Yuanheng Guo and Bing Zhao* Pages 317 - 324 ( 8 )
Objective: A comprehensive monosaccharide quantifying method was developed in this article. Monosaccharides (including glucose, arabinose, galacturonic acid, galactose, mannose, rhamnose and xylose) were analyzