I regions in polysaccharides.
Figure 6. Part of 1H, 1H ROESY spectrum of the SO1-1. The corresponding part of the 1H NMR
spectrum is shown along the axes. Slashes refer to inter-residue correlation peaks, as designated in
Table 2.
Int. J. Mol. Sci. 2022, 23, 13175 8 of 18
Figure 7. Part of 1H, 13C HMBC spectrum of the SO1-1. The corresponding parts of the 1H and 13C
NMR spectra are shown along the X and Y axes, respectively. Arabic numerals before slash refer to
the protons, and those after slash refer to carbon atoms in the corresponding residues.
Three intense signals at δH 5.16, 5.12 and 5.09 ppm belonging to terminal nonreduc-
tion arabinose residues (label AT), 3,5-substituted arabinose residues (label AS) and 1,5-
linked arabinose residues (label AL), respectively, were found in the anomeric region of
1H NMR spectra of SO1-1 and SO1-2 (Figure S1). In the anomeric region, the 1H NMR
spectrum of SO1-3 signals of 3,5-Ara and 1,5-Ara overlaps the intense signal belonging to
the 1,4-linked D-galacturonic acid residues (label GA) at δH 5.08 ppm.
The resonance of C-6 at δC 176.0 ppm indicated the predominance of non-methyl-
esterified α-1,4-linked D-GalA residues in the structure of polysaccharides SO1-3 (Figure
2c), but the signal of low intensity at δH/C 3.86/54.4 ppm confirmed that some GalA residues
were methyl esterified.
The following inter-residue correlations H-1(glycosylating GA)/H-4(glycosylated
GA) at δH/H 5.08/4.44 ppm in the ROESY spectrum of SO1-3 (Figure 6) and H-4(glycosyl-
ated GA)/C-1(glycosylating GA) at δH/C 4.44/100.3 ppm in the HMBC spectrum of SO1-3
(Figure S2) indicated on the galacturonan (HG) in the studied polysaccharides.
The correlation peak at δН/С 2.09/21.56 ppm in the HSQC spectrum of SO1-3 (Figure
5) confirmed the O-acetylated residues in the structure of polysaccharides from S. passer-
ina. No clear evidence was obtained for the attachment of the O-acetyl group to specific
residues, since the intensity of their signals was low. Rha and GalA residues may be acetyl
esterified [30]. The signals of O-acetyl groups are present only in the spectrum of sample
SO1-3, which included polysaccharides with a high content of GalA, which may indirectly
indicate the O-acetylation of GalA residues.
The resonance of C-6 at δC 176.0 ppm indicated the predominance of non-methyl-
esterified α-1,4-linked D-GalA residues in the structure of polysaccharides (Figure 2).
The sequence of Ara residues in the repeating units was determined using the H/C
correlations in the HMBC spectra and the H/H correlations in the ROESY spectra.
The inter-residue correlation peaks—H-1(glycosylating AS)/C-5(glycosylated AS) and
H-1(glycosylating AL)/C-5(glycosylated AL) in HMBC spectra (Figures 7 and S2) and H-
Int. J. Mol. Sci. 2022, 23, 13175 9 of 18
1(AT)/H-3(AS) in ROESY spectra (Figures 6, S3 and S4)—showed the side chains formed
by single arabinose residues.
The average length of branches in the arabinan side chains, derived from the relative
amounts of terminal and branched arabinose residues, was equal to one, confirming that
the branches in the arabinan side chains comprised a single arabinose residue.
The ratio of AT, AS and AL was approximately 1:1:4 in the 1H NMR spectrum of SO1-
1 and indicated that the lengths of the linear regions were four times the lengths of the
branched regions.
The ratio of Ara residues in the spectra of SO1-2 and SO1-3 was not determined be-
cause of the overlap of the signal of the anomeric proton AL with the signal of the anomeric
proton of GA in the 1H NMR spectrum (Figure S1).
Clear inter-residue correlation peaks in the HMBC spectra (Figures 7 and S2) at δH/C
5.09/68.3 and 5.12/67.9 ppm for H-1(glycosylating AL)/C-5(glycosylated AL) and H-1(gly-
cosylating AS)/C-5(glycosylated AS) mainly indicated that 3,5-Ara substituted 3,5-Ara, and
1,5-Ara substituted 1,5-Ara.
A possible structure of the repeating unit of the arabinan chain of polysaccharides
from S. passerines is proposed below (Scheme), where the lengths of the structural regions
are arbitrary.
Scheme. A possible structure of the repeating unit of the arabinan from S. passerines.
In addition, the following low-intensity peaks were found in the anomeric region of
the 1H, 13C HSQC spectra: at δC/H 98.78/5.26 ppm belonging to Rha residues in the RG-I
regions, at δC/H 103.40/4.54, 103.80/4.49, 104.68/4.48 ppm belonging to Gal residues, respec-
tively (Figures 3 and 4).
The occurrence of 1,2-linked (label R) and 2,4-substituted (label R’) α-L-rhamnose
residues in polysaccharides was confirmed by the cross peaks at 1.25/17.9 ppm and
1.31/18.1 ppm in the 1H, 13C HSQC spectra (Figures 3–5) and the HMBC spectra (Figures
7 and S2). The ROESY spectrum of SO1-3 (Figure 6) included an inter-residue correlation
peak of the anomeric proton of Rha residues, and H-4 of the GalA residues at δH/H
5.26/4.44 ppm confirmed the RG-I regions in polysaccharides.
Thus, three structural domains were identified in the polysaccharides isolated from
S. passerine: arabinan, HG and RG-I. Considering the intensity of signals in the NMR spec-
trum, SO1-1 is dominated by the arabinan units, while SO1-3 is dominated by the galac-
turonan units. In the present study, no links between them were established. Nonetheless,
it is possible that they represent domains of a complex pectin macromolecule.
Arabinans have been found in the cell wall of several plants and are believed to form
RG-I side chains [31]. However, most of the evidence is based on co-extraction and/or co-
elution of RG-I and arabinans [32–34]. Only a few studies found that the L-Ara residues
are covalently attached to rhamnose residues at the O-4 position of the RG-I backbone
1,5-linked residues of α-L-arabinofuranose form both the backbone and the side
chains of most of the arabinans studied [37]. Backbone residues are usually substituted at
O-2 and/or O-3 and/or at both positions, with O-3 substitutions predominating [32]. How-
ever, several arabinans with a high percentage of substitution at the O-2 position have
also been found [38,39]. Other structures of arabinans have also been described. For ex-
ample, in arabinans, both the furanose and pyranose forms of arabinose were found [40].
Int. J. Mol. Sci. 2022, 23, 13175 10 of 18
Terminal β-arabinofuranose residues may glycosylate 1,5-linked α-arabinofuranose resi-
dues of the backbone at position O-5 [41]. Various degrees of branching have been found,
including single, linear and branched oligomeric and polymeric chains, with different
linkage types. The almost linear 1,5-arabinan associated with protein was isolated from
red wine [42]. The arabinans in pectins often have single substituted side chains [32,43].
Arabinans from soybean [44], apple [45], the inner bark of Norway spruce [46] were found
to have a highly branched structure. Arabinan-rich pectins, which constituted 50% of the
total pectic polysaccharides, have been obtained from pea Pisum sativum L. [47].
The roles of arabinans in plant cell walls remain unclear. It was established that arabi-
nans can be substituted by terminal phenolic esters, particularly feruloyl or coumaroyl
esters. Ferulic acid groups may be ester linked to O-2 of the arabinose residues [48,49].
Feruloyl esters may determine guard cell wall flexibility by providing the cross-links be-
tween arabinans and other wall polymers; this testifies a unique role for arabinans in de-
termining the physical and functional properties of guard cell walls [50].
2.3. DPPH Radical-Scavenging Activity
Polysaccharide fractions from S. passerine scavenged the DPPH radical at concentra-
tions of 0.2–1.8 mg/mL. The half-maximal DPPH inhibitory concentration (IC50) of them is
given in Table 3. CW2, CW3, SO1 and SO2 demonstrated the highest activity, which ex-
ceeded 2.51–2.96 times that of commercial apple pectin (AP) activity. CW2, CW3, SO1 and
SO2 scavenged 67, 69, 55 and 67% of DPPH radicals at a concentration of 1 mg/mL. Other
fractions were less effective and scavenged only 31–48% of DPPH radicals at a concentra-
tion of 1 mg/mL.
Table 3. DPPH scavenging effect of polysaccharide fractions from S. passerine.
Pectin IC50 (mg/mL)
CW1 1.14 ± 0.02 cd
CW2 0.69 ± 0.07 bc
CW3 0.66 ± 0.07 b
HW1 1.47 ± 0.22 d
HW2 1.58 ± 0.24 d
AC1 2.20 ± 0.41 e
OK1 1.63 ± 0.26 d
OK2 1.84 ± 0.03 de
OK3 1.64 ± 0.11 d
SO1 0.78 ± 0.06 bc
SO2 0.68 ± 0.08 b
AP 1.96 ± 0.36 de
Trolox 0.006 ± 0.001 a
Data are presented as the mean ± SD of three independent experiments. Different capital letters (a–
e) show the significant differences (p < 0.05, LSD test).
The DPPH radical scavenging assay is widely used to evaluate the antioxidant prop-
erty of plant polysaccharides. The activity of CW2, CW3, SO1 and SO2 seems to be com-
parable to that of polysaccharides from cantaloupe rinds [51], hawthorn wine pomace
[52], fruit bodies of Tremella fuciformis [53] and apple pomace [54]. It should be noted that
some other polysaccharides demonstrated the same level of DPPH scavenging activity at
lower concentrations. These include pectins from Chaenomeles sinensis fruits [55], Epilobium
angustifolium L. [56], Thymus quinquecostatus Celak. leaves [57], Gardenia jasminoides J. Ellis
flowers [58] and Ziziphus jujuba cv. Muzao [59].
The DPPH radical scavenging activity of the fractions obtained by DEAE-cellulose
elution was compared with the activity of the parent fraction SO1 (Figure 8). The polysac-
charides SO1-1, SO1-2 and SO1-3 obtained were less effective (p < 0.05) than the parent
Int. J. Mol. Sci. 2022, 23, 13175 11 of 18
fraction SO1, exhibiting IC50 equal to 3.64 ± 0.18, 5.70 ± 0.92 and 5.70 ± 1.30 mg/mL, respec-
Figure 8. The DPPH radical scavenging activity of SO1 and of its components obtained by separa-
tion on DEAE-cellulose column. Data are presented as the mean ± SD of three independent experi-
On the basis of the yield and content of PCs in SO1-1, SO1-2, SO1-3 and SO1, most
of the PCs providing antioxidant activity were removed by anion exchange chromatog-
raphy, assuming that they were not bound to the polysaccharide chains. The sum contents
of PCs in polysaccharides SO1-1, SO1-2, SO1-3 included about 18% from the content of
PCs in the parent fraction SO1. It was detected that three fractions obtained on DEAE-
cellulose provided only 24% of the DPPH radical scavenging activity of SO1, although
they represented about 70% of the parent pectin (Table 1). This suggests that the antioxi-
dant activity of SO1 was mainly provided by the associated PCs but not by polysaccha-
The relationship between the chemical characteristics of polysaccharides and DPPH
scavenging ability was further investigated using correlation analysis. The total content
of sugars and the (Ara + Gal)/Xyl ratio correlated negatively, whereas the content of PCs,
Gal and Man, as well as PDI correlated positively with DPPH scavenging activity (Table
Table 4. The Pearson correlation coefficients between the DPPH scavenging activity and the chem-
ical characteristics of polysaccharide fractions from S. passerine (n = 14).
Second Variable R p Second Variable R p
PCs 0.89 0.000 Protein 0.51 0.062
Gal 0.59 0.026 Glc 0.46 0.098
PDI 0.58 0.029 Rha 0.34 0.236
Man 0.55 0.040 Xyl/GalA 0.30 0.290
(Ara + Gal)/Rha 0.58 0.028 Xyl 0.27 0.342
Total sugar 0.67 0.009 Rha/GalA 0.28 0.335
Mw 0.11 0.713
Mn 0.12 0.671
Ara/Xyl 0.22 0.455
GalA 0.23 0.430
GalA/NM 0.25 0.384
Int. J. Mol. Sci. 2022, 23, 13175 12 of 18
GalA-Rha 0.26 0.377
2Rha + Ara + Gal 0.30 0.295
Ara 0.52 0.059
We tested the five regression models, subsequently removing the less significant fac-
tors (according to the p-value). The linear regression, including the contents of PCs and
Man as independent variables, resulted in the best model for prediction (adj. R2 = 0.82, p =
0.000) (Table 5). The content of PCs was the only factor contributing significantly to DPPH
scavenging activity (p = 0.000, β = 0.79).
Table 5. Multiple regression analysis of the DPPH scavenging activity.
Variable β Standard
Error of β
Error p-Value
Dependent Independent
DPPH scavenging
PCs 0.79 0.13 2.02 0.32 0.000
Man 0.26 0.13 0.02 0.01 0.062
Regression results: R2 = 0.847, adjusted R2 = 0.819, F2.11 = 30.438, p < 0.000, Standard estimate error =
Thus, the correlation analysis showed that the DPPH scavenging activity of the sam-
ple from S. passerine is associated with the content of PCs. This is consistent with the re-
sults of Ref [54], whose authors evaluated the activity of apple pectins, and our previous
study on fireweed pectins [56]. It is known that PCs may bind covalently to the side chains
of RG I through the Ara and Gal residues and may be involved in the cross-linking of
macromolecules [60].
It was shown that the removal of PCs from polysaccharides reduces the antioxidant
activity but does not completely abolish it [60]. Several authors suggest that the antioxi-
dant activity of pectins may be due to the hydroxyl and carboxyl groups of GalA residues
[52,61]. Previously, we showed that the antioxidant activity of fireweed pectins is partly
related to the xylogalacturonan chains [56]. However, in the present study, we failed to
find the polysaccharide chains responsible for the DPPH radical scavenging activity of
Salsola pectins. The small sample size (n = 14), which determines the statistical power of
multiple regression [62], may be the reason we failed to identify the polysaccharide chains
that contribute to the antioxidant activity of Salsola pectins.
3. Materials and Methods
3.1. Materials
Biological material: plant material, consisting of yellow-green annual branches with
spherical dwarf leaves, was collected in August 2019 from the semi-shrubs of Salsola gem-
mascens ssp. passerina (Bunge) Botschantz. = Caroxylon passerinum (Bunge) Akhani et E.H.
Roalson (main name—Salsola passerina Bunge) growing in Mandal-ovoo soum, Ömnö-
Govi province, Mongolia. They were identified by Prof. B.Oyuntsetseg (School of Arts and
Sciences, National University of Mongolia). The plant material was washed with distilled
water and dried with filter paper.
The chemicals used are described in the Supplementary (Appendix A).
3.2. Isolation of Polysaccharides of S. passerina
Polysaccharides from the plant material were sequentially extracted, as described be-
low; the extraction scheme is shown in Figure 1. At each stage, an exhaustive extraction
of polysaccharides was carried out until the absence of reaction of the extract to the car-
bohydrate; the extraction mixtures were mixed in a mechanical stirrer.
Freshly picked plant material (234 g) was milled in a blender, distilled water (1 L)
was added, and the resulting mixture was stirred in a mechanical mixer at 20 °C for 3 h.
The mixture was centrifuged, and the residue of the plant material was treated again; the
Int. J. Mol. Sci. 2022, 23, 13175 13 of 18
treatment was repeated three times. In the next stage, polysaccharides from the residues
of plant materials were extracted with hot water at 80 °C for 3 h. The extraction was re-
peated twice (each time, the volume of added water was 1 L). Finally, the five aqueous
extracts (three obtained with cold water (CW1, CW2, CW3) and two with hot water (HW1,
HW2)) were obtained. Next, polysaccharides were extracted with acidified water (pH 2.0,
1 L) at 50 °C for 3 h. As a result, one extract (AC) was obtained. Next, polysaccharides
were extracted with aqueous solutions of ammonium oxalate (0.7% w/v) at 70 °C for 6 h.
The extraction was repeated three times (the first volume of salt solution added was 2 L;
the second and third volumes were 1 L). Finally, the three extracts (OK1, OK2, OK3) were
obtained. Next, polysaccharides were extracted with aqueous solutions of Na2CO3 (0.5%
w/v) containing NaBH4 at 70 °C for 3 h. The extraction was repeated twice (the first volume
of soda solution added was 3 L, the second—2 L). The two extracts (SO1, SO2) were ob-
The carbohydrate content of each extract was detected using a phenol-sulfuric acid
assay [63].
All extracts were dialyzed against distilled water for 48 h at 10 °C. Extracts SO1, SO2
were previously acidified with a diluted solution of acetic acid to pH 5.6. The dialyzed
extracts were concentrated on a Heidolph 4002 rotary evaporator (Germany) under re-
duced pressure at 40 °C.
Polysaccharides were precipitated from the extracts with a four-fold volume of 95%
ethanol, centrifuged, washed twice with 95% ethanol, dissolved in distilled water, frozen
and lyophilized. The yields of the polysaccharide fractions obtained are expressed in %
(w/w) of mass of dry plant material and are presented in Table 1.
3.3. Ion Exchange Chromatography of Polysaccharide Fractions
The major polysaccharide fractions HW1, AC, OK1, SO1 were separated on a DEAE-
cellulose (OH-) column (2.5 cm × 40 cm). Each polysaccharide fraction (100 mg) was dis-
solved in 5 mL of 0.01 M NaCl, and the solution was applied to the column. The column
was stepwise eluted with 0.01, 0.1, 0.2, 0.3, 0.5 and 1.0 M NaCl solution (400 mL of each
eluent) at a flow rate of 0.9 mL/min. The fractions were collected at 12 min intervals using
a low-pressure system Pharmacia Biotech (Sweden) with a FRAC-100 fraction collector,
P-50 pump. The carbohydrate content in each tube was determined by the phenol–sulfuric
acid method [63]. When separating each of the HW1, OK1, SO1, three major polysaccha-
ride fractions were obtained (eluted with 0.01, 0.1 and 0.2 M NaCl). When separating AC,
the fraction eluted 0.2 M NaCl was obtained as minor. In addition, minor fractions were
obtained from all fractions by elution with 0.3, 0.5 and 1.0 M NaCl.
The separation procedure was repeated twice for HW1, OK1 and four times for AC,
SO1. Data on the monosaccharide composition and the yield of the fractions are presented
in Table 1 as a mean of these experiments.
3.4. General Analytical Methods
The content of uronic acids was determined as described earlier [64,65]. The quanti-
tative determination of protein was calculated using the Bradford method [66]. The quan-
titative determination of phenolics was performed with the Folin–Ciocalteu reagent using
gallic acid as a standard [67]. The content of neutral monosaccharides was determined by
gas–liquid chromatography (GLC), as described earlier in detail [68]. The sugar concen-
tration was determined at 490 nm using the phenol–sulfuric acid assay [63].
The relative molar mass distributions (RMM) (including Mn, Mw and PDI) of the
polysaccharide samples were determined by size exclusion chromatography with high-
performance liquid chromatography (HPSEC); the procedure was described in detail ear-
lier [69].
Int. J. Mol. Sci. 2022, 23, 13175 14 of 18
3.5. Nuclear Magnetic Resonance Spectroscopy
All homo- and heteronuclear NMR experiments of the samples were carried out on
a Bruker Avance 600 spectrometer (Germany) at a probe temperature of 303, 313 and 318
K, which provided a minimum overlap of the signal of deuterated water with the polymer
signals. The procedures for preparing the polysaccharide samples and the conditions of
the NMR experiments were described earlier [69].
3.6. Antioxidant Activity
The DPPH solution (0.2 mM, in ethanol) was added to the pectin solution (0.4–3.6
mg/mL water) in equal proportions (v/v) and mixed. After incubation at 25 °C for 1 h, the
absorbance of the sample was measured at 517 nm. The scavenging activity of the pectins
was measured at four different concentrations, and the half-maximal inhibitory concen-
tration (IC50, mg/mL) values were calculated based on a polynomial regression curve [70].
3.7. Statistical Analysis
The significance of the difference among the means in determining the antioxidant
activity was estimated with one-way analysis of variance (ANOVA) and Fisher’s least sig-
nificant difference (LSD) post hoc test at p < 0.05. The relationship between the chemical
characteristics and activity of polysaccharide fractions was evaluated by the calculation
of the Pearson correlation coefficients and multiple linear regression analysis. All calcula-
tions were performed using the statistical package Statistica 10.0 (StatSoft, Inc., USA). The
data were expressed as the means ± s.d. of three independent experiments.
4. Conclusions
Polysaccharide fractions isolated from S. passerine with water and aqueous solutions
of ammonium oxalate and sodium carbonate were characterized by a similar composition,
including polysaccharides, protein and PCs. HG, RG-I and arabinan with regions formed
by 3,5-substituted and by 1,5-linked arabinose residues were identified as the principal
units of the polysaccharides obtained. Polysaccharide fractions of S. passerine demon-
strated a moderate antioxidant potential. Fractions isolated with cold water and sodium
carbonate scavenged the DPPH radical in vitro to a much greater extent than commercial
apple pectin. The correlation analysis of the composition and activity of polysaccharide
fractions obtained by anionic exchange chromatography revealed that the antioxidant ca-
pacity of polysaccharides of S. passerine is mainly due to the associated PCs.
Supplementary Materials: The following supporting information can be downloaded at:
Author Contributions: Study design, conceptualization, polysaccharide isolation, V.G.; writing—
review and editing, project administration, S.P.; measurement of antioxidant activity, V.S.; separa-
tion of polysaccharide fractions on an anion exchange resin, V.K.; collecting plant material and its
primary treatment by organic solvents, F.V.; collecting plant material and its primary treatment by
organic solvents, S.N.; experiments and analysis of NMR data, A.S.S. and A.S.D. All authors have
read and agreed to the published version of the manuscript.
Funding: This study was supported by the Russian Science Foundation (grant number 21-73-20005).
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data that support the findings of this study are available from the
corresponding author upon reasonable request.
Acknowledgments: NMR spectra were registered on the equipment of the Zelinsky Institute of Or-
ganic Chemistry Shared Research Facility.
Conflicts of Interest: The authors declare no conflicts of interest.
Int. J. Mol. Sci. 2022, 23, 13175 15 of 18
Appendix A
Chemical reagents: ethyl alcohol, ethanol, C2H5OH (96%, JSC Kirov Pharmaceutical
Factory, Russia); methyl alcohol, methanol, CH3OH (99.9%, Reakhim, Russia); sodium hy-
droxide, NaOH (98%, Fluka, Germany); sodium chloride, NaCl (99%, Sigma-Aldrich,
USA); chloroform, CHCl3 (>99.9%, Ekos-1, Russia); ammonium hydroxide solution,
NH4OH ((25 NH3 in H2O, >99.9%, Ekos-1, Russia); trifluoroacetic acid, CF3COOH (99%,
Acros organics, USA); pyridine, C5H5N (99%, Ekos-1, Russia); acetic acid, CH3COOH
(99.9%, Khimreactive, Russia); sodium borohydride, NaBH4 (>98.5%, Sigma-Aldrich,
USA); D2O (99.9 atom % D, Sigma-Aldrich, USA); toluene, C6H5-CH3 (99%, Ekos-1, Rus-
sia); sulfuric acid, H2SO4 (>99.9%, Vekton, Russia); phenol, C6H5OH (99%, Reakhim, Rus-
sia); 3,5-dimethylphenol, (CH3)2C6H3OH (99%, Sigma-Aldrich, USA); 1,4-α-D-polygalac-
turonase (Sigma, exo- and endo-activity 690 units/g); 1,1-diphenyl-2-picrylhydrazyl radi-
cal (Sigma-Aldrich, USA); the Folin and Ciocalteu’s phenol reagent (Sigma-Aldrich, USA).
