Welcome to Chemistry and Industry of Forest Products,

Chemistry and Industry of Forest Products ›› 2020, Vol. 40 ›› Issue (3): 31-38.doi: 10.3969/j.issn.0253-2417.2020.03.004

Previous Articles     Next Articles

Analysis of Monosaccharide in Sesbania cannabina Seed Enzymatic Hydrolysate by Pre-column Derivatization Reversed-phase High Performance Liquid Chromatography

Qiannan ZHANG,Fangmin XU,Zhina LIAN,Xin LI*(),Qiang YONG   

  1. College of Chemical Engineering, Nanjing Forestry University, Nanjing 210037, China
  • Received:2019-12-13 Online:2020-06-28 Published:2020-06-29
  • Contact: Xin LI E-mail:xli@njfu.edu.cn

Abstract:

To establish a pre-column derivatization reversed-phase high performance liquid chromatography for the determination of mannose, glucose, galactose and xylose in the enzymatic hydrolysate of Sesbania cannabina seed, the pre-column derivatization was carried out by using 1-phenyl-3-methyl-5-pyrazolone (PMP) as derivatization reagent, and the effects of different chromatographic conditions were investigated. The experimental results showed that when the mobile phase was acetonitrile/potassium dihydrogen phosphate buffer solution (0.02 mol/L, pH value 6.5) and the derivatization conditions were PMP dosage 150 μL, reaction time 80 min, reaction temperature 60℃ and chloroform extraction for 3 times, the peak area of each monosaccharide component was larger and the separation effect was better. The methodological study verified that the R2 values of standard curves of four monosaccharide were over 0.999 4, the recovery rates were 96.9%-104.8%, the RSD values were lower than 0.56%, and the results repeatability, stability and precision tests were good, which indicated that the established method under these conditions was effective and convincible. The mass ratio of mannose, glucose, galactose and xylose was 1:0.44:1.92:0.53. In summary, pre-column derivatization high performance liquid chromatography provided a quantitative method with simple operation, accuracy, high resolution and good reproducibility for analyzing monosaccharide composition and content of the enzymatic hydrolysate of S. cannabina seed.

Key words: Sesbania cannabina seed, galactomannan, manno-oligosaccharides, hydrolysis, monosaccharide

CLC Number: