Antioxidant, antiradical, and antimicrobial activities of polysaccharides obtained by microwave-assisted extraction method: A review

Journal Pre-proof Antioxidant, antiradical, and antimicrobial activities of polysaccharides obtained by microwave-assisted extraction method: A review Monirsadat Mirzadeh, Mohammad Reza Arianejad, Leila Khedmat

PII:

S0144-8617(19)31088-4

DOI:

https://doi.org/10.1016/j.carbpol.2019.115421

Reference:

CARP 115421

To appear in:

Carbohydrate Polymers

Received Date:

8 August 2019

Revised Date:

22 September 2019

Accepted Date:

30 September 2019

Please cite this article as: Mirzadeh M, Arianejad MR, Khedmat L, Antioxidant, antiradical, and antimicrobial activities of polysaccharides obtained by microwave-assisted extraction method: A review, Carbohydrate Polymers (2019), doi: https://doi.org/10.1016/j.carbpol.2019.115421

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Antioxidant, antiradical, and antimicrobial activities of polysaccharides obtained by microwave-assisted extraction method: A review Monirsadat Mirzadeh1, Mohammad Reza Arianejad2, Leila Khedmat3, * 1

Metabolic Disease Research Center, Qazvin University of Medical Sciences, Qazvin, Iran

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Health Management Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

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Department of Food Science and Technology, Faculty of Nutrition, Tabriz University of Medical Sciences, Tabriz, Iran

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*Corresponding author’s Email: [email protected]; Tel: +98-912-387-3622

Highlights

Great bio-functionalities of polysaccharides extracted by microwave-assisted (MAE)

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The high capacity of polysaccharides to quench DPPH·, OH·, NO·, ABTS·+, and O2·−

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The dose-dependent reducing, chelating, and lipid peroxidation inhibition activities

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Uronic acids are the main constituents involved in the antioxidative properties

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Strong antibacterial, antifungal, and antiviral activities of MAE-polysaccharides

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Abstract

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The antioxidant and antimicrobial activities of polysaccharides extracted by microwave-assisted extraction (MAE) were reviewed. An ascending dose-dependent manner was found for the in vitro antioxidant (e.g., nitrite scavenging, phospho-molybdenum reduction, inhibition of lipid

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peroxidation (ILP), ferric reducing power, and ferrous metal ions chelating), and antiradical (against DPPH·, OH·, ABTS·, NO·, and O2·−) activities. There was a positive and significant correlation between ILP and erythrocyte hemolysis inhibition, showing the excellent antioxidative properties to prevent the risk of cell damage. These carbohydrate-based polymers in vivo could reduce malonaldehyde and protein carbonyls and increase stress-resistance-related enzymes such as catalase, superoxide dismutase, and glutathione peroxidase. They showed an effective bactericidal activity against a wide variety of gram-negative and gram-positive bacterial 1

infections. The in vitro strong antifungal and antiviral activities of sulfated polysaccharides extracted by MAE were also diagnosed without any cytotoxicity effect. Therefore, these biomacromolecules might be used to develop functional foods and nutraceuticals.

Abbreviations UAE: ultrasound-assisted extraction, MAE: microwave-assisted extraction, PWE: pressurized water extraction, SWE: subcritical water extraction, EAE: enzyme assistance extraction, HWE:

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hot-water extraction, SFE: supercritical fluid extraction, Mw: molecular weight, PMRA: phospho-molybdenum reduction activity, ILP: inhibition of lipid peroxidation, FRAP: ferric

reducing antioxidant power, FICA: ferrous ion chelating activity, RP: reducing power, ROSs:

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reactive oxygen species, AAPH: 2,2′‐Azobis(2‐amidinopropane) dihydrochloride, ORAC:

oxygen radical absorbance capacity, PCCs: positive control compounds, MVI: methyl viologen

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induction, HPI: hydrogen peroxide induction, MDA: malonaldehyde, SOD: superoxide dismutase, PCO: protein carbonyls, CAT: catalase, GSH: glutathione, GSH-Px: glutathione

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peroxidase, NO: nitric oxide, iNOS: nitric oxide synthase, CYP: cyclophosphamide, ST: survival time, RBP: rice bran polysaccharide, MIC: minimum inhibitory concentration, RSM: response

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surface methodology.

Keywords: Bioactive polysaccharide, Extraction, Microwave, Antioxidant, Antibacterial,

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Disease prevention

1. Introduction

Polysaccharides are biopolymers consisting of a high number of monosaccharides linked by glycosidic bonds. These macromolecules have been utilized as non-toxic ingredients to design

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functional products in agriculture, health-food, cosmetics, and medical industries (Chen, Ji, Xu, & Liu, 2019; Hu, Liu, et al., 2019). The various extraction methods are applied to extract polysaccharides from natural sources such as plant and algae materials, and microorganisms. The extraction technique plays a significant role in the yield, chemical structure, and bioactivity of carbohydrate-based polymers (Guo et al., 2019; Rostami & Gharibzahedi, 2017a, b). Therefore, it is necessary to find an appropriate extraction method to maintain the physicochemical,

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rheological, functional, and structural properties of polysaccharides (Cheng, Feng, Jia, Li, Zhou, & Ding, 2013; Gharibzahedi, Smith, & Guo, 2019a; Yuan, Xu, Jing, Zou, Zhang, & Li, 2018). A hot reflux unit during a multiple-step process is applied to extract polysaccharides in the conventional method. This traditional process under high temperatures usually leads to the low extraction rate of polysaccharides at extended times (Hu, Zhao, et al., 2019; Tahmouzi, 2014). However, the yield and bioactivity of polysaccharides can be promoted by some high-efficient techniques such as ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE),

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pressurized water extraction (PWE), and enzyme assistance extraction (EAE) (Chen, Fang, Ran, Tan, Yu, & Kan, 2019; Gharibzahedi & Mohammadnabi, 2017; Li, Wang, Zhang, Huang, & Ma, 2011). MAE is an innovative extraction system to extract high-yield polysaccharides at a shorter

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time and lower solvent and energy consumption (Fig. 1). This extraction method has higher

extraction rate compared to the other technologies such as Soxhlet, PWE, and supercritical fluid

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extraction (SFE) owing to the rapid heating (Zeng, Zhang, Gao, Jia, & Chen, 2012). Also, MAE as a sample preparation technique could be easily coupled with analytical systems such as

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chromatography and spectroscopy to assess bioactive compounds (Xia, Wang, Xu, Zhu, Song, & Li, 2011). The high ability of MAE in enhancing extraction yield is because of the molecular

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interactions between the electric component of the microwave field with the dipolar molecules and the ionic species present in the extraction mixtures (solvent-sample). This technology through penetrating energy into solid materials in terms of nonionizing radiation in a spectral

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frequency of 300-300,000 MHz generates volumetrically-distributed heating because of the molecular friction (Rodriguez-Jasso, Mussatto, Pastrana, Aguilar, & Teixeira, 2011). This mechanism originates from the dipolar rotation of polar solvents and the conductive migration of dissolved ions, enhancing the mass transfer coefficient of target ingredients (Pandit, Vijayanand, & Kulkarni, 2015). Thus, the direct effect of microwaves on molecules through the simultaneous

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occurrence of dipole polarization and ionic conduction can change microwave energy into thermal one, leading to an almost immediate heating up of the sample to extract bioactive compounds within material matrix toward the solution. MAE is thus considered as a promising technique for extracting polysaccharides with significant bioactivities (Bhatia, Sharma, Nagpal, & Bera, 2015; Wang et al., 2018; Zhao, Zhang, & Zhou, 2019). The microwave irradiation with the deep rupture of the cell wall at short extraction times improves the yield and bioactivity of polysaccharides as a result of the effect of microwaves on both the solvent (volumetric heating) 3

and the sample (improved liberate of polysaccharides from the matrix into the solvent) (Soria, Ruiz-Aceituno, Ramos, & Sanz, 2015). A number of structural factors such as monosaccharide composition, uronic acids content, molecular weight (Mw), glycosidic bond type in the backbone chain, and the esterification degree are profoundly affected on the antiradical, antioxidant, and antimicrobial activities of polysaccharides extracted from biological sources (Zhang, Lv, He, Shi, Pan, & Fan, 2013). The polysaccharides extracted by MAE show the excellent biological properties owing to the integrity of molecular structures in terms of functional glycosidic

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linkages with a higher Mw, and uronic acid content. Despite the critical role of MAE as a novel technology in the extraction of bioactive

polysaccharides, there is still no review study about the effects of microwave waves in

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improving the biological properties of these macromolecules. With the current review, the aim was to present a comprehensive overview of the progress so far made regarding the application

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of MAE and its principal operating parameters in improving biofunctional properties such as antioxidant, antiradical, and antibacterial, antifungal, and antiviral activities of polysaccharides

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from different sources.

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Power

Solvent

Temp

Time

Open Magnet Stirrer

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ON

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Sample

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Magnet stirrer

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2. In vitro antioxidant activity

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Fig. 1. A schematic representation of the MAE system for polysaccharides

2.1. Nitrite scavenging and phospho-molybdenum reduction activities In the food industry, nitrite is one of the most common ingredients in meat processing to

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intensify the color of final products and to extend their shelflife period. However, the procarcinogenic agent of nitrosamine can be generated via the chemical reaction between nitrite and amine constitutes present in protein-rich substances, medicines, and pesticide residues. Nitrosamine can be linked to diazoalkane, proteins, and intracellular components in the body and increased the cell damage and cancer risk (Lee, Kim, Jeong, & Park, 2006). Hence, it is

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interesting to find the ingredients with protective functions through scavenging nitrosamine or its precursor, reducing the formation of cancer cells (Gharibzahedi, Razavi, & Mousavi, 2013; Javanbakht et al., 2017). A potent nitrite scavenging activity (NSA) was reported for polysaccharides extracted from Fructus Meliae Toosendan (Xu, Yu, Wang, Xu, & Liu, 2018), and four seaweeds including Durvillaea antarctica, Gracilaria lemaneiformis, Sarcodia ceylonensis, and Ulva lactuca (He, Xu, Chen, & Sun, 2016) using MAE. A dose-dependent NSA behavior for polysaccharides extracted from F. Meliae Toosendan (1.2-2.0 mg/mL) and the

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different seaweeds (1.0-4.0 mg/mL) was observed. The maximum NSA value (67.3%) of F. Meliae Toosendan polysaccharide was found at the concentration of 2.0 mg/mL (Xu, Yu, et al., 2018). The lowest (< 25%, at 1.0 mg/mL) and highest (~52%, %, at 4.0 mg/mL) NSAs among seaweeds were for polysaccharides obtained from D. antarctica and S. ceylonensis, respectively (He et al., 2016). Nevertheless, He et al. (2016) reported that the effectiveness order of seaweed polysaccharides to induce NSA as follow: S. ceylonensis > U. lactuca > D. antarctica > G. lemaneiformis. Besides, the NSA ability of seaweeds was weaker than their potential to quench

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free radicals. The high presence of sulfated groups in the structure of polysaccharides, especially at more concentrations has an essential role in scavenging nitrite groups. Also, the hydroxyl

groups of polysaccharides can typically prevent the development of nitrosamines by donating

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hydrogens (Wang, Li, Zeng, & Liu, 2008; Zhang, Wang, Wang, Liu, Hou, & Zhang, 2010). On the other hand, Preethi and Mary Saral (2016) evaluated the phospho-molybdenum reduction

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activity (PMRA) of three polysaccharide fractions (PDP-1, PDP-2, and PDP-3) isolated from the fruits of Pithecellobium dulce. A concentration-dependent PMRA pattern was identified in the

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range of 2-10 mg/mL. They reported a high potential for the quantitative reduction of

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molybdenum (VI) to molybdenum (V) (~68-75%) for the three fractions at 10 mg/mL.

