Separation and Determination of Monosaccharides with High Performance Liquid Chromatography by Using Internal Standard Method
高效液相色谱内标法分离和测定植物中的单糖

A new approach by which free plant monosaccharides were separated directly and determined with high performance liquid chromatography by using internal standard method was established and beta-indolylacetic acid was selected as the internal standard substance. The separation time for xylose(Xyl), fructose(Fru), glucose(Glu), sucrose(Suc), maltose(Mal), lactose(Lac) and raffinose(Raf) was within 18 min; and the detection limits were 1.8 micrograms, 2.3 micrograms, 2.7 micrograms, 1.8 micrograms, 3.5 micrograms, 4.1 micrograms and 4.3 micrograms respectively while the linear dynamics varied within the range of 5.0 micrograms/L-750 micrograms/L. In this work, the effects of the concentration of CH3CN and pH values in the mobile phase to the separation of the 7 sugars and beta-indolylacetic acid were studied as well. The method can be used to the determinations of jujube, apple samples. Moreover, the recovery experiment was also conducted. The results showed that the relative standard deviations of 5 determinations to Xyl, Fru, Glu and Suc were 1.66%-3.45%, 1.54%-3.01%, 1.29%-2.91%, 1.09%-2.65% and the recoveries were 97.4%-102.1%, 97.3%-101.8%, 98.7%-102.2%, 97.7%-102.5%, respectively.