For the ion exchange chromatography, we used DEAE-cellulose (Sigma-Aldrich,
The bovine serum albumin (96%, Sigma-Aldrich, USA); myo-inositol (99%, Sigma-
Aldrich, USA), L-(+)-Rhap, L-(+)-Araf, D-(+)-Galp, D-(+)-Manp, D-(+)-xylose and D-(+)-Glcp
(99%, Sigma-Aldrich, USA); D-(+)-GalpA monohydrate (97%, Sigma-Aldrich, USA); gal-
lic acid (MP Biomedicals, USA) were used as standards.
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... Equal concentrations of SCCP were prepared and measured in the same manner; the same concentration of ascorbic acid served as a positive control [52]. ...
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This study aimed to enhance the utilization value of sweet corn cob, an agricultural cereal byproduct. Sweet corn cob polysaccharide-ron (III) complexes were prepared at four different temperatures (40 °C, 50 °C, 60 °C, and 70 °C). It was demonstrated that the complexes prepared at different temperatures were successfully bound to iron (III), and there was no significant difference in chemical composition; and SCCP-Fe-C demonstrated the highest iron content. The structural characterization suggested that sweet corn cob polysaccharide (SCCP) formed stable β-FeOOH iron nuclei with −OH and −OOH. All the four complexes’ thermal stability was enhanced, especially in SCCP-Fe-C. In vitro iron (III) release experiments revealed that all four complexes were rapidly released and acted as iron (III) supplements. Moreover, in vitro antioxidant, α-glucosidase, and α-amylase inhibition studies revealed that the biological activities of all four complexes were enhanced compared with those of SCCP. SCCP-Fe-B and SCCP-Fe-C exhibited the highest in vitro antioxidant, α-glucosidase, and α-amylase inhibition abilities. This study will suggest using sweet corn cobs, a natural agricultural cereal byproduct, in functional foods. Furthermore, we proposed that the complexes prepared from agricultural byproducts can be used as a potential iron supplement.
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The genus Salsola L. (Russian thistle, Saltwort) includes halophyte plants and is considered one of the largest genera in the family Amaranthaceae. The genus involves annual semi-dwarf to dwarf shrubs and woody tree. The genus Salsola is frequently overlooked, and few people are aware of its significance. The majority of studies focus on pollen morphology and species identification. Salsola has had little research on its phytochemical makeup or biological effects. Therefore, we present this review to cover all aspects of genus Salsola, including taxonomy, distribution, differences in the chemical constituents and representative examples of isolated compounds produced by various species of genus Salsola and in relation to their several reported biological activities for use in folk medicine worldwide.
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The aim of this study was to isolate pectins with antioxidant activity from the leaves of Epilobium angustifolium L. Two pectins, EA-4.0 and EA-0.8, with galacturonic acid contents of 88 and 91% were isolated from the leaves of E. angustifolium L. by the treatment of plant raw materials with aqueous hydrochloric acid at pH 4.0 and 0.8, respectively. EA-4.0 and EA-0.8 were found to scavenge the DPPH radical in a concentration-dependent manner at 17–133 μg/mL, whereas commercial apple pectin scavenged at 0.5–2 mg/mL. The antioxidant activity of EA-4.0 was the highest and exceeded the activity of EA-0.8 and a commercial apple pectin by 2 and 39 times (IC50—0.050, 0.109 and 1.961 mg/mL), respectively. Pectins EA-4.0 and EA-0.8 were found to possess superoxide radical scavenging activity, with IC50s equal to 0.27 and 0.97 mg/mL, respectively. Correlation analysis of the composition and activity of 32 polysaccharide fractions obtained by enzyme hydrolysis and anionic exchange chromatography revealed that the antioxidant capacity of fireweed pectins is mainly due to phenolics and is partially associated with xylogalacturonan chains. The data obtained demonstrate that pectic polysaccharides appeared to be bioactive components of fireweed leaves with high antioxidant activity, which depend on pH at their extraction.
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Pectin is a kind of natural and complex carbohydrates which is extensively used in food, chemical, cosmetic, and pharmaceutical industries. Fresh sunflower ( Helianthus annuus L.) heads were utilized as a novel source of pectin extracted by ammonium oxalate. The conditions of the extraction process were optimized implementing the response surface methodology. Under optimal extraction parameters (extraction time 1.34 h, liquid–solid ratio 15:1 mL/g, ammonium oxalate concentration 0.76% (w/v)), the maximum experimental yield was 7.36%. The effect of spray-drying and freeze-drying on the physiochemical properties, structural characteristics, and antioxidant activities was investigated by FT-IR spectroscopy, high performance size exclusion chromatography, and X-ray diffraction. The results showed freeze-drying lead to decrease in galacturonic acid (GalA) content (76.2%), molecular weight ( M w 316 kDa), and crystallinity. The antioxidant activities of pectin were investigated utilizing the in-vitro DPPH and ABTS radical-scavenging systems. This study provided a novel and efficient extraction method of sunflower pectin, and confirmed that different drying processes had an effect on the structure and properties of pectin.
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The biological activity of apple pectin extracted conventionally or enzymatically using endo-xylanase and endo-cellulase, was tested in vitro. The analyses were performerd in tetraplicates and the statistical significance of the differences were assessed using ANOVA, Tukey post hoc and LSD (the least significant difference) tests. Multivariate regression analysis was applied to determine the structural components that have a crucial importance for antioxidant and antitumor properties of pectins. The pectins extracted by enzymes contained up to four times more ferulic acid and showed twice as great ability to neutralize free radicals and Fe(III) reduction. The antiradical potential positively correlated with phenols, fucose and rhamnose content. In the assays performed on HT-29 human adenocarcinoma and B16F10 melanoma cell cultures, the “green” pectins, contrary to acid isolated ones, exhibited remarkable anti-neoplastic potential while being nontoxic to nontransformed L929 cell line. The pectins in the dose of 1 mg/mL were capable of inhibiting adhesion (max 23.1%), proliferation (max 40.4%), invasion (max 76.9%) and anchorage-independent growth (max 90%) of HT-29 cells (significance level p < 0.001). These pectin preparations were slightly less active towards B16F10 cells. The enzyme-isolated apple pectins may be useful as a functional food additive and an ingredient of the ointment formulas for post-surgical melanoma treatment.
We report that in birch leaf pectin, rhamnogalacturonan-I (RG-I) and galacturonan (HG) were found as separate polymers rather than domains of a complex macromolecule. RG-I and HG were separated by anion-exchange and size-exclusion chromatography and studied by using NMR spectroscopy. NMR spectra showed that methyl-esterified D-galactosyluronic acid residues were located only in HG. Oligosaccharides of similar structure to the backbone, but without terminal reducing residues in the NMR spectra, were found in RG-I. We hypothesize, these oligosaccharides and RG-I backbone can be covalently bound due to its co-eluted of from DEAE-cellulose and Sepharose CL-4B. This result differs from the classical RG-I model, which assumes that all Rhap and GalpA residues are located only in the RG-I backbone. In the heteronuclear multiple bond correlation (HMBC) and rotating frame Overhauser effect spectroscopy (ROESY) spectra, the correlation peaks confirming the substitution of 2,4-rhamnose residues at O-4 by only single D-galactose residues were identified.
In this study, two acidic Biluochun Tea polysaccharides (BTP-A11 and BTP-A12) were investigated comparatively, which mainly consisted of Rha, Ara, Gal and GalA, possibly suggesting their pectic nature. Structurally, their galacturonan backbones composed of →4)-α-D-GalpA-(1→ and →2)-α-L-Rhap-(1→ were revealed similar, while Ara- and Gal-based branches attached to the O-2 of →2)-α-L-Rhap-(1→ were in distinctive types, proportions, extensibilities and branching degrees. This could lead to their different macromolecular characteristics, where BTP-A11 with higher Mw presented a more hyper-branched chain conformation and relatively higher structural flexibility/compactness, thereby resulting in a lower exclusion effect and an insufficient hydrodynamic volume. Besides, better radical scavenging activities in vitro were also determined for Gal-enriched BTP-A11, where a larger surface area containing more H-donating groups were related to its higher Mw, more hyper-branched conformation, lower DM and higher DA. Therefore, the understanding of structure-property-activity relationships was improved to some degrees for acidic Biluochun Tea polysaccharides, which could be potentially required for more applications in food, medical and cosmetic fields.
In this study, polysaccharides (DJPs) were obtained from Thymus quinquecostatus Celak. leaves, which were DJP (the crude polysaccharide extracted by hot water), DJP30, DJP50, DJP70 (three polysaccharide fractions obtained by graded precipitation with final ethanol concentrations of 30%, 50%, and 70%), and DJP70-1 (a purified polysaccharide isolated from DJP70 by DEAE-52 cellulose column). DJPs were all high-molecular-weight heteropolysaccharides, with different contents of total carbohydrate, protein, uronic acid, and phenols. They were mainly composed of arabinose, galactose, glucose, galacturonic acid, and rhamnose, and DJP70-1 also surprisingly contained a higher proportion of mannose (Man). Structural characteristics were determined using infrared spectrum (IR), methylation-gas chromatography-mass spectrometry (GC-MS), and 1D/2D nuclear magnetic resonance (NMR) analysis, with the existence of 14 kinds of glycosyl linkages of DJP70-1. Morphological features of DJP70-1 determined by Congo red and scanning electron microscopy (SEM) were also significantly different from other DJPs. Moreover, these polysaccharides exhibited strong scavenging effects on 2, 2′-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals, though the relatively weak scavenging effect on hydroxyl free radicals in vitro. Furthermore, the crude polysaccharide DJP exerted a protective effect against 2, 2′-azo-bis-(2-methylpropylimid)-dihydrochloride (AAPH)-induced oxidative stress in the zebrafish model by directly scavenging excessive reactive oxygen species (ROS) and activating the kelch-like ECH-associated protein 1 (KEAP1) - nuclear factor erythroid 2-related factor 2 (NRF2)/antioxidant response element (ARE) signaling pathway in vivo. α-amylase and α-glucosidase inhibitory activities evaluation results showed that DJPs were also good potential hypoglycemic agents.
Background: The rinds from cantaloupe is an agricultural waste of cantaloupe industrial processing. The current study tried to (i) evaluate the potential use of cantaloupe rind as a pectin source, (ii) optimize the factors of microwave-assisted extraction process using Box-Behnken design, and (iii) characterize the isolated pectin using various physicochemical, structural, functional, and bioactivity properties. Results: Four variables of the extraction process were successfully optimized at microwave power of 700 W, irradiation time of 112 s, pH value of 1.50, and LS value: 30 mL/g with a yield of 181.4 g kg-1 . The analysis indicated a high-methylated galacturonic acid-rich (703.4 g kg-1 ) sample with an average molecular weight of 390.475 kDa. Also, the isolated pectin showed considerable functionality and antioxidant ability. The main functional groups, structural characteristics, and crystallinity of samples were comparatively studied using FTIR, NMR and XRD spectroscopies. Conclusion: In comparison to commercial citrus pectin, isolated pectin showed a significantly higher value for most of the functional analysis such as oil holding capacity, emulsifying capacity, emulsion stability, DPPH• and ABTS•+ scavenging activity, and reducing power assay. In other analyses the isolated sample was close to the commercial one, meaning that the cantaloupe rinds should be considered as a suitable additional resource for pectin production. This article is protected by copyright. All rights reserved.
Two pectic polysaccharides (WRSP-A2b and WRSP-A3a) have been obtained from Radix Sophorae Tonkinensis and comparatively investigated in terms of their physical properties and antioxidant activities. Monosaccharide composition, FT-IR, NMR and enzymatic analyses indicate that both WRSP-A2b (13.6 kDa) and WRSP-A3a (44.6 kDa) consist of homogalacturonan (HG), rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II) domains, with mass ratios of 0.9:1.8:1 and 2.3:2.9:1, respectively. The RG-I domains were further purified and characterized. Results show that WRSP-A2b contains a highly branched RG-I domain, primarily substituted with α-(1→5)-linked arabinans, whereas WRSP-A3a contains a small branched RG-I domain mainly composed of β-(1→4)-linked galactan side chains. WRSP-A3a exhibits stronger antioxidant activity in scavenging different radicals than WRSP-A2b, a finding that may be due to its higher content of GalA residues and HG domains. Our results provide useful information for screening natural polysaccharide-based antioxidants from Radix Sophorae Tonkinensis.
The effects of extraction conditions on the yield of polysaccharides from the fruit of Chaenomeles sinensis (FCS) using a hot compressed water method were investigated. The results showed that an appropriately high temperature (150 °C) and a moderate extraction time (45 min) at a material to water ratio of 1 to 10 g/mL led to a high yield of alcohol precipitation polysaccharide (PA). The purified polysaccharides (CSP-1, CSP-2, and CSP-3) were successfully obtained using a DEAE-52 chromatographic column. Chemical analysis showed that CSP-2 and CSP-3 were homogenous and exhibited characteristics of esterified pectins, whereas CSP-2 mainly consisted of galacturonic acid (GalA), galactose (Gal), arabinose (Ara), rhamnose (Rha), and mannose (Man) with an average molecular weight of 59.1 kDa. Furthermore, CSP-1 possessed stronger antioxidant ability according to DPPH scavenging and reducing power compared with CSP-2 and CSP-3. However, it was weaker with respect to OH scavenging. The technical data presented in this study could help the industry make use of polysaccharides from FCS as a source of pectin for a range of pharmaceutical, culinary, and cosmetic products.