2.2. Inhibition of lipid peroxidation and erythrocyte hemolysis The inhibition activities of polysaccharides obtained from the fruit bodies of Auricularia

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auricular (Zeng et al., 2012) and fresh stems of Dendrobium devonianum (Li, Li, Peng, Xie, Ruan, & Huang, 2018) on lipid peroxidation were assessed based on the thiobarbituric acid method. Zeng et al. (2012) declared that the polysaccharide of A. auricular in a dose-dependent pattern could significantly increase the inhibition of lipid peroxidation (ILP) in egg yolk homogenate. The minimum (47.6%) and maximum (80.4%) values of ILP were recorded at

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concentrations of 0.0625 and 1.0 mg/mL, respectively. The potential of ILP in this study was more than that of polysaccharides obtained from A. auricular by Fan, Zhang, Yu, and Ma (2007). This discrepancy may be due to the difference in the extraction method (hot-water extraction (HWE) vs. MAE) and Mw of extracted polysaccharides. The lower Mw of polysaccharides obtained by MAE probably caused a significant increase in antioxidant activities because these biomolecules have a high number of free hydroxyl groups which can decrease the viscosity and increase the solubility of compounds (Zhang, Lv, Song, Jin, Huang, Fan, & Cai, 2015). The 6

inhibition of FeCl2-induced lipid peroxidation by the polysaccharides extracted from nonpuffed (44.5%) and puffed (47.6%) strips of D. devonianum (particularly in higher concentrations, 2.0 mg/mL) may be due to the high number of hydroxyl groups (Li et al., 2018). The erythrocyte membrane composes of polyunsaturated fatty acids susceptible to free radical-induced peroxidation. The presence of bioactive polysaccharides due to the high number of uronic acids containing both hydroxyl and carboxylic acid functional groups can quench free radicals, preventing the hemolysis of blood cells (Cheng et al., 2013). Fernandes, Filipe, Coelho, and

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Manso (1991) earlier mentioned that the ILP was accompanied by the inhibition of hemolysis in red-cells. For the first time, Cheng et al. (2013) evaluated the erythrocyte hemolysis inhibition (EHI) activity of polysaccharides (Epimedium acuminatum) extracted by MAE on blood samples

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collected from a healthy volunteer. The hemolysis was induced by 2,2'-azobis(2-

amidinopropane) dihydrochloride (AAPH), which is a known model to assess the oxidative

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damage of cells membrane. They pointed out that the polysaccharide fractions (EAP- (E, H, M, U)) from E. acuminatum had a protective effect through the hemolysis inhibition of human red

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blood cells. This antioxidant capacity decreases the formation of free radicals mediated by AAPH, which may be attributed to the reduction of oxidative injury of biological membranes

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(Ma, Liu, Zhou, Yang, & Liu, 2000).

2.3. Reducing power of ferric ions

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The ferric reducing antioxidant power (FRAP) of polysaccharides obtained from E. acuminatum (Cheng et al. 2013), paddlefish cartilage (Zhang, Zhao, Xiong, Huang, & Shen, 2013), tangerine peels (Chen, Jin, Tong, Lu, Tan, Tian, & Chang, 2016), bamboo leaves (Zhang, Li, Zhong, Peng, & Sun, 2016), and okra (Yuan et al., 2019) by MAE method was evaluated. A concentrationdependent manner was found for the reducing colorless Fe(III) to colored Fe(II)-

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tripyridyltriazine form according to the absorbance measured at 593 nm. An increase in concentration of polysaccharides of E. acuminatum (0-1.0 mg/mL), tangerine peels and bamboo leaves (0-5.0 mg/mL), and okra (0.5-4.5 mg/mL) could significantly improve the FRAP. The okra polysaccharide extracted by MAE had a higher FRAP compared to that obtained by HWE and PWE at all the used concentrations was found (Yuan et al., 2019). It seems that these electron-donating antioxidant components because of the presence of a high number of uronic acids in their structure are able to reduce ferric ions. Furthermore, the use of microwave radiation 7

reduces the MW of extracted polysaccharides through changing the structural configuration by loosening the matrix of the cell wall and separating the parenchyma cells and thus enhances the extraction yield and antioxidant activity (Kaufmann & Christen, 2002). In the reducing power (RP) assay, the presence of antioxidant polysaccharides reduces the Fe(III)/ferricyanide complex to the form of Fe(II). As a result, the level of Fe(II) in antioxidant solutions can be checked by determining the formation of Prussian blue at 700 nm. In recent years, numerous studies have been assessed the reducing power of polysaccharides extracted by

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MAE (Table 1). The hydrogen-donating ability through active hydroxyl and carboxyl groups present in the

molecular enables polysaccharides to reduce Fe(III)/ferricyanide ions. This antioxidant capacity

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also acts through breaking the free-radical chain and preventing the peroxide formation via the reaction with specific precursors of peroxide (Esfehani, Ghasemzadeh, & Mirzadeh, 2018;

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Gharibzahedi, 2017; Wang, Zhang, Wang, Zhao, Wu, & Yao, 2009; Wang, Zhang, Zhao, Wang, X., Wu, & Yao, 2010; Zhang, Lv, et al., 2013). Yuan and Macquarrie (2015), and He et al.

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(2016) showed that the sulfation could highly improve the RP of polysaccharides extracted from seaweeds of Ascophyllum nodosum, S. ceylonensis, G. lemaneiformis, D. antarctica, and U.

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lactuca. In general, there was a dose-dependent manner in RP of polysaccharides so that the highest used concentration showed the maximum RP. However, the difference in RP of different polysaccharides can be due to the discrepancy in their

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degree of substitutions, Mw, monosaccharide composition, and glycosidic linkages (Wang, Hu, Nie, Yu, & Xie, 2016). Among all the studied carbohydrate polymers, polysaccharides extracted from Gentiana scabra bge (Cheng, Zhang, Song, Zhou, Zhong, Hu, & Feng, 2016) at 1.2 mg/mL, Lilium davidii var. unicolor Salisb (Zhao, Zhang, Guo, & Wang, 2013) at 4.0 mg/mL, Chuanminshen violaceum (Dong, Zhang, et al., 2016) at 4.0 mg/mL, and A. auricular and Panax

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ginseng (Zeng et al., 2012; Zhao et al., 2019) at 10 mg/mL exhibited the most potent RP. Therefore, these biomacromolecules are high-potential photochemical ingredients to be formulated in potent therapeutic drugs and care products in modern medicine. Yuan and Macquarrie (2015) evaluated the MAE times (5, 15, and 30 min) and temperatures (90, 120, and 150°C) on the RP of fucoidan extracted from A. nodosum. They proved that the RP of fucoidan increases with a decrease in MAE temperature due to the more sulfate content (Hu, Liu, Chen, Wu, & Wang, 2010; Yang, Liu, Wu,

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Chen, & Wang, 2011). Besides, 15 min was considered an optimal time to extract fucoidan using MAE method (Yuan & Macquarrie, 2015).

Table 1. A summary of dose-dependent reducing power of polysaccharides extracted by MAE

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Polysaccharide source Concentration (mg/mL) a Maximum RP (Ǻ700 nm) Reference Artemisia sphaerocephala 0.1-5.0 0.86 Wang et al. (2009) Potentilla anserina 0.0-5.0 0.87 Wang et al. (2010) Poria cocos Wolf 0.0-2.0 ~0.68-0.72 Wang, Zhang, et al. (2016) Passiflora edulis Sims 0.25-1.5 0.32 Xiong, Li, Zheng, Hu, Cui, and Li (2019) Auricularia auricular 0.5-10.0 ~0.92-0.96 Zeng et al. (2012) Chuanminshen violaceum 0.06-4.0 0.306 Dong, Lin, et al. (2016) C. violaceum 0.06-4.0 ~1.0 Dong, Zhang, et al. (2016) Flammulina velutipes 0.31-5.0 0.126 Zhang, Lv, et al. (2013) Gentiana scabra bge 0.0-1.2 0.99 Cheng et al. (2016) Actinidia chinensis Planch. 1.0-3.0 0.71 Han et al. (2019) Glycyrrhiza uralensis 0.1-4.0 ~0.19-0.38 Wang et al. (2018) Sarcodia ceylonensis 1.2-4.0 ~0.18 He et al. (2016) Gracilaria lemaneiformis 1.2-4.0 ~0.12 He et al. (2016) Durvillaea antarctica 1.2-4.0 ~0.40 He et al. (2016) Ascophyllum nodosum 3.0 ~0.62-0.65 Yuan and Macquarrie (2015) Ulva prolifera 2.0 ~0.27 Yuan et al. (2018) Ulva lactuca 1.2-4.0 ~0.22 He et al. (2016) Ulva pertusa 0.5-3.0 ~0.95 Le, Golokhvast, Yang, and Sun (2019) Fructus Meliae Toosendan 0.2-1.0 ~0.152 Xu, Yu, et al. (2018) Ziziphus jujube 0.025-2.0 ~0.60-0.70 Rostami and Gharibzahedi (2016) Hippophae rhamnoides 0.05 ~0.18-0.28 Wei et al. (2019) Lilium davidii 0.1-3.0 ~1.2 Zhao et al. (2013) Panax ginseng 1.0-10.0 ~1.6 Zhao et al. (2019) Lentinus edodes 0.5-3.0 0.309 Yin, Fan, Fan, Shi, and Gao (2018) Grifola frondosa 0.4-2.0 0.477 Chen, Ji, et al. (2019) Agaricus blazei Murrill 0.2-2.0 ~0.85-0.90 Zhang, Lv, Pan, Shi, and Fan (2011) Berberis dasystachya Maxim 0.05-3.2 0.549 Han, Suo, Yang, Meng, and Hu (2016) Pithecellobium dulce 0.2-10.0 ~0.68-0.75 Preethi and Mary Saral (2016) a The best dose detected to show the reducing power was the upper limit of the selected concentration range.

2.4. Ferrous metal ions chelating activity

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A dose-dependent manner in the ferrous ion chelating activity (FICA) was found for polysaccharides extracted from Artemisia sphaerocephala (Wang et al., 2009), Potentilla anserina (Wang et al., 2010), himematsutake (Zhang, Lv, Pan, Shi, & Fan, 2011), Flammulina velutipes (Zhang, Lv, et al., 2013), L. davidii var. unicolor Salisb (Zhao et al., 2013), Poria cocos Wolf (Wang, Zhang, et al., 2016), Dendrobium officinale (He et al., 2018), bamboo shoots (Chen, Fang, et al., 2019), seabuckthorn (Wei et al., 2019), and Chlorella vulgaris (Yu et al., 2019) by MAE. It was demonstrated that the delay in the metal-catalyzed oxidation could be

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improved with increasing the polysaccharide concentration. Although the FICA of these biomacromolecules was desirable, this antioxidant index in most polysaccharides was determined to be lower than EDTA. Naughton and Grootveld (2001) explained that the FICA of EDTA is due to the presence of functional groups containing nitrogen and carboxyl. However, the FICA of polysaccharides may also be ascribed to the hydroxyl groups and the formation of cross-bridge between carboxyl groups in the structure of uronic acids and divalent ions of iron (Wang et al., 2009; Yu et al., 2019).

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Accordingly, Wei et al. (2019) mentioned that the increase of microwave power by 650 W could significantly reduce the FICA of polysaccharides extracted from seabuckthorn berries with an enhancement in the cleavage rate of glucosidic bonds and a decrease in the number of hydroxyl

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groups. Generally, the FICAs of A. sphaerocephala and Chimonobambusa quadrangularis polysaccharides obtained by MAE were higher than those of the same polysaccharides extracted

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by HWE (Chen, Fang, et al., 2019; Wang et al., 2009). This fact may be owing to the microwave heating, causing polarization of polar bonds (like the C-O-C glycosidic linkage) and the

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enhancement of molecular reactivity (Zhang, Lv, et al., 2013). The chelating ability of Fe(II) ions by chelators through forming σ bonds is an essential biological defense mechanism because this

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function can remove Fe(II) ions participating in hydroxyl radical-generating Fenton-type reactions. The cell protection against oxidative damages thus can be possible by minimizing Fe(II) ions and inhibiting the formation of reactive oxygen species (ROSs) and lipid peroxidation

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(Gharibzahedi, 2018; Zhang, Lv, et al., 2013).

