Applications of asymmetrical flow field-flow fractionation for separation and characterization of polysaccharides: A review
Introduction
Polysaccharides are carbohydrates with complex molecular structures that are widely found in plants, animals, microorganisms, and other organisms. They not only act as a source of energy, but also participate in various activities of cells in life phenomena. Due to the special properties of polysaccharides and a variety of biological activities (such as rheological properties, adhesion, antioxidation, antitumor activity, immune regulation, and antiobesity), great interest has arisen in their properties and they have been widely used in food [1,2], medicine [3], [4], [5], materials [6,7], cosmetics [8,9] and other fields [10], [11], [12].
Polysaccharides are sugar chains composed of glycosidic bonds connecting monosaccharide units. The common glycosidic bonds are α-1,4-, β-1,4- and α-1,6-glycosidic bonds, and monosaccharides can be linked into straight chains or branched chains [13,14]. Polysaccharides usually have a complex structure and most of them have ultrahigh molecular weight (MW) in their primary structure. Related studies have shown that the conformation of polysaccharides often affects their natural biological activity, thus affecting their practical applications [15,16]. Therefore, accurate characterization of polysaccharide structures is particularly important. Size exclusion chromatography (SEC) is a technique for size-based separation in complex samples. It is also used in the separation of polysaccharides. However, the range of molar mass (generally > 106 Da) of SEC is limited, and the column adsorption effect and shear effect of the stationary phase limit its application in polysaccharides [17,18]. In recent years, asymmetrical flow field-flow fractionation (AF4) has attracted increasing attention due to the wide dynamic range (approximately from 1 nm up to 1 μm in the normal mode), and the use of ‘open channels’ without stationary phases or packing materials, which creates a wide range of selectivity in the analytical carrier liquid [19,20]. These characteristics of AF4 enable the sample to maintain not only good integrity but also good biological activity in the detection process, which is especially suitable for the analysis of complex analytes with fully preserved natural properties [20]. These advantages make AF4 widely use in the analysis and detection of biological macromolecules such as polysaccharides, proteins, and nucleic acids [21], [22], [23]. Moreover, the shear force is minimized in the AF4 channel compared to the SEC columns during separation which allows for a decrease in the impact of shear scission particularly on branched macromolecules and large macromolecules. In addition to separation, the size characterization of analytes by direct measurement of retention time is one of the key features of AF4. As a gentle separation technique, AF4 coupled with multiple detectors has been widely used in the separation and characterization of polysaccharides [24], [25], [26], [27].
In this review, the separation principle and elution mode of AF4 are briefly introduced, and an introduction to AF4 theory can be found elsewhere [21,27,28]. Early works on the characterization of polysaccharides by AF4 have been reviewed [18,29]. The focus of this review is on the development of AF4 in the separation and characterization of polysaccharides from different sources over the last decade to provide a basis for further research, production, and application of polysaccharides.
Section snippets
Principle of AF4
AF4 was first introduced by Wahlund and Giddings [19]. In this review, AF4 theory is described briefly. AF4 is a gentle and fast size-based separation method. As shown in Fig. 1(a), the upper wall of the AF4 channel is an impermeable polycarbonate glass plate, and the bottom channel plate is permeable and made of porous stainless steel frit material, a polyester trapezoidal spacer, and ultrafiltration membrane in the middle. The bottom channel plate and the ultrafiltration membrane form an
Optimization of operation conditions in AF4
In the process of AF4 separation, the resolution of the sample, sample recovery, and sample information obtained are affected by several factors, such as the carrier liquid, type of membrane, flow rate, channel thickness and type of detector. Therefore, it is necessary to optimize the operating conditions of AF4 to achieve good resolution and accurate sample information. The optimization of AF4 operation conditions depends on the surface properties of the sample. This section mainly reviews the
Polysaccharides from seed
Seed starch is one of the most abundant plant polysaccharides and has been widely studied as the most important source of human food energy. It is mainly composed of amylose and amylopectin. Amylose consists of linear chains of (1→4)-α-D-glucose linked residues with a small number of long chain branches and amylopectin also contains sequences of (1→4)-α-D-glucose linked units; however, it has extensive branching via (1→6)-α-D-glucose linkages. The literature [27,32] reviewed the application of
Summary and Outlook
AF4 has been widely used in the field of polysaccharides in recent years due to its wide separation range in size. AF4 coupled with multiple detectors (UV, MALS, dRI, ICP-MS, FL, QELS) provides more information on the sample such as Rh, Rg, MW, ρ, polymer conformation and branching degree of polysaccharides. For glycoproteins, AF4-FL can also be used for semiquantitative determination of polysaccharides. Compared to SEC, AF4 is more suitable for the separation of branched polysaccharides due to
Credit Author Statement
Xue Chen and Wenhui Zhang: writing-original draft. Yuwei Dou: Table data. Tiange Song: Drawing figure. Shigang Shen: Revising the manuscript. Haiyang Dou: Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgement
The authors acknowledge the support provided by the Nature Science Foundation of Hebei Province (B2016201002), the Key Project of Hebei Education Department (ZD2019009), the Medical Science Foundation of Hebei University (2020A08), and the Post-graduate's Innovation Fund Project (hbu2020ss056).
References (83)
- et al.
An overview of classifications, properties of food polysaccharides and their links to applications in improving food textures
Trends Food Sci. Technol.
(2020) - et al.
Characterization of molecular properties of wheat starch from three different types of breads using asymmetric flow field-flow fractionation (AF4)
Food Chem.
(2019) - et al.
Biological activities and pharmaceutical applications of polysaccharide from natural resources: A review
Carbohydr. Polym.
(2018) - et al.
Advanced nanomedicine characterization by DLS and AF4-UV-MALS: Application to a HIV nanovaccine, J
Pharm. Biomed. Anal.
(2020) - et al.
Ulvan, a bioactive marine sulphated polysaccharide as a key constituent of hybrid biomaterials: A review
Carbohydr. Polym.
(2019) - et al.
Asymmetrical flow field-flow fractionation coupled to inductively coupled plasma mass spectrometry for sizing SeNPs for packaging applications
Spectrochimica Acta Part B: Atomic Spectroscopy
(2017) - et al.
Prosopis juliflora as a new cosmetic ingredient: Development and clinical evaluation of a bioactive moisturizing and anti-aging innovative solid core
Carbohydr. Polym.
(2020) - et al.
Influence of extreme alkaline pH induced unfolding and aggregation on PSE-like chicken protein edible film formation
Food Chem.
(2020) - et al.
Bioactive polysaccharides from natural sources: A review on the antitumor and immunomodulating activities
Biocatal. Agric. Biotechnol.
(2019) Separation and characterization of food macromolecules using field-flow;fractionation: A review
Food Hydrocoll.
(2013)
Antitumor polysaccharides from mushrooms: a review on the structural characteristics, antitumor mechanisms and immunomodulating activities
Carbohydr. Res.
Characterisation of chitosan molecular weight distribution by multi-detection asymmetric flow-field flow fractionation (AF4) and SEC
Int. J. Biol. Macromol.
Flow field-flow fractionation: Recent applications for lipidomic and proteomic analysis, TrAC
Trends Anal. Chem.
Probing and quantifying DNA–protein interactions with asymmetrical flow field-flow fractionation
J. Chromatogr. A
Physicochemical and structural properties of starch from five Andean crops grown in Bolivia
Int. J. Biol. Macromol.
Co-elution phenomena in polymer mixtures studied by asymmetric flow field-flow fractionation
J. Chromatogr. A
Study on structure-function of starch by asymmetrical flow field-flow fractionation coupled with multiple detectors: A review
Carbohydr. Polym.
Field-flow fractionation: A gentle separation and characterization technique in biomedicine, TrAC
Trends Anal. Chem.
Fast molecular mass and size characterization of polysaccharides using asymmetrical flow field-flow fractionation-multiangle light scattering
J. Chromatogr. A
Rational strategy for characterization of nanoscale particles by asymmetric-flow field flow fractionation: A tutorial
Anal. Chim. Acta
Flow field-flow fractionation: Critical overview
J. Chromatogr. A
Investigation of steric transition with field programming in frit inlet asymmetrical flow field-flow fractionation
J. Chromatogr. A
Capillary electrophoresis and asymmetric flow field-flow fractionation for size-based separation of engineered metallic nanoparticles: A critical comparative review, TrAC
Trends Anal. Chem.
Study on steric transition in asymmetrical flow field-flow fractionation and application to characterization of high-energy material
J. Chromatogr. A
Hydrodynamic radius determination with asymmetrical flow field-flow fractionation using decaying cross-flows. Part II. Experimental evaluation
J. Chromatogr. A
Factors affecting measurement of channel thickness in asymmetrical flow field-flow fractionation
J. Chromatogr. A
Macromolecular structure and film properties of enzymatically-engineered high molar mass dextrans
Carbohydr. Polym.
Characterization of cereal β-glucan extracts: Conformation and structural aspects
Food Hydrocolloids
Analysis of β-glucan molar mass from barley malt and brewer's spent grain with asymmetric flow field-flow fractionation (AF4) and their association to proteins
Carbohydr. Polym.
