A method for the simultaneous determination of the eight monosaccharide components of Lycopus lucidus polysaccharides, utilizing pre-column derivatization of 1-phenyl-3-methyl-5-pyrazolone (PMP) reagent and high performance liquid chromatography (HPLC), is reported. The sample was degreased by petroleum ether and extracted by treatment with ultrasound waves, proteins were removed by treatment with the Sevage reagent, and a polysaccharide solution was prepared by precipitation with anhydrous ethanol. The polysaccharide solution was then treated with 2 mol/L trifluoroacetic acid (TFA) for 5 h at 110 ℃ to obtain the corresponding monosaccharide solution by hydrolysis. Finally, 0.5 mol/L of a methanolic solution of PMP was added to the monosaccharide solution kept in a water bath maintained at 70 ℃ for 1 h. All eight monosaccharides exhibited a linear relationship (r≥0.999 5) within their respective mass concentration ranges; the average recovery rate was 82.58%-108.71%, and the detection limit was 0.16-0.85 μg/mL. The polysaccharide extracted from Lycopus lucidus was mainly composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, arabinose, and phycosaccharide. Each of these monosaccharide components was present in an average ratio of 1.00:0.60:0.54:5.15:266.53:890.11:4.12:1.05. The average content of galactose and glucose, the major monosaccharide components in Lycopus lucidus polysaccharides, was 120.446 and 35.306 mg/g, respectively. Principal component analysis revealed that the monosaccharide content in Lycopus lucidus polysaccharides showed a trend of northern>southern>southwestern with respect to the different provinces in China. The method reported in this study is simple, sensitive, reproducible, practical, and reliable, and the derivatized substances can exhibit a high degree of stability. Therefore, this method is suitable for the identification and analysis of monosaccharide components in Lycopus lucidus polysaccharides and represents a reference standard for quality control of Lycopus lucidus polysaccharides used for medicinal purposes.
HUANG Xiaolan
,
HE Xufeng
,
YANG Qin
,
ZHANG Chunyi
,
ZHOU Nong
. Determination of monosaccharide composition in Lycopus lucidus polysaccharides by pre-column PMP derivatization and HPLC[J]. Food and Fermentation Industries, 2020
, 46(7)
: 250
-256
.
DOI: 10.13995/j.cnki.11-1802/ts.022499
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