3. In vitro antiradical activity

3.1. 1,1-diphenyl-2-picrylhydrazyl (DPPH·) radical scavenging activity DPPH· is a stable nitrogen-centered on free radical with an unpaired electron which is

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responsible for the strong absorbance at 517 nm. The number of this free radical can easily decrease in the exposure to proton radical scavengers. The deep-purple solution of DPPH· can be changed to yellow as it is quenched by antioxidants (Sharma & Bhat, 2009). DPPH· was the most common free radical used to determine antiradical and antioxidant activities of polysaccharides extracted by MAE (Table 2). The maximum DPPH· radical scavenging percentage of an infinite number of polysaccharides extracted under the optimum conditions accompanied by their monosaccharide composition and MW is given in Table 2. According to 10

the tested concentrations, the most potent polysaccharides extracted by MAE to quench DPPH· radical were obtained from E. acuminatum (~92%, at 0.004 mg/mL), okra pods (67.0%, at 0.018 mg/mL), Palmaria palmata (51.32%, at 0.025 mg/mL), Hippophae rhamnoides L. (55%, at 0.05 mg/mL), Sargassum thunbergii (90.8-95.2%, at 0.4 mg/mL), Opuntia ficus indica (82.0%, at 0.4 mg/mL), Zizyphus jujuba Mill. (67.0-74.1%, at 0.2 mg/mL), C. vulgaris (65.1%, at 0.4 mg/mL), and mung bean hulls (83.2%, at 0.8 mg/mL) (Table 2). As a result, these biomacromolecules could be employed as a potential antioxidative and nutraceutical ingredient in designing

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functional foods, medicinal products, and pharmaceutical formulations (Izadi, Khedmat, & Mojtahedi, 2019; Xiong, Li, Zheng, Hu, Cui, & Li, 2019). Similar to the other antioxidant assessments, a concentration-dependent behavior was observed to scavenge DPPH·. The

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presence of electrophilic groups (like keto or aldehyde) in acidic polysaccharides can improve the DPPH· scavenging ability through facilitating the hydrogen release from O–H bond. Hence,

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the enhanced hydrogen-donating ability at high polysaccharide concentrations is related to the high number of unmethylated uronic acids and hydroxyl groups (Song, Chen, Li, Jia, & Zhong,

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2018; Yin, Fan, Fan, Shi, & Gao, 2018). Maintaining the triple helical structure integrity in extracted polysaccharides with lower MWs also can significantly intensify their potential to

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scavenge DPPH·. Under this condition, smaller polysaccharide fractions due to their larger surface area have a greater chance to contact with the different types of radicals such as DPPH· (Chen, Fang, et al., 2019). Chen, Xu, and Zhu (2010) also explained that the low ratio of glucose

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in the glycosidic structure might represent better antioxidant ability.

3.2. Hydroxyl (OH·) radical scavenging activity Since the highly active OH· radical can quickly react with any biomolecules, it may cause severe damages to the adjacent tissues or organs. Hence, finding the biomaterials with removal ability

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of OH· radical is necessary for antioxidant defense in cell systems. Table 2 shows the highest OH· scavenging activity of polysaccharides extracted by MAE at the optimal concentration. At the high concentration of 10 mg/mL, the maximum OH· inhibition rate for polysaccharides extracted from A. sphaerocephala, P. anserina, Lycium ruthenicum, and A. auricular was 50.1, 50.1, ~68, and ~95%, respectively (Table 2). However, some polysaccharides extracted by MAE exhibited a considerable OH· scavenging activity at very low doses, including H. rhamnoides at 0.08 mg/kg (~90%), Carex meyeriana Kunth at 0.2 mg/kg (~75%), C. vulgaris at 0.4 mg/kg 11

(56.2%), and S. thunbergii at 0.8 mg/kg (68.7-72.4%) (Table 2). Two mechanisms involved in this antioxidant activity are the generation suppression of OH· and the cleaning of formed radicals (Qu, Yu, Jin, Wang, & Luo, 2013; Wang et al., 2009). In general, an increase in the concentration of polysaccharides extracted by MAE could significantly improve the inhibition rate of OH· radical. The addition of electron-donating substituent enhanced the inhibition rate of OH· owing to the increased electron density on the heterocyclic ring of the carbons (Jeong, Seo, & Jeong, 2009). Therefore, the more hydrogen donation to OH· radicals by crude and purified

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polysaccharides can produce stable radicals to block the chain reaction. This antiradical activity is highly associated with the iron-chelating ability of polysaccharides (Wang et al., 2010; Wei et al., 2019). This radical type plays a significant role in superoxidation by H2O2 with metal ions

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(e.g., ferrous or copper). Accordingly, polysaccharides with metal chelating capacity render them inactive in Fenton reaction may have a high ability to inhibit OH· radical (Zhang, Lv, et al.,

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2013). He et al. (2018) have recently evaluated the H2O2 scavenging potential of polysaccharides extracted from D. officinale stem by MAE. They found that the use of 5 mg/mL of this

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biopolymer could notably improve the H2O2 scavenging activity (> 80%). Instead, the low activity in scavenging OH· by some polysaccharides probably is because of the formation of

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strong intermolecular and intramolecular hydrogen bonds, leading to the reactivity inhibition of

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hydroxyl groups in the polymer chains (Cheng et al., 2016; Jeong et al., 2009).

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Table 2. The DPPH· and OH· radical scavenging activities of different polysaccharides extracted under the optimal MAE conditions

Okra pods

14.79

Passiflora edulis Sims Auricularia auricular Bamboo shoots

ND ND 9.94

Chuanminshen violaceum

34.59

S/M, 40:1; MP, 450 W; ETe, 65°C; ETi, 15 min

Flammulina velutipes Premna microphylla Turcz Mung bean hulls

12.67

S/M, 30:1; MP, 500 W; ETe, 110°C; ETi, 10 min S/M, 20:1; MP, 720 W; ETe, 60°C; ETi, 1.66 min

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Coreopsis Tinctoria

18.25 60.03d

S/M: 17:1; 700 W; ETi, 70 s

4.26

S/M: 59:1; 500 W; ETi, 6.5 min

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9.95

Maximum DPPH· scavenging activity (at an optimal dose) 52.32% (at 2.0 mg/mL)

ro

Poria cocos Wolf

MW (Da) 0.375149.14×105 0.1834.528×105

-p

13.33

Monosaccharide composition (molar ratio, or %) b L-Ara (1.0), D-Xyl (4.98), D-Lyx (1.69), L-Man (27.86), L-Glc (3.76), L-Gal (13.92) ND c

Maximum OH· scavenging activity (at an optimal dose) 50.1% (at 10.0 mg/mL)

Reference Wang et al. (2009)

66.01% (at 10.0 mg/mL)

50.1% (at 10.0 mg/mL)

Wang et al. (2010) Wang, Zhang, et al. (2016) Samavati (2013)

Man (4.02%), Glc (79.48%), Gal (4.93%), Ara (11.57%)

15.1×103

92.5% (at 2.0 mg/mL)

~100% (at 1.0 mg/mL)

ND

ND

67.0% (at 180 μg/mL)

ND

9.24×104

68.39% (at 1.5 mg/mL)

ND

2.77×104

75.4% (at 10.0 mg/mL)

~95% (at 10.0 mg/mL)

13.607×104

~68% (at 4.0 mg/mL)

~72% (at 4.0 mg/mL)

2.11×1034.06×105

41.40% (at 2.0 mg/mL)

ND

Dong, Lin, et al. (2016)

ND

64.34% (at 2.5 mg/mL)

33.40% (at 2.5 mg/mL)

18.35×103

90.0% (at 1.0 mg/mL)

ND

ND

83.2% (at 800 μg/mL)

80.31-85.48% (at 5.0 mg/mL)

4.336×105

~77% (at 1.2 mg/mL)

ND

Zhang, Lv, et al. (2013) Lu, Li, Jin, Li, Yi, and Huang (2019) Zhong, Lin, Wang, and Zhou (2012) Guo et al. (2019)

re

Potentilla anserina

Optimal MAE conditions a S/M, 32.8:1; MP, 323 W; ETe, 60°C; ETi, 70 min S/M, 14.5:1; MP, 369 W; ETe, 63.3°C; ETi, 76.8 min S/M, 20:1; MP, 800 W; ETe, MF, 2450 MHz; ETi, 2.0 min S/M, 5:1; MP, 395.56 W; ETe, 73.33°C; ETi, 67.11 min S/M, 27:1; MP, 420 W; ETi, 3.0 min MP, 860 W; ETe, 95°C; ETi, 25 min; pH, 7.0 S/M, 20:1; MP, 400 W; ETe, 90°C; ETi, 15 min

ND

lP

Yield (%) 31.81

Glc (37.53), Gal (1.0), Man (4.32), Ara (0.93), Rha (0.91) Man (1.85%), Rha (0.88%), Glc (33.81%), Gal (20.89%), Xyl (15.22%) Ara (20.48%), GlcA (1.17%), GalA (5.7%) Ara (2.25%), Xyl (4.26%), Man (7.41%), Gal (38.62%), Glc (47.15%) Fuc (18.49%), D-Man (20.54%), D-Glc (31.46%), D-Gal (29.35%) Rha (2.96), Ara (1.17), Man (1.04), Glc (8.07), Gal (2.05) + GalA (82.75%, pectin-type) Rha (1.0), Ara (0.12-0.3), Man (0.3-1.0), Gal (0.5)

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Polysaccharide source Artemisia sphaerocephala

Glc (1.0), GalA (2.04), Gal (2.41), Ara (1.94), Rha (0.19), Man (0.22), GlcA (0.09), Xyl (0.05)

Xiong et al. (2019) Zeng et al. (2012) Chen, Fang, et al. (2019)

13

Table 2. Continued

viper’s bugloss

25.11

Chuanminshen violaceum

34.59

S/M, 61.4:1; MP, 769.2 W; ETe, 42.3°C; ETi, 73.8 min S/M, 40:1; MP, 466 W; ETe, 64.5°C; ETi, 15 min

Carex meyeriana Kunth

0.28

S/M, 30:1; MP, 500 W; ETe, 95°C; ETi, 60 min

Glycyrrhiza uralensis Ascophyllum nodosum

4.23

S/M, 13:1; MP, 600 W; ETe, 70°C; ETi, 85 min ETe, 120°C; ETi, 15 min

Grifola frondosa Fructus Meliae Toosendan

36.38

S/M, 20:1; 0.01 M HCl, MF, 2.45 GHz; MP, 500 W; ETe, 120°C, ETi, 15 min

Jo

Ulva prolifera

16.08

7.36

15.75

Ara (2.56%), Xyl (2.813.78%), Man (4.19-6.25%), Gal (40.58-49.45%), Glc (43.55-46.83%) Rha (1.0), Xyl (1.95), Ara (1.72), Fru (1.78), Man (4.36), Glc (6.18) Ara (0.48-16.6%), Glc (48.8866.42%), Gal (19.12-23.97%) Fuc (8.12-42.25%), Rha (0.601.75%), Gal (5.01-12.23%), Glc (5.01-12.23%), Xyl 2.3718.72(%), Man (10.4123.19%), GlcA (22.21-62.0%) Man (0.04-1.05%), Rha (9.2942.17%), GlcA (2.58-22.77%), GalA (0.24-0.91%), Glc (24.37-87.22%), Gal (0.101.90%), Xyl (0.58-7.54%), Ara (0.16-0.93%) ND

S/M, 15:1; ETe, 75°C, ETi, 210 min S/M, 30:1; MP, 700 W; ETe, 60°C, ETi, 20 min

ND

of

S/M, 52.9:1; MP, 443.2 W; ETi, 7.6 min

3.8×104

Maximum DPPH· scavenging activity (at an optimal dose) 80.81% (at 1.2 mg/mL)

ro

2.92

MW (Da)

1.689×105

Maximum OH· scavenging activity (at an optimal dose) ~80% (at 1.2 mg/mL)

Reference Cheng et al. (2016)

~62-63% (at 3.0 mg/mL)

ND

Han et al. (2019)

ND

89.3% (at 5.6 mg/mL)

88.2% (at 4.0 mg/mL)

Tahmouzi (2014)

0.02311.02×103

42.44-67.91% (at 4.0 mg/mL)

ND

Dong, Zhang, et al. (2016)

15-913×103

~75% (at 0.2 mg/mL)

~75% (at 0.2 mg/mL)

4.51×103 2.08×105 M* 1.3437.54×103

68.23% (at 4.0 mg/mL)

~35% (at 4.0 mg/mL)

30.44% (at 10.0 mg/mL)

ND

Hu, Wang, Zhou, and Li (2018) Wang et al. (2018) Yuan and Macquarrie (2015)

10.5227×103

27.6% (at 2.0 mg/mL)

ND

Yuan et al. (2018)

ND

94.94% (at 2.0 mg/mL)

73.89% (at 2.0 mg/mL)

1.288×103

73.4% (at 1.0 mg/mL)

53.6% (at 2.0 mg/mL)

Chen, Ji, et al. (2019) Xu, Yu, et al. (2018)

-p

Kiwifruit

Monosaccharide composition (molar ratio, or %) b Man (1.0), Rha (9.89), GalA (51.59), Glc (35.37), Gal (38.06), Ara (99.13), Fuc (21.34) Man (0.36), Rha (0.23), GalA (3.28), Glc (1.0), Gal (1.69), Xyl (0.24), Ara (0.93) ND

re

Optimal MAE conditions a S/M, 34:1; MP, 157.09 W; ETi, 130.38 s

lP

Yield (%) 15.03

ur na

Polysaccharide source Gentiana scabra bge

14

Table 2. Continued

1993 mg/L*

S/M, 15:1; ETe, 70°C, ETi, 40 min

Cyphomandra betacea

36.52

S/M, 40:1; MP, 400 W; ETe, 60°C, ETi, 120 min

Hippophae rhamnoides Lilium davidii

0.264

Camptotheca acuminata Tangerine

8.61

Ulva pertusa

41.91

Sargassum thunbergii

2.84

S/M, 10:1; MP, 600 W; ETe, 85°C, ETi, 6 min S/M, 65:1; MP, 597 W; ETe, 50°C, ETi, 60 min S/M, 40:1; MP, 600 W; ETe, 70°C, ETi, 14 min S/M, 30:1; MP, 704 W; ETe, 52.2°C, ETi, 41.8 min S/M, 55.45:1; MP, 600 W; ETi, 43.63 min; pH, 6.57 S/M, 27:1; MP, 547 W; ETe, 80°C, ETi, 23 min

Lycium ruthenicum

8.25

36.55

Jo

19.9

3.9×103 11.95×104

S/M, 31.5:1; MP, 544 W; ETi, 25.8 min

ND

Maximum DPPH· scavenging activity (at an optimal dose) 51.32% (at 25 μg/mL)

Maximum OH· scavenging activity (at an optimal dose) 7.81% (at 93 μg/mL)

Li et al. (2011)

38.7, 43.9% (at 2.0 mg/mL)

74.4, 82.1% (at 2.0 mg/mL)

Lin et al. (2019)

67.0-74.1% (at 200 μg/mL)

ND

Rostami and Gharibzahedi (2016) Zhang et al. (2016)

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Man (8.22,10.46%), Rib (1.33,10.31%), GlcA (1.14%), Glc (71.26,72.50%), Gal (3.44,12.71%), Ara (0.52,4.54%), Fuc (3.60%) Glc (1.2-1.4), Ara (1.8-2.1), Gal (4.1-4.2), Rha (0.9-1.1)

9.1×104 1.5×105

Reference

mg/g: Man (6.9-14.6), Fuc (7.8-13.0), Xyl (40.9-109.0), Glc (167-246.7), Gal (167.0289.2), Ara (258.0-414.6) ND

ND

> 85% (at 10.0 mg/mL)

ND

ND

~60% (at 10.0 mg/mL)

ND

L-Rha (1.0), D-Man (6.89), DGlc (1.62), D-Gal (13.52) Glc (5.17): Man (4.82)

3.3×103 96.83×104 1.193×105

~55% (at 0.05 mg/mL)

~90% (at 0.05 mg/mL)

Kumar, Sivakumar and Ruckmani (2016) Wei et al. (2019)

~65% (at 3.0 mg/mL)

~72% (at 3.0 mg/mL)

Zhao et al. (2013)

Glc (1.0): Man (0.78)

121.34×103

82.22% (at 1.25 mg/mL)

74.96% (at 1.25 mg/mL)

GalA (42.5%), Ara (23.0%), Gal (20.0%), Rha (6.9%), Glc (4.2%), Man (3.5%) ND

17.8×103

82.5% (at 2.0 mg/mL)

95.7% (at 4.0 mg/mL)

Hu, Zhao, et al. (2019) Chen et al. (2016)

ND

46.51% (at 0.8 mg/mL)

ND

Le et al. (2019)

Ara (1.94-2.81%), Gal (23.230.7%), Glc (2.92-4.54%), Xyl (20.8-23.2%), Man (17.622.8%), GalA (8.11-9.74%), GlcA (13.9-17.7%) ND

190.4×103

90.80-95.23% (at 0.4 mg/mL)

68.7-72.4% (at 0.8 mg/mL)

Ren, Chen, Li, Fu, You, and Liu (2017)

ND

~88% (at 10.0 mg/mL)

~68% (at 10.0 mg/mL)

Liu et al. (2013)

ur na

Phyllostachys acuta

MW (Da)

ro

9.02

Monosaccharide composition (molar ratio, or %) b ND

-p

Zizyphus jujuba

2.12, 11.16

Optimal MAE conditions a S/M, 70:1; MP, 500 W; ETe, 70°C, ETi, 10 min EtOH conc., 26%; (NH4)2SO4 conc., 19.58%; ETe, 78.7°C, ETi, 19.55 min; S/M, 50:1 S/M, 30:1; MP, 400 W; ETe, 75°C, ETi, 60 min

re

Yield (%) 17.01

lP

Polysaccharide source Palmaria palmata Lentinus edodes

15

Table 2. Continued

12.31

ND

S/M, 10:1; MP, 400 W; ETi, 15 min

Birch (lignin)

26.49

Ficus carica

9.62

Chlorella vulgaris

17.1

S/M, 10:1; MP, 700 W; ETe, 101°C, ETi, 30 min S/M, 15:1; MP, 600 W; ETi, 3.5 min; pH, 1.4 S/M, 30:1; ETe, 50°C, ETi, 30 min;

F. velutipes

ND

2.95×105, 1.52×103

S/M, 20:1; MP, 650 W; ETe, 50°C, ETi, 2 min

Maximum OH· scavenging activity (at an optimal dose) 82.7% (at 8.0 mg/mL)

Reference Zhang et al. (2011)

62.37% (at 12.0 mg/mL)

ND

Han et al. (2016)

84.3% (at 2.0 mg/mL)

ND

Xu, Hou, Hu, and Liu (2018)

3.67×106 M*

82.0% (at 400 μg/mL)

ND

ND

7.2910.86×103M*

75-87% (at 4.0 mg/mL)

ND

GlcA (0.3), GalA (3.3), Glc (0.8), Fuc (0.5), Ara (0.3), Gal (1.0), Rha (0.2), Man (0.1) Ara (0.01), Gal (2.15), Man (1.59), Xyl (0.73), Glc (1.0), Rha (0.99), Fuc (0.06) Glc (4.24-24.17), Gal (2.0513.98), Man (1.82-3.67), Xyl (1.0)

6.25×103

~68% (at 15.0 mg/mL)

ND

Salehi, EmamDjomeh, Askari, and Fathi (2019) Zhou, Liu, Wang, Xu, and Sun (2012) Gharibzahedi et al. (2019a)

1.1-4.29×103

65.1% (at 0.4 mg/mL)

56.2% (at 0.4 mg/mL)

Yu et al. (2019)

29.9362.29×103

47.38-65.34% (at 8.0 mg/mL)

85.41-88.21% (at 8.0 mg/mL)

Liu, Zhang, Ibrahim, Gao, Yang, and Huang (2016) Preethi and Mary Saral (2016) Cheng et al. (2013)

ur na

Opuntia ficus indica

Rha (1.0), Ara (17.3), Xyl (1.33), Man (7.0), Glc (2.33), Lac (1.78) L-Rha (9%), L-Ara (32%), DGal (20%), D-Glc (55%), DXyl (19%), D-Man (2%), DGlcA (14%), D-GalA (1%) Glc (78.0%), Ara (12.9%), Xyl (4.8%), Gal (2.4%), Man (2.4%)

ND

Maximum DPPH· scavenging activity (at an optimal dose) ~50% (at 1.0 mg/mL)

of

Eucommia ulmoides Oliver

MW (Da)

ro

6.472

Monosaccharide composition (molar ratio, or %) b ND

-p

Berberis dasystachya Maxim

Optimal MAE conditions a S/M, 32.7:1; MP, 400 W; ETe, 74.64°C, ETi, 29.37 min S/M, 25.84:1; MP, 433.13 W; ETi, 35.18 min S/M, 29:1; ETe, 74.0°C, ETi, 15 min

12.055×103, 38.83×103 M*

re

Yield (%) 12.35

lP

Polysaccharide source Agaricus blazei Murrill

S/M, 4:1; MP, 350 Xyl (-), Man (-), Gal (-), Rha ND ~ 65-73% (at 10.0 μg/mL) ND W; ETe, 50°C, ETi, (-), Glc (-), Rib (-) 180 min Epimedium ND S/M, 10:1; ETi, 60 ND ND ~ 92% (at 4.0 μg/mL) ND acuminatum min a S/M: solvent: material (v/w), MP: microwave power (W), E Te: Extraction temperature (°C), ETi: Extraction time (min), MF: Microwave frequency (MHz) b

ND

Jo

Pithecellobium dulce

Ara: arabinose, Xyl: xylose, Lyx: lyxose, Man: mannose, Glc: glucose, Gal: galactose, Rha: rhamnose, GlcA: glucuronic acid, GalA: galacturonic acid, Fuc,

fucose, Fru: fructose, Rib: ribose, Lac: lactose; c ND: not detected; d Based on mg glucose equivalent (GE)/g dry weight (DW)

16

3.3. ABTS radical scavenging activity ABTS· (2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) is a nitrogen-centered synthetic radical cation which is produced through oxidizing ABTS with potassium persulphate. An antioxidant component can convert ABTS· to its non-radical form by donating an electron. This antioxidant assay can be measured according to the reduction of the blue-green solution containing ABTS· radical cation at 734 nm. In recent years, the polysaccharides extracted from natural sources by MAE demonstrated the

of

considerable ABTS radical scavenging activities (Fig. 2). Some of these algal and plant sources were A. auricular (IC50 = 1.23 mg/mL; Zeng et al., 2012), E. acuminatum (36.63% at 2.0

mg/mL; Chen et al., 2013), Berberis dasystachya Maxim (77.09% at 12 mg/mL; Han, Suo,

ro

Yang, Meng, & Hu, 2016), seaweeds of S. ceylonensis, U. lactuca L., and D. antarctica (IC50 = 3.59-3.99 mg/mL; He et al., 2016), C. violaceum (95.29% at 2.0 mg/mL; Dong, Lin, et al.,

-p

2016), C. violaceum (98.82-100% at 10.0 mg/mL; Dong, Zhang, et al., 2016), F. velutipes (97.03-98.92% at 6.0 mg/mL; Liu, Zhang, Ibrahim, Gao, Yang, & Huang, 2016), C. meyeriana

re

Kunth (IC50 = 0.0399 mg/kg; Hu et al., 2018), D. officinale (IC50 = 2.659 mg/mL; He et al., 2018), U. prolifera (68.6% at 2.0 mg/mL; Yuan et al. 2018), C. quadrangularis (IC50 = 0.582

lP

mg/mL; Chen, Fang, et al., 2019), common fig (~ 75% at 15 mg/mL; Gharibzahedi et al., 2019a), snow chrysanthemum (IC50 ≤ 1.121 mg/mL; Guo et al., 2019), kiwifruit (~72% at 3.0 mg/mL; Han et al., 2019), U. pertusa (50.37% at 0.8 mg/mL; Le et al., 2019), okra (IC50 = 2.50-4.40

ur na

mg/mL; Yuan et al., 2019), and P. ginseng (97.09% at 10.0 mg/mL; Zhao et al. 2019). Results showed that the polysaccharides extracted from U. prolifera and C. meyeriana Kunth had substantial scavenging capacity against ABTS radical (Fig. 2a and 2b). The radical scavenging of ABTS· by polysaccharides in most studies was lower than the positive control compounds (PCCs (e.g., vitamin C and BHA)). But it was reported that the

Jo

polysaccharide fractions extracted from C. violaceum, F. velutipes, and P. ginseng had an equal power or even more compared to vitamin C to scavenge ABTS· radical (Dong, Zhang, et al. 2016; Liu et al., 2016; Zhao et al., 2019). This result might be attributable to the ABTS· radical being more suitable for determining hydrophilic antioxidant compounds compared to the other free radicals like DPPH (Floegel, Kim, Chung, Koo, & Chun, 2011; Gharibzahedi et al., 2013). Optimizing the MAE conditions could prevent the production of polysaccharides with a high denaturation rate or weak structural integrity (Gharibzahedi, Smith, & Guo, 2019b). Yuan et al. 17

(2018) found that the high sulfur content in the structure of U. prolifera polysaccharides compared to the low MW had a more pronounced role in improving the ABTS· radical scavenging activity. The presence of keto or aldehyde groups, high content of uronic acids, and plentiful monosaccharide components may increase the inhibition activities of ABTS· radical by acidic polysaccharides (Hu et al., 2018; Song et al., 2018).

lP

5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0

Jo

ur na

b

ABTS scavenging activity (IC50, mg/mL)

re

-p

ro

of

a

Polysaccharide source

Fig. 2. The ABTS radical scavenging activity of polysaccharides extracted by MAE (a, percentage (%); b, IC50 value (mg/mL)) 3.4. Superoxide anion (O2·−) radical scavenging activity 18

The superoxide radical (O2·−) is one of the most important highly toxic radical species. This radical is produced by a high number of biological and photochemical reactions (Wang et al., 2018). It is considered as one of the precursors of the singlet oxygen and OH· radicals, and thus can indirectly cause the lipid peroxidation. In this antioxidant assay, 1,2,3-phentriol is rapidly autoxidized in an alkaline solution and developed intermediate products (e.g., O2·−). The presence of antioxidant components like polysaccharides through scavenging O2·− can interfere with 1,2,3-phentriol autoxidation. Accordingly, one of the standard procedures to assess the

of

antioxidant capacity of polysaccharides is the inhibition activity of the self-oxidation of 1,2,3phentriol (Liu et al., 2013).

The polysaccharides obtained by MAE from the various bio-sources had a favorable potential to

ro

quench O2·− radical (Fig. 3). The results were as follows: A. sphaerocephala (52.86% at 0.5 mg/mL), A. auricular (81.7% at 2.0 mg/mL), C. quadrangularis (~67% at 4.0 mg/mL), C.

-p

vulgaris (61.2% at 0.4 mg/mL), F. Meliae Toosendan (50.3% at 1.0 mg/mL), G. scabra bge (46.84% at 1.2 mg/mL), Lentinus edodes (55.9-66.5% at 1.0 mg/mL), L. edodes (27.04% at 3.0

re

mg/mL), L. ruthenicum (53.01% at 5.0 mg/mL), G. uralensis (64.25% at 4.0 mg/mL), and L. davidii var. unicolor Salisb (53.6 and ~85% at 1.0 and 4.0 mg/mL, respectively), P. anserina

lP

(~53% at 1.0 mg/mL) (Chen, Fang, et al., 2019; Cheng et al., 2016; Lin et al., 2019; Liu e al., 2013; Wang et al., 2009; Wang et al., 2010; Wang et al., 2018; Xu, Yu, et al., 2018; Yin et al., 2018, Yu et al., 2019; Zhao et al., 2013; Zeng et al., 2012). Compared to the HWE method, the

ur na

polysaccharides extracted by MAE also had better ability to scavenge O2·− radicals (Chen et al., 2019; Wang et al., 2009; Wang et al., 2010). However, Yu et al. (2019) reported a similar activity to scavenge O2·− radical for bamboo shoots polysaccharides extracted by HWE and MAE methods. On the other hand, the oxygen radical absorbance capacity (ORAC) of C. quadrangularis (Chen et al., 2019), and Armillaria luteo-virens (Chen, Shao, Tao, & Wen, 2015)

Jo

polysaccharides extracted by MAE were evaluated. Chen et al. (2019) found that the polysaccharide extracted from C. quadrangularis by MAE (131.0 μmol Trolox equiv./g) had significantly higher ORAC values than that of obtained by HWE (107.7 μmol Trolox equiv./g). However, Chen et al. (2015) reported a lower ORAC (94.0 μmol Trolox equiv./g) for A. luteovirens polysaccharides. The potential of polysaccharides to scavenge O2·− radicals is due to the presence of some electrophilic groups (like keto or aldehyde group) in their molecular structure, facilitating the 19

release of hydrogen from O-H bond to be stabilized O2·− (Cheng et al., 2016). An increase in the concentration of extracted polysaccharides could considerably increase the scavenging activity of O2·− radical. Therefore, these bioactive macromolecules, particularly at higher concentrations might be helpful to avoid the damage induced by O2·− radicals under pathological conditions

5.0 4.5 4.0

of

80

Polysaccharide conc. (mg/mL, ●)

100

3.5

60

3.0

ro

2.5

40

2.0 1.5

20

-p

Superoxide anion (O2·−) radical scavenging activity (%, □)

(Wang et al., 2018; Zhang, Yu, Liang, & Chen, 2015).

0.5 0.0

lP

re

0

1.0

Polysaccharide source

Fig. 3. The scavenging activity of polysaccharides extracted by MAE against superoxide anion

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radical

3.5. Nitric oxide (NO·) radical scavenging activity The use of NO· scavenging activity test has received less attention to evaluating the antiradical property of polysaccharides extracted by MAE. Yuan et al. (2019) and Guo et al. (2019) have

Jo

recently assessed the NO· scavenging activities of polysaccharides extracted from okra and snow chrysanthemum by MAE technique, respectively. A dose-dependent manner to describe this property was exhibited. An increase in the concentration of okra (0.5-4.5 mg/mL) and snow chrysanthemum (0.1-0.6 mg/mL) polysaccharides could significantly lead to an increase in the NO· scavenging activity. Therefore, the best activity to scavenge NO· by okra (~75%, IC50 = 0.90 mg/mL) and snow chrysanthemum (~85%, IC50 = 0.105 mg/mL) polysaccharides was obtained at 0.6 and 4.5 mg/mL, respectively. In general, it was found that the polysaccharides 20

extracted by MAE had a better NO· scavenging activity compared to those obtained by PWE, UAE, and HWE methods. Therefore, the bioactive polysaccharides extracted by MAE are outstanding candidates for formulating nutraceutical products and medical drugs to prevent the incidence of different disorders and diseases such as hypertension, stroke, arteriosclerosis, cancer, and cardiovascular risks (Mirbabaei Ghafghazi, Javadi, Mirzadeh, & Yaseri, 2008; Noroozian, Raeesi, Hashemi, Khedmat, & Vahabi, 2018). A direct correlation was found between the formation of NO· radicals and immune responses (e.g., macrophage activation)

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during antitumor and antimicrobial functions. The production of pro-inflammatory biomarkers like NO and cytokines is begun in the presence of bioactive polysaccharides activating

macrophages (Hu, Zhao, et al., 2019). In a dose-dependent manner, the polysaccharides of

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Camptotheca acuminata, Diaphragma juglandis fructus and D. devonianum extracted by MAE released NO· of RAW 264.7 macrophage cells through the activation of nitric oxide synthase

-p

(iNOS) (Liu et al., 2017). Hence, the administration of these polysaccharides at high

invasion by tumors and pathogens.

lP

4. In vivo antioxidant activity

re

concentrations might cause the speed-up the delivery of antigens to resist against the foreign

Recently, the in vivo antioxidant evaluations of polysaccharides extracted by MAE have conducted. The oxidative stress resistance in terms of methyl viologen induction (MVI),

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hydrogen peroxide induction (HPI), and antioxidant biomarkers including malonaldehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), and protein carbonyls (PCO) were determined (Han, Li, Ding, Xiong, & Zhao, 2017; Wei et al., 2019; Yu et al., 2019). In the MVI and HPI tests, Yu et al. (2019) initially treated small, transparent nematode worms (Caenorhabditis elegans) with M9 buffer, and solutions of metformin hydrochloride and C.

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vulgaris polysaccharide, incubated (at 20°C for 48 h), added oxidative stress agents, including MV (83 mg/mL) or H2O2 (0.1 M), and finally cultured at 20°C. The survivability rate of nematodes was determined during the 40 min time intervals. Results showed that the survival time (ST) of the nematodes against MV could prolong by exposing them with the six polysaccharide fractions. They mentioned that the three polysaccharides of AEPs, CEPs, and MAPs compared to the control could notably increase the ST of nematodes. Although the protective effect of these polysaccharides against MV was comparable to vitamin C, this activity 21

was weaker than the deltamine compound. Besides, the polysaccharide fraction of UWPs from C. vulgaris typically improved the ST of the nematodes against H2O2 than the control group (Yu et al., 2019). Feng et al. (2018) earlier proved that the use of Panax notoginseng polysaccharide could increase the stress resistance in the nematode C. elegans. The enhanced activities of SOD, CAT, and GSH peroxidase (GSH-Px) can be highly associated with the immune improving activity of polysaccharides, whereas a rise in peroxidative damage can be represented with the increased production of MDA (as the most common final metabolite

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in lipid peroxidation of omega-3 and omega-6 fatty acids) and PCO (Haeri-Araghi, Zarabadipour, Safarzadeh-Khosroshahi, & Mirzadeh, 2018; Wei et al., 2019; Yuan et al., 2009; Zhang, Li, Zhou, Lu, Xu, & Li, 2003). PCO is typically formed before the other biomarkers. It is

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an irreversible form of protein metabolism with more stability compared to MDA (Weber,

Davies, & Grune, 2015). Under a dose-dependent manner, an increase in the concentration of

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polysaccharides extracted from Hippophae rhamnoides L. led to a significant decrease in PCO level (Wei et al., 2019). Han et al. (2019) determined the MDA level, and SOD and GSH-Px

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activities in sera samples collected from male mice administered with rice bran polysaccharide (RBP) dissolved in physiological saline. They showed that the SOD and GSH-Px activities in the

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animals administered with RBP could be significantly improved with an increase in the polysaccharide dose. Also, a significant reduction in MDA level was observed at high concentrations of RBP. However, it was demonstrated that the polysaccharides extracted by EAE

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method at different doses (50-150 mg/kg) had a better potential to improve the SOD (206.46215.12 vs. 223.59-235.88 U/mL), and GSH-Px (103.29-131.45 vs. 127.39-139.60 U) activities, and to reduce the MDA (4.78-3.74 vs. 3.77-3.09 nmol/mL) concentration. This fact can be ascribed to lower MW of the polysaccharides extracted by EAE compared to MAE. The larger surface area of polysaccharides with lower MW possibly could highly scavenge oxygen radicals

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because of the improved activities of intracellular antioxidant enzymes (SOD and GSH-Px), and subsequently inhibited the lipid peroxidation based on the decreased MDA levels (Chen, Fang, et al., 2019; Fan et al., 2007). Wei et al. (2019) showed that the increase in the concentration (100400 mg/kg) of seabuckthorn berries polysaccharide could significantly reduce the MDA content. Yu et al. (2009) also reported that all the polysaccharides extracted from C. vulgaris improved CAT (21.18-26.76%) and SOD (6.73-7.96%) activities in the nematode C. elegans. Before, Jin and Ning (2012) realized that the polysaccharide extracted from seed cake of Camellia oleifera 22

Abel could promote the antioxidant defense system of C. elegans through increasing the SOD and CAT activities. A similar trend in increasing the SOD and GSH activities was found by increasing the oral dosage of purified polysaccharide of seabuckthorn berries, administered to the male mice (Wei et al., 2019).

5. Antimicrobial activity 5.1. Antibacterial effect

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The results of antibacterial studies showed that the natural polysaccharides had a high ability to inhibit the growth of a wide span of spoilage and infectious bacteria. Hu, Li, Chen, Zhang, and Zhang (2013) evaluated the in vitro antibacterial activities of exopolysaccharides and

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polysaccharides extracted from L. edodes and F. velutipes using the MAE at 480 W for 3 min with the solid/water ratio of 1:30 g/mL. Although the exopolysaccharide of F. velutipes had not

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any inhibitory effect on Escherichia coli and Staphylococcus aureus, the exopolysaccharide extracted from L. edodes at 4.8 mg/mL could inhibit E. coli and S. aureus. The minimum

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inhibitory concentration (MIC) of exopolysaccharides and polysaccharides obtained from L. edodes against E. coli and S. aureus were 0.15 and 2.4 mg/mL, and the same (4.8 mg/mL).

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Therefore, the exopolysaccharide extracted from L. edodes using MAE had the best antibacterial activity. Su, Liu, Li, and Zheng (2011) evaluated the in vitro antibacterial of Epimedium polysaccharides extracted under the optimal conditions (400 W, 10 min and the solid/water ratio

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of 1:50 g/mL). The MIC values of this polysaccharide against Bacillus subtilis, E. coli, and S. aureus were assessed to be 1.6, 0.8, and 3.2 mg/mL, respectively. Based on the disc diffusion method (inhibition zones in diameter, mm), Tahmouzi (2014) assessed the in vitro antilisterial activities of crude polysaccharides extracted from the flower of viper’s bugloss. Four species of Listeria including L. ivanovii, L. marthii, L. monocytogenes, and L. seeligeri) were selected.

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Among the different species, a potent inhibition was determined against L. ivanovii (10.76 mm) and L. monocytogenes (8.64 mm) using 5.0 mg/mL viper’s bugloss polysaccharide. An increasing dose-dependent antilisterial activity for this polysaccharide was obtained in the range of 1.0-5.0 mg/mL (Tahmouzi, 2014). Alboofetileh et al. (2019) have recently realized that the high antibacterial potential of fucoidans extracted from Nizamuddinia zanardinii at 2.0 mg/mL against the Gram-negative bacteria E. coli and Pseudomonas aeruginosa. They also reported that the fucoidans obtained by MAE due to the higher sulfate content compared to subcritical water 23

extraction (SWE) had more ability to inhibit E. coli. Zhang et al. (2017) evaluated the antibacterial activity of Cordyceps cicadae polysaccharide against a high number of pathogenic bacteria such as E. coli, S. aureus, S. epidermidis, B. subtilis, B. cereus, P. aeruginosa, Salmonella paratyphi, Streptococcus pyogenes, Enterococcus faecalis, and Klebsiella pneumonia. Among the investigated bacteria, E. coli, S. paratyphi and P. aeruginosa, S. aureus, and B. subtilis highly were inactivated by the extracted polysaccharide (inhibition zones > 20.0 mm). An interesting MIC value (0.10 mg/mL) was found for E. coli. Zhao et al. (2019) also

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determined the MIC values of the pathogenic strains of S. aureus, E. coli, B. subtilis, and B. pumilus in the exposure of P. ginseng polysaccharides extracted by MAE and HWE methods. They declared that the polysaccharide extracted by MAE had higher antibacterial ability than

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that of obtained by HWE. The MIC values of S. aureus, E. coli, B. pumilus, and B. subtilis were 0.25, 0.025, 0.01, and 0.5 mg/mL. Therefore, P. ginseng polysaccharides compared to C. cicadae

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ones even had better potential to inactive E. coli. The most important mechanisms involved in antibacterial activities of polysaccharides are the change of membrane permeability, the

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structural destruction of proteins and nucleic acids, the inhibition of enzymes function, the synthesis prevention of nucleic acids, and the damage enhancement of the cell wall

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(Balamayooran, Batra, Fessler, Happel, & Jeyaseelan, 2010). In general, preventing the growth of bacteria can be the increased cell permeability of C. cicadae polysaccharide via damaging the cell inner membranes. This fact caused cell death with an increase in structural lesions and the

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liberation of electrolytes, proteins, and other cell contents (Gharibzahedi & Mohammadnabi, 2016; Rahbarimanesh et al., 2019).

5.2. Antifungal effect

The in vitro antifungal ability of Epimedium polysaccharides by Su et al. (2011) exhibited that

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this biomacromolecule did not have any inhibitory effect on fungi species of Monilinia mali (Takahashi) Whetzel, Colletotrichum lindemuthianum, and Trichoderma. However, the Epimedium polysaccharide could well inhibit the growth of Aspergillus niger, Fusarium oxysporum sp. niveum, and Gibberella fujikuroi (Sawada) Wollenweber. Hu et al. (2013) also found that the exopolysaccharide obtained from L. edodes by MAE was able to inhibit Candida albicans with a MIC of 4.8 mg/mL. Also, Bhatia et al. (2015) using the disk diffusion assay explored that the in vitro antifungal activity of sulfated polysaccharides of porphyran derived 24

from the red algae Porphyra vietnamensis against C. albicans was much more than that of chitosan without any hemolytic toxicological effect. The antifungal potential was related to low MW and high content of sulfate groups of the extracted polysaccharides. Furthermore, the reduced/prevented hemolysis effect was attributed to the strong antioxidant activity of porphyran. Under the in vivo experiment, Bhatia et al. (2015) induced systemic candidiasis (5×104 spore/mL) in cyclophosphamide (CYP, 2.0 mg)-compromised mice, administration chitosan and porphyran, and then determined the sickness rate and survival responses. It was

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shown that the porphyran extracted by MAE had a stronger antifungal activity than chitosan with normalizing the excretion levels of urea, uric acid, and creatinine. The leading suppressing cause of oxidative stress injuries by porphyran probably is the inhibition of mitogen-activated protein

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kinase pathways through the deactivation of the extracellular kinase (Kim, Lee, & Lee, 2010).

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5.3. Antiviral effect

The antiviral activity assessment of polysaccharides is a significant concept to rational design top

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targeted medications. Alboofetileh et al. (2019) determined the strong in vitro antiviral activity of fucoidans extracted from N. zanardinii against the human herpes simplex virus 2 (HSV-2)

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infection (EC50 = 0.027-0.123 μg/mL). It was earlier demonstrated that the sulfated polysaccharides through linking to the molecules' anionic features could considerably prevent the virus adsorption (Mandal, Mateu, Chattopadhyay, Pujol, Damonte, & Ray, 2007). The

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antiherpetic (HSV-1) activities of carrageenans extracted from the red seaweed Solieria chordalis by MAE method were also reported by Boulho, Marty, Freile-Pelegrín, Robledo, Bourgougnon, and Bedoux (2017). The carrageenan fractions revealed a considerable EC50 (3.254.4 μg/mL) without any cytotoxicity effect. Increasing the antiviral activity at high extraction temperatures (105°C) and times (25 min) in MAE was because of the high degradation rate and

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low MW of S. chordalis carrageenan (Sun, Wang, Shi, & Ma, 2009). Moreover, they realized that the antiviral activity of carrageenan extracted by MAE was more than that of carrageenan obtained by the conventional technique (Boulho et al., 2017).

6. Conclusions and future directions The current overview highlighted the antioxidant, antiradical, and antibacterial, antifungal, and antiviral activities of polysaccharides extracted by MAE in both in vitro and in vivo studies. A 25

dose-dependent pattern was observed in most biofunctional evaluations so that an increase in the concentration of extracted polysaccharides could significantly improve the biological activities. The polysaccharides extracted by MAE had better bioactivity compared to those obtained by HWE. The various reasons of this processing superiority included: higher contents of sulfated and hydroxyl groups and unmethylated uronic acids, lower percentage of glucose in the monosaccharide composition, higher viscosity and solubility rate, better configuration changes facilitating the hydrogen liberation from O-H bonds, smaller molecular size or lower MW, and

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larger surface area to quench free radical species and to quickly diffuse into the cell membranes of microorganisms. Although the modeling tools such as response surface methodology (RSM) was used to optimize the operating parameters (e.g., microwave power, the solid/water ratio,

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extraction time, and extraction temperature) involved in MAE for reaching the highest extraction yield, very few studies have been conducted on the effect of MAE processing factors on the

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reviewed healthy-functional properties. According to existing studies and literature resources, serious attention should be given to the antimicrobial effect and NO· radical scavenging activity

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of polysaccharides extracted by MAE. Compared to the in vitro studies, less attention towards the evaluation of in vivo antioxidant and antimicrobial activities of polysaccharides extracted by

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MAE has been paid. It is thus necessary to assess the bioactivity of polysaccharides under the in vivo clinical conditions. The use of these functional polysaccharides in designing micro- and nano-capsules as bioactive cores or wall materials would be a promising approach in promoting

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public health with the controlled release in the gastrointestinal tract. Further investigations are needed to elucidate the relationship between chemical structure, biological activity, and

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processing effects of these biomacromolecules.

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References Alboofetileh, M., Rezaei, M., Tabarsa, M., Rittà, M., Donalisio, M., Mariatti, F., et al. (2019). Effect of different non-conventional extraction methods on the antibacterial and antiviral activity of fucoidans extracted from Nizamuddinia zanardinii. International Journal of Biological Macromolecules, 124, 131-137. Balamayooran, G., Batra, S., Fessler, M. B., Happel, K. I., & Jeyaseelan, S. (2010). Mechanisms of neutrophil accumulation in the lungs against bacteria. American Journal of Respiratory Cell

of

and Molecular Biology, 43(1), 5-16. Bhatia, S., Sharma, K., Nagpal, K., & Bera, T. (2015). Investigation of the factors influencing the molecular weight of porphyran and its associated antifungal activity. Bioactive

ro

Carbohydrates and Dietary Fibre, 5(2), 153-168.

Boulho, R., Marty, C., Freile-Pelegrín, Y., Robledo, D., Bourgougnon, N., & Bedoux, G. (2017).

-p

Antiherpetic (HSV-1) activity of carrageenans from the red seaweed Solieria chordalis

Applied Phycology, 29(5), 2219-2228.

re

(Rhodophyta, Gigartinales) extracted by microwave-assisted extraction (MAE). Journal of

Chen, C., Shao, Y., Tao, Y., & Wen, H. (2015). Optimization of dynamic microwave-assisted

lP

extraction of Armillaria polysaccharides using RSM, and their biological activity. LWT-Food Science and Technology, 64(2), 1263-1269.

Chen, G., Fang, C., Ran, C., Tan, Y., Yu, Q., & Kan, J. (2019). Comparison of different

ur na

extraction methods for polysaccharides from bamboo shoots (Chimonobambusa quadrangularis) processing by-products. International Journal of Biological Macromolecules, 130, 903-914. Chen, H., Xu, X. Q., & Zhu, Y. (2010). Optimization of hydroxyl radical scavenging activity of exo-polysaccharides from Inonotus obliquus in submerged fermentation using response surface methodology. Journal of Microbiology and Biotechnology, 20(4), 835-843.

Jo

Chen, R., Jin, C., Tong, Z., Lu, J., Tan, L., Tian, L., & Chang, Q. (2016). Optimization extraction, characterization and antioxidant activities of pectic polysaccharide from tangerine peels. Carbohydrate Polymers, 136, 187-197. Chen, X., Ji, H., Xu, X., & Liu, A. (2019). Optimization of polysaccharide extraction process from Grifola frondosa and its antioxidant and anti-tumor research. Journal of Food Measurement and Characterization, 13(1), 144-153.

27

Cheng, H., Feng, S., Jia, X., Li, Q., Zhou, Y., & Ding, C. (2013). Structural characterization and antioxidant activities of polysaccharides extracted from Epimedium acuminatum. Carbohydrate Polymers, 92(1), 63-68. Cheng, Z., Zhang, Y., Song, H., Zhou, H., Zhong, F., Hu, H., & Feng, Y. (2016). Extraction optimization, characterization and antioxidant activity of polysaccharide from Gentiana scabra bge. International Journal of Biological Macromolecules, 93, 369-380. Dong, H., Lin, S., Zhang, Q., Chen, H., Lan, W., Li, H., et al. (2016). Effect of extraction

of

methods on the properties and antioxidant activities of Chuanminshen violaceum polysaccharides. International Journal of Biological Macromolecules, 93, 179-185.

Dong, H., Zhang, Q., Li, Y., Li, L., Lan, W., He, J., et al. (2016). Extraction, characterization and

ro

antioxidant activities of polysaccharides of Chuanminshen violaceum. International Journal of Biological Macromolecules, 86, 224-232.

-p

Esfehani, M., Ghasemzadeh, S., & Mirzadeh, M. (2018). Comparison of fluoride ion concentration in black, green and white tea. International Journal of Ayurvedic Medicine, 9(4),

re

263-265.

Fan, L., Zhang, S., Yu, L., & Ma, L. (2007). Evaluation of antioxidant property and quality of

lP

breads containing Auricularia auricula polysaccharide flour. Food Chemistry, 101(3), 11581163.

