Abstract
The increasing interest of curdlan oligosaccharides (COS) in medicine and plant protection fields implies a necessity to identify and quantify this product. In the present study, an efficient and sensitive analytical method based on high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was established for the simultaneous separation and determination of D-glucose and ß-1,3-linked COS ranging from (COS)2 to (COS)6 within 20 min. Detection limits were 0.01 to 0.03 mg/L. The optimized assay was performed on a CarboPac-PA100 analytical column (4 mm × 250 mm) using isocratic elution with water−0.2 M sodium hydroxide−0.5 M sodium acetate mixture (50 : 30 : 20, v/v/v) as the mobile phase at a flow rate of 0.8 mL/min. Regression equations indicated a good linear relationship (R2 = 0.9992–1.0000, n = 6) within the test ranges. Quality parameters including precision and accuracy were fully validated and found to be satisfactory. More important, the regression of natural logarithm values of retention times (log10 RT) versus the degree polymerization (DP), as well as the slope coefficient of each COS's linear equation versus the corresponding DP, fitted a linear relationship well. These inherent linear relationships could provide valuable information to tentatively identify and quantify the COS even with the DP more than 6 without authentic standard. Furthermore, when the log10 RT was plotted against log10 flow rate for each COS, a perfect linear relationship was also observed.
Highlights
Curdlan, which is industrially manufactured as an exopolysaccharide of bacterium, is the only linear β-(1,3)-glucan homopolysaccharide without branching (Figure 1) [1,2,3]. is biopolymer is soluble in alkaline solutions and dimethylsulphoxide [4], but insoluble in water and most organic solvents due to the unique gel properties [5], which limits its wide biological utility and industrial applications
When applied for carbohydrates, High-performance anion exchange chromatography (HPAEC)-pulsed amperometric detection (PAD) affords highresolution separation and sensitive detection, which has been extensively used in carbohydrates analysis
Satisfactory chromatographic separation mainly depends on the column, mobile phase composition, and flow rate. ese variables were optimized using mixed D-glucose and (COS)2–6 standard solutions
Summary
Curdlan, which is industrially manufactured as an exopolysaccharide of bacterium, is the only linear β-(1,3)-glucan homopolysaccharide without branching (Figure 1) [1,2,3]. is biopolymer is soluble in alkaline solutions and dimethylsulphoxide [4], but insoluble in water and most organic solvents due to the unique gel properties [5], which limits its wide biological utility and industrial applications. Curdlan, which is industrially manufactured as an exopolysaccharide of bacterium, is the only linear β-(1,3)-glucan homopolysaccharide without branching (Figure 1) [1,2,3]. Curdlan has traditionally been used as a stabilizer, texturizer, and thickener in the food industry [6]. Ese hydrolyzed oligomers of curdlan exhibit remarkable biomedical functions, such as antitumor and immunological activities [18, 19]. COS can play an efficient role in plant protection and improvement for sustainable agriculture since it can efficiently activate the plant innate immune defense system prior to the infection of pathogens [20]. COS can elicit the plant’s natural defense responses in tobacco (Nicotiana tabacum L.) [21], potato McCain G1) [22], and Arabidopsis thaliana Col0 [23] against tobacco mosaic virus, Phytophthora infestans, and Botrytis cinerea infections, Journal of Analytical Methods in Chemistry
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