According to the National Standards of China GB/T 23528.3-2025* “Quality Specifications for Oligosaccharides - Part 3: Xylooligosaccharides”, xylooligosaccharides content is determined using two high-performance liquid chromatography (HPLC) methods. In Appendix A, the content of each sugar component is calculated based on peak areas, using xylose as the reference standard. In Appendix B, xylooligosaccharides are decomposed into monosaccharides by acid hydrolysis. Then, xylose is quantitatively determined by HPLC.

*The version at the time of the application acquisition.

A cation-exchange gel column (Na‑form sulfonated styrene divinylbenzene copolymer; 8 mm (I.D.) x 300 mm (length); particle size: 6 µm; exclusion limit molecular weight: 10,000) or a column with equivalent performance is used. In Appendix C, chromatograms of actual samples analyzed using the SUGAR KS-802 column are presented. Several xylooligosaccharide standards were analyzed and SUGAR SC1211 confirmed the requirement was met.

System suitability requirement Difference between two replicate measurements under repeatability conditions: ≤ 5% of the mean of the two measurements

Sample: 20 µL each
Standard solutions

1. 1.Xylohexaose 0.2%
2. 2.Xylopentaose 0.2%
3. 3.Xylotetraose 0.2%
4. 4.Xylotriose 0.2%
5. 5.Xylobiose 0.2%
6. 6.Glucose 0.1%
7. 7.Xylose 0.1%
8. 8.L-Arabinose 0.1%

A cation-exchange gel column (H‑form sulfonated styrene divinylbenzene copolymer; 8 mm (I.D.) x 300 mm (length); particle size: 9 µm; exclusion limit molecular weight: 1,000) or a column with equivalent performance is used. Standard solution was analyzed using SUGAR SH1011. Xylose, L-arabinose, and glucose were well separated from each other, and it confirmed the requirements were met.

System suitability requirements Difference between two replicate measurements under repeatability conditions: ≤ 5% of the mean of the two measurements
Correlation coefficient for the calibration curve (xylose): ≥ 0.9990

Sample: 20 µL
Standard solution

1. 1.Glucose 0.2 mg/mL
2. 2.Xylose 0.5 mg/mL
3. 3.L-Arabinose 0.1 mg/mL

 

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