Feng, S., Cheng, H., Xu, Z., Yuan, M., Huang, Y., Liao, J., et al. (2018). Panax notoginseng

ur na

polysaccharide increases stress resistance and extends lifespan in Caenorhabditis elegans. Journal of Functional Foods, 45, 15-23. Fernandes, A. C., Filipe, P. M., Coelho, H., & Manso, C. F. (1991). The inhibition of lipid peroxidation by cinnarizine possible implications to its therapeutic and side-effects. Biochemical Pharmacology, 41(5), 709-714.

Jo

Floegel, A., Kim, D. O., Chung, S. J., Koo, S. I., & Chun, O. K. (2011). Comparison of ABTS/DPPH assays to measure antioxidant capacity in popular antioxidant-rich US foods. Journal of Food Composition and Analysis, 24(7), 1043-1048. Gharibzahedi, S. M. T. (2017). Ultrasound-mediated nettle oil nanoemulsions stabilized by purified jujube polysaccharide: Process optimization, microbial evaluation and physicochemical storage stability. Journal of Molecular Liquids, 234, 240-248.

28

Gharibzahedi, S. M. T. (2018). The preparation, stability, functionality and food enrichment ability of cinnamon oil-loaded nanoemulsion-based delivery systems: A review. Nutrafoods, 17(2), 97-105. Gharibzahedi, S. M. T., & Mohammadnabi, S. (2016). Characterizing the novel surfactantstabilized nanoemulsions of stinging nettle essential oil: Thermal behaviour, storage stability, antimicrobial activity and bioaccessibility. Journal of Molecular Liquids, 224, 1332-1340. Gharibzahedi, S. M. T., & Mohammadnabi, S. (2017). Effect of novel bioactive edible coatings

of

based on jujube gum and nettle oil-loaded nanoemulsions on the shelf-life of Beluga sturgeon fillets. International Journal of Biological Macromolecules, 95, 769-777.

Gharibzahedi, S. M. T., Razavi, S. H., & Mousavi, S. M. (2013). Comparison of antioxidant and

ro

free radical scavenging activities of biocolorant synthesized by Dietzia natronolimnaea HS-1 cells grown in batch, fed-batch and continuous cultures. Industrial Crops and Products, 49, 10-

-p

16.

Gharibzahedi, S. M. T., Smith, B., & Guo, Y. (2019a). Pectin extraction from common fig skin

re

by different methods: The physicochemical, rheological, functional, and structural evaluations. International Journal of Biological Macromolecules, 136, 275-283.

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Gharibzahedi, S. M. T., Smith, B., & Guo, Y. (2019b). Ultrasound-microwave assisted extraction of pectin from fig (Ficus carica L.) skin: Optimization, characterization and bioactivity. Carbohydrate Polymers, 222, 114992.

ur na

Guo, H., Yuan, Q., Fu, Y., Liu, W., Su, Y. H., Liu, H., et al. (2019). Extraction optimization and effects of extraction methods on the chemical structures and antioxidant activities of polysaccharides from snow chrysanthemum (Coreopsis Tinctoria). Polymers, 11(2), 215. Haeri-Araghi, H., Zarabadipour, M., Safarzadeh-Khosroshahi, S., & Mirzadeh, M. (2018). Evaluating the relationship between dental caries number and salivary level of IgA in adults.

Jo

Journal of Clinical and Experimental Dentistry, 10(1), e66. Han, L., Suo, Y., Yang, Y., Meng, J., & Hu, N. (2016). Optimization, characterization, and biological activity of polysaccharides from Berberis dasystachya Maxim. International Journal of Biological Macromolecules, 85, 655-666. Han, Q. H., Liu, W., Li, H. Y., He, J. L., Guo, H., Lin, S., et al. (2019). Extraction optimization, physicochemical characteristics, and antioxidant activities of polysaccharides from kiwifruit (Actinidia chinensis Planch.). Molecules, 24(3), 461. 29

Han, W., Li, J., Ding, Y., Xiong, S., & Zhao, S. (2017). Structural features, antitumor and antioxidant activities of rice bran polysaccharides using different extraction methods. Journal of Food Science, 82(10), 2403-2410. He, J., Xu, Y., Chen, H., & Sun, P. (2016). Extraction, structural characterization, and potential antioxidant activity of the polysaccharides from four seaweeds. International Journal of Molecular Sciences, 17(12), 1988. He, L., Yan, X., Liang, J., Li, S., He, H., Xiong, Q., et al. (2018). Comparison of different

of

extraction methods for polysaccharides from Dendrobium officinale stem. Carbohydrate Polymers, 198, 101-108.

Hu, T. T., Liu, D., Chen, Y., Wu, J., & Wang, S. S. (2010). Antioxidant activity of sulfated

ro

polysaccharide fractions extracted from Undaria pinnitafida in vitro. International Journal of Biological Macromolecules, 46(2), 193-198.

-p

Hu, W., Zhao, Y., Yang, Y., Zhang, H., Ding, C., Hu, C., et al. (2019). Microwave-assisted extraction, physicochemical characterization and bioactivity of polysaccharides from

re

Camptotheca acuminata fruits. International Journal of Biological Macromolecules, 133, 127136.

lP

Hu, K., Liu, J., Li, B., Liu, L., Gharibzahedi, S. M. T., Su, Y., et al. (2019). Global research trends in food safety in agriculture and industry from 1991 to 2018: A data-driven analysis. Trends in Food Science & Technology,

ur na

Hu, Z., Wang, P., Zhou, H., & Li, Y. (2018). Extraction, characterization and in vitro antioxidant activity of polysaccharides from Carex meyeriana Kunth using different methods. International Journal of Biological Macromolecules, 120, 2155-2164. Hu, G. Y., Li, C. Y., Chen, M., Zhang, Q., & Zhang, Z. (2013). Extraction and antimicrobial activities of polysaccharide from Lentinus edodes and Flammulina velutipes. Journal of Wuhan

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Institute of Technology, 6.

Izadi, A., Khedmat, L., & Mojtahedi, S. Y. (2019). Nutritional and therapeutic perspectives of camel milk and its protein hydrolysates: A review on versatile biofunctional properties. Journal of Functional Foods, 60, 103441. Javanbakht, M., Akhavanmoghadam, J., Talaei, A. J., Aghyani, M., Mozafari, M., Khedmat, L., & Mohebbi, M. (2017). Differential expression of two genes Oct‐4 and MUC 5 AC associates

30

with poor outcome in patients with gastric cancer. Clinical and Experimental Pharmacology and Physiology, 44(11), 1099-1105. Jeong, J. B., Seo, E. W., & Jeong, H. J. (2009). Effect of extracts from pine needle against oxidative DNA damage and apoptosis induced by hydroxyl radical via antioxidant activity. Food and Chemical Toxicology, 47(8), 2135-2141. Jin, X., & Ning, Y. (2012). Antioxidant and antitumor activities of the polysaccharide from seed cake of Camellia oleifera Abel. International Journal of Biological Macromolecules, 51(4), 364-

of

368. Jin, X., & Ning, Y. (2012). Antioxidant and antitumor activities of the polysaccharide from seed cake of Camellia oleifera Abel. International Journal of Biological Macromolecules, 51(4), 364-

ro

368.

Kaufmann, B., & Christen, P. (2002). Recent extraction techniques for natural products:

-p

microwave‐assisted extraction and pressurised solvent extraction. Phytochemical Analysis: An International Journal of Plant Chemical and Biochemical Techniques, 13(2), 105-113.

re

Kim, K. J., Lee, O. H., & Lee, B. Y. (2010). Fucoidan, a sulfated polysaccharide, inhibits adipogenesis through the mitogen-activated protein kinase pathway in 3T3-L1

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preadipocytes. Life Sciences, 86(21-22), 791-797.

Kumar, S., Sivakumar, M., & Ruckmani, K. (2016). Microwave-assisted extraction of polysaccharides from Cyphomandra betacea and its biological activities. International Journal

ur na

of Biological Macromolecules, 92, 682-693.

Le, B., Golokhvast, K. S., Yang, S. H., & Sun, S. (2019). Optimization of microwave-assisted extraction of polysaccharides from Ulva pertusa and evaluation of their antioxidant activity. Antioxidants, 8(5), 129.

Lee, S. C., Kim, S. Y., Jeong, S. M., & Park, J. H. (2006). Effect of far-infrared irradiation on

Jo

catechins and nitrite scavenging activity of green tea. Journal of Agricultural and Food Chemistry, 54(2), 399-403. Li, T., Li, Y., Peng, J., Xie, Q., Ruan, R., & Huang, X. (2018). Microwave puffing assisted extraction of polysaccharides from Dendrobium devonianum. Journal of Food Processing and Preservation, 42(2), e13490. Li, Z. R., Wang, B., Zhang, Q. H., Huang, F. F., & Ma, J. H. (2011). Microwave-assisted extraction and the antioxidant activity of water-soluble polysaccharide from Palmaria palmata: 31

Extraction process and antioxidant activity of polysaccharide from Palmaria palmate. In 2011 5th International Conference on Bioinformatics and Biomedical Engineering (pp. 1-5). IEEE. Lin, Y., Zeng, H., Wang, K., Lin, H., Li, P., Huang, Y., et al. (2019). Microwave-assisted aqueous two-phase extraction of diverse polysaccharides from Lentinus edodes: Process optimization, structure characterization and antioxidant activity. International Journal of Biological Macromolecules, 136, 305-315. Liu, Q. M., Xu, S. S., Li, L., Pan, T. M., Shi, C. L., Liu, H., et al. (2017). In vitro and in vivo

of

immunomodulatory activity of sulfated polysaccharide from Porphyra haitanensis. Carbohydrate Polymers, 165, 189-196.

Liu, Y., Zhang, B., Ibrahim, S. A., Gao, S. S., Yang, H., & Huang, W. (2016). Purification,

ro

characterization and antioxidant activity of polysaccharides from Flammulina velutipes residue. Carbohydrate Polymers, 145, 71-77.

-p

Liu, Z., Dang, J., Wang, Q., Yu, M., Jiang, L., Mei, L., et al. (2013). Optimization of polysaccharides from Lycium ruthenicum fruit using RSM and its anti-oxidant activity.

re

International Journal of Biological Macromolecules, 61, 127-134.

Lu, J., Li, J., Jin, R., Li, S., Yi, J., & Huang, J. (2019). Extraction and characterization of pectin

131, 323-328.

lP

from Premna microphylla Turcz leaves. International Journal of Biological Macromolecules,

Ma, L., Liu, Z., Zhou, B., Yang, L., & Liu, Z. (2000). Inhibition of free radical induced oxidative

2056.

ur na

hemolysis of red blood cells by green tea polyphenols. Chinese Science Bulletin, 45(22), 2052-

Mandal, P., Mateu, C. G., Chattopadhyay, K., Pujol, C. A., Damonte, E. B., & Ray, B. (2007). Structural features and antiviral activity of sulphated fucans from the brown seaweed Cystoseira indica. Antiviral Chemistry and Chemotherapy, 18(3), 153-162.

Jo

Mirbabaei Ghafghazi, F., Javadi, M. A., Mirzadeh, M., & Yaseri, M. (2008). Ocular hypertension after myopic photorefractive keratectomy. Bina Journal of Ophthalmology, 13(2), 196-202.

Naughton, D. P., & Grootveld, M. (2001). EDTA bis-(ethyl phenylalaninate): a novel transition metal-ion chelating hydroxyl radical scavenger with a potential anti-inflammatory role. Bioorganic & Medicinal Chemistry Letters, 11(19), 2573-2575.

32

Noroozian, M., Raeesi, S., Hashemi, R., Khedmat, L., & Vahabi, Z. (2018). Pain: the neglect issue in old people’s life. Open Access Macedonian Journal of Medical Sciences, 6(9), 17731778. Pandit, S. G., Vijayanand, P., & Kulkarni, S. G. (2015). Pectic principles of mango peel from mango processing waste as influenced by microwave energy. LWT-Food Science and Technology, 64(2), 1010-1014. Preethi, S., & Mary Saral, M. (2016). Screening of natural polysaccharides extracted from the

of

fruits of Pithecellobium dulce as a pharmaceutical adjuvant. International Journal of Biological Macromolecules, 92, 347-356.

Qu, C., Yu, S., Jin, H., Wang, J., & Luo, L. (2013). The pretreatment effects on the antioxidant

ro

activity of jujube polysaccharides. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 114, 339-343.