Interaction between cereal β-glucan and proteins in solution and at interfaces
Colloids Surf. B. Biointerfaces
Flow optimisations with increased channel thickness in asymmetrical flow field-flow fractionation
J. Chromatogr. A
Colloidal features of softwood galactoglucomannans-rich extract
Carbohydr. Polym.
Characterization of a water soluble, hyperbranched arabinogalactan from yacon (Smallanthus sonchifolius) roots
Food Chem.
Molecular characterization of starches by AF4-MALS-RI: An alternative procedure
J. Cereal Sci.
Challenges in analysis of high-molar mass dextrans: Comparison of HPSEC, AsFlFFF and DOSY NMR spectroscopy
Carbohydr. Polym.
Molecular parameters of low methoxylated pectin affected by gelation with copper and cadmium cations
Bioact. Carbohydr. Diet. Fibre
Flow field-flow fractionation for the analysis and characterization of natural colloids and manufactured nanoparticles in environmental systems: A critical review
J. Chromatogr. A
Study on antidiabetic activity of wheat and barley starch using asymmetrical flow field-flow fractionation coupled with multiangle light scattering
J. Chromatogr. A
Characterization of the macromolecular and sensory profile of non-alcoholic beers produced with various methods
Food Res. Int.
Characterisation of cationic potato starch by asymmetrical flow field-flow fractionation. Influence of ionic strength and degree of substitution
Carbohydr. Polym.
Thermal, conformational and rheological properties of κ-carrageenan-sodium stearoyl lactylate gels and solutions
Carbohydr. Polym.
Cited by (18)
Ganoderma lucidum polysaccharide peptides GL-PPSQ<inf>2</inf> alleviate intestinal ischemia-reperfusion injury via inhibiting cytotoxic neutrophil extracellular traps
2023, International Journal of Biological MacromoleculesAsymmetrical flow field-flow fractionation combined with ultrafiltration: A novel and high-efficiency approach for separation, purification, and characterization of Ganoderma lucidum polysaccharides
2023, TalantaCitation Excerpt :Consequently, the small components are closer to the center of the channel due to their high diffusion coefficient and are eluted earlier than large components. AF4 as a gentle separation technique has been widely used for the purification of virus [26], sRNA [27], and the separation of polysaccharides [28]. In addition to separation, AF4 coupled with MALS and dRI detectors (AF4-MALS-dRI) can provide the information about the structure of polysaccharides (i.e., Mw and radius of gyration, Rg) [10,29].
Asymmetrical flow field-flow fractionation combined with liquid chromatography enables rapid, quantitative, and structurally informative detection of resistant starch
2022, Microchemical JournalCitation Excerpt :Asymmetrical flow field-flow fractionation (AF4) coupled online with multiangle light scatting (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI) has attracted increasing interest in the structure characterization of RS owing to its broad determination range in size [10]. However, the content of RS determined by AF4 is underestimation due to technical limitation in terms of utilization of ultrafiltration membrane as the accumulation wall [11–13]. The starch hydrolysis products with the size smaller than the molecular weight cutoff (MWCO) of ultrafiltration membrane can pass through AF4 accumulation wall, which results in an underestimation in determination of content of RS by AF4-dRI.
Study on effects of preparation method on the structure and antioxidant activity of protein-Tremella fuciformis polysaccharide complexes by asymmetrical flow field-flow fractionation
2022, Food ChemistryCitation Excerpt :In our previous study, LDL was enzymatically modified with phospholipase A2 (PLA2). The results showed that the stability of LDL modified PLA2 was enhanced (Chen et al., 2021a, b). In addition to enzyme modification, the addition of salt and sugar can also improve the thermal stability of LDL (Dixon & Cotterill, 1981; Xu et al., 2020).
Field flow fractionation (FFF): practical and experimental aspects
2022, Particle Separation Techniques: Fundamentals, Instrumentation, and Selected ApplicationsDifferent molecular sizes and chain conformations of water-soluble yeast β-glucan fractions and their interactions with receptor Dectin-1
2021, Carbohydrate PolymersCitation Excerpt :For WYG-3, ~1.9% of aggregates was detected by AF4-MALLS-RI, in contrast to no aggregates detected by SEC-MALLS-Vis-RI, indicating that AF4-MALLS-RI could detect aggregates more sensitively. There may be strong interactions between the hydroxyl groups of the polymer and the stationary phase packing of the column in SEC, but no stationary phase was used in AF4, so the influences of deformation and adsorption can all be reduced in AF4 experiment (Chen et al., 2021; Erber et al., 2009). For WYG-4 and WYG-5, the results of main part ➀ measured by AF4-MALLS-RI were basically consistent with those measured by SEC-MALLS-Vis-RI.