-p

Rahbarimanesh, A., Mojtahedi, S. Y., Sadeghi, P., Ghodsi, M., Kianfar, S., Khedmat, L., Siyahkali, S. J. M., Yazdi, M. K., & Izadi, A. (2019). Antimicrobial stewardship program (ASP):

re

an effective implementing technique for the therapy efficiency of meropenem and vancomycin antibiotics in Iranian pediatric patients. Annals of Clinical Microbiology and Antimicrobials,

lP

18(1), 6.

Ren, B., Chen, C., Li, C., Fu, X., You, L., & Liu, R. H. (2017). Optimization of microwaveassisted extraction of Sargassum thunbergii polysaccharides and its antioxidant and

ur na

hypoglycemic activities. Carbohydrate Polymers, 173, 192-201. Rodriguez-Jasso, R. M., Mussatto, S. I., Pastrana, L., Aguilar, C. N., & Teixeira, J. A. (2011). Microwave-assisted extraction of sulfated polysaccharides (fucoidan) from brown seaweed. Carbohydrate Polymers, 86(3), 1137-1144. Rostami, H., & Gharibzahedi, S. M. T. (2016). Microwave-assisted extraction of jujube

Jo

polysaccharide: Optimization, purification and functional characterization. Carbohydrate Polymers, 143, 100-107. Rostami, H., & Gharibzahedi, S. M. T. (2017a). Cellulase-assisted extraction of polysaccharides from Malva sylvestris: Process optimization and potential functionalities. International Journal of Biological Macromolecules, 101, 196-206.

33

Rostami, H., & Gharibzahedi, S. M. T. (2017b). Mathematical modeling of mucilage extraction kinetic from the waste hydrolysates of fruiting bodies of Zizyphus jujuba mill. Journal of Food Processing and Preservation, 41(4), e13064. Salehi, E., Emam-Djomeh, Z., Askari, G., & Fathi, M. (2019). Opuntia ficus indica fruit gum: Extraction, characterization, antioxidant activity and functional properties. Carbohydrate Polymers, 206, 565-572. Samavati, V. (2013). Central composite rotatable design for investigation of microwave-assisted

of

extraction of okra pod hydrocolloid. International Journal of Biological Macromolecules, 61, 142-149.

Sharma, O. P., & Bhat, T. K. (2009). DPPH antioxidant assay revisited. Food Chemistry, 113(4),

ro

1202-1205.

Song, J., Chen, X., Li, R., Jia, Y., & Zhong, Q. (2018). Optimization microwave-assisted

American Journal of Pharmacy, 37(2), 401-407.

-p

extraction and antioxidant activity of polysaccharides from Gentiana macrophylla. Latin

re

Soria, A. C., Ruiz-Aceituno, L., Ramos, L., & Sanz, L. M. (2015). Microwave-assisted extraction of polysaccharides. Polysaccharides: Bioactivity and Biotechnology, 987-1008.

lP

Su, Y., Liu, P. J., Li, L. M., & Zheng, X. C. (2011). Extraction of Epimedium polysaccharides by microwave-assisted technology and the antibacterial activity. Food Science and Technology, 36(11), 158-161.

ur na

Sun, L., Wang, C., Shi, Q., & Ma, C. (2009). Preparation of different molecular weight polysaccharides from Porphyridium cruentum and their antioxidant activities. International Journal of Biological Macromolecules, 45(1), 42-47. Tahmouzi, S. (2014). Extraction, antioxidant and antilisterial activities of polysaccharides from the flower of viper's bugloss. International Journal of Biological Macromolecules, 69, 523-531.

Jo

Wang, B. S., Li, B. S., Zeng, Q. X., & Liu, H. X. (2008). Antioxidant and free radical scavenging activities of pigments extracted from molasses alcohol wastewater. Food Chemistry, 107(3), 1198-1204.

Wang, J., Zhang, J., Wang, X., Zhao, B., Wu, Y., & Yao, J. (2009). A comparison study on microwave-assisted extraction of Artemisia sphaerocephala polysaccharides with conventional method: Molecule structure and antioxidant activities evaluation. International Journal of Biological Macromolecules, 45(5), 483-492. 34

Wang, J., Zhang, J., Zhao, B., Wang, X., Wu, Y., & Yao, J. (2010). A comparison study on microwave-assisted extraction of Potentilla anserina L. polysaccharides with conventional method: Molecule weight and antioxidant activities evaluation. Carbohydrate Polymers, 80(1), 84-93. Wang, N., Zhang, Y., Wang, X., Huang, X., Fei, Y., Yu, Y., & Shou, D. (2016). Antioxidant property of water-soluble polysaccharides from Poria cocos Wolf using different extraction methods. International Journal of Biological Macromolecules, 83, 103-110.

of

Wang, J., Hu, S., Nie, S., Yu, Q., & Xie, M. (2016). Reviews on mechanisms of in vitro antioxidant activity of polysaccharides. Oxidative Medicine and Cellular Longevity, 2016, 5692852.

ro

Wang, Y., Li, Y., Ma, X., Ren, H., Fan, W., Leng, F., et al. (2018). Extraction, purification, and bioactivities analyses of polysaccharides from Glycyrrhiza uralensis. Industrial Crops and

-p

Products, 122, 596-608.

Weber, D., Davies, M. J., & Grune, T. (2015). Determination of protein carbonyls in plasma, cell

conditions. Redox Biology, 5, 367-380.

re

extracts, tissue homogenates, isolated proteins: focus on sample preparation and derivatization

lP

Wei, E., Yang, R., Zhao, H., Wang, P., Zhao, S., Zhai, W., et al. (2019). Microwave-assisted extraction releases the antioxidant polysaccharides from seabuckthorn (Hippophae rhamnoides L.) berries. International Journal of Biological Macromolecules, 123, 280-290.

ur na

Xia, E. Q., Wang, B. W., Xu, X. R., Zhu, L., Song, Y., & Li, H. B. (2011). Microwave-assisted extraction of oleanolic acid and ursolic acid from Ligustrum lucidum Ait. International Journal of Molecular Sciences, 12(8), 5319-5329. Xiong, F., Li, X., Zheng, L., Hu, N., Cui, M., & Li, H. (2019). Characterization and antioxidant activities of polysaccharides from Passiflora edulis Sims peel under different degradation

Jo

methods. Carbohydrate Polymers, 218, 46-52. Xu, J., Hou, H., Hu, J., & Liu, B. (2018). Optimized microwave extraction, characterization and antioxidant capacity of biological polysaccharides from Eucommia ulmoides Oliver leaf. Scientific Reports, 8(1), 6561. Xu, L., Yu, J. Q., Wang, X. Y., Xu, N., & Liu, J. L. (2018). Microwave extraction optimization using the response surface methodology of Fructus Meliae Toosendan polysaccharides and its antioxidant activity. International Journal of Biological Macromolecules, 118, 1501-1510. 35

Yang, Y. L., Liu, D., Wu, J., Chen, Y., & Wang, S. S. (2011). In vitro antioxidant activities of sulfated polysaccharide fractions extracted from Corallina officinalis. International Journal of Biological Macromolecules, 49(5), 1031-1037. Yin, C., Fan, X., Fan, Z., Shi, D., & Gao, H. (2018). Optimization of enzymes-microwaveultrasound assisted extraction of Lentinus edodes polysaccharides and determination of its antioxidant activity. International Journal of Biological Macromolecules, 111, 446-454. Yu, M., Chen, M., Gui, J., Huang, S., Liu, Y., Shentu, H., et al. (2019). Preparation of Chlorella

of

vulgaris polysaccharides and their antioxidant activity in vitro and in vivo. International Journal of Biological Macromolecules, 137, 139-150.

Yuan, C., Huang, X., Cheng, L., Bu, Y., Liu, G., Yi, F., et al. (2009). Evaluation of antioxidant

ro

and immune activity of Phellinus ribis glucan in mice. Food Chemistry, 115(2), 581-584.

Yuan, Q., Lin, S., Fu, Y., Nie, X. R., Liu, W., Su, Y., et al. (2019). Effects of extraction methods

-p

on the physicochemical characteristics and biological activities of polysaccharides from okra (Abelmoschus esculentus). International Journal of Biological Macromolecules, 127, 178-186.

re

Yuan, Y., & Macquarrie, D. (2015). Microwave assisted extraction of sulfated polysaccharides (fucoidan) from Ascophyllum nodosum and its antioxidant activity. Carbohydrate Polymers, 129,

lP

101-107.

Yuan, Y., Xu, X., Jing, C., Zou, P., Zhang, C., & Li, Y. (2018). Microwave assisted hydrothermal extraction of polysaccharides from Ulva prolifera: Functional properties and

ur na

bioactivities. Carbohydrate Polymers, 181, 902-910. Zeng, W. C., Zhang, Z., Gao, H., Jia, L. R., & Chen, W. Y. (2012). Characterization of antioxidant polysaccharides from Auricularia auricular using microwave-assisted extraction. Carbohydrate Polymers, 89(2), 694-700. Zhang, C. H., Yu, Y., Liang, Y. Z., & Chen, X. Q. (2015). Purification, partial characterization

Jo

and antioxidant activity of polysaccharides from Glycyrrhiza uralensis. International Journal of Biological Macromolecules, 79, 681-686. Zhang, L., Zhao, S., Xiong, S., Huang, Q., & Shen, S. (2013). Chemical structure and antioxidant activity of the biomacromolecules from paddlefish cartilage. International Journal of Biological Macromolecules, 54, 65-70.

36

Zhang, X., Li, M., Zhong, L., Peng, X., & Sun, R. (2016). Microwave-assisted extraction of polysaccharides from bamboo (Phyllostachys acuta) leaves and their antioxidant activity. BioResources, 11(2), 5100-5112. Zhang, Q., Li, N., Zhou, G., Lu, X., Xu, Z., & Li, Z. (2003). In vivo antioxidant activity of polysaccharide fraction from Porphyra haitanesis (Rhodephyta) in aging mice. Pharmacological Research, 48(2), 151-155. Zhang, Y., Wu, Y. T., Zheng, W., Han, X. X., Jiang, Y. H., Hu, P. L., et al. (2017). The

of

antibacterial activity and antibacterial mechanism of a polysaccharide from Cordyceps cicadae. Journal of Functional Foods, 38, 273-279.

Zhang, Z. F., Lv, G. Y., Song, T. T., Jin, Q. L., Huang, J. B., Fan, L. F., & Cai, W. M. (2015).

ro

Comparison of the preliminary characterizations and antioxidant properties of polysaccharides obtained from Phellinus baumii growth on different culture substrates. Carbohydrate Polymers,

-p

132, 397-399.

Zhang, Z., Lv, G., He, W., Shi, L., Pan, H., & Fan, L. (2013). Effects of extraction methods on

re

the antioxidant activities of polysaccharides obtained from Flammulina velutipes. Carbohydrate Polymers, 98(2), 1524-1531.

lP

Zhang, Z., Lv, G., Pan, H., Shi, L., & Fan, L. (2011). Optimization of the microwave-assisted extraction process for polysaccharides in himematsutake (Agaricus blazei Murrill) and

470.

ur na

evaluation of their antioxidant activities. Food Science and Technology Research, 17(6), 461-

Zhang, Z., Wang, F., Wang, X., Liu, X., Hou, Y., & Zhang, Q. (2010). Extraction of the polysaccharides from five algae and their potential antioxidant activity in vitro. Carbohydrate Polymers, 82(1), 118-121.

Zhao, B., Zhang, J., Guo, X., & Wang, J. (2013). Microwave-assisted extraction, chemical

Jo

characterization of polysaccharides from Lilium davidii var. unicolor Salisb and its antioxidant activities evaluation. Food Hydrocolloids, 31(2), 346-356. Zhao, J. L., Zhang, M., & Zhou, H. L. (2019). Microwave-assisted extraction, purification, partial characterization, and bioactivity of polysaccharides from Panax ginseng. Molecules, 24(8), 1605.

37

Zhong, K., Lin, W., Wang, Q., & Zhou, S. (2012). Extraction and radicals scavenging activity of polysaccharides with microwave extraction from mung bean hulls. International Journal of Biological Macromolecules, 51(4), 612-617. Zhou, S., Liu, L., Wang, B., Xu, F., & Sun, R. (2012). Microwave-enhanced extraction of lignin from birch in formic acid: Structural characterization and antioxidant activity study. Process

Jo

ur na

lP

re

-p

ro

of

Biochemistry, 47(12), 1799-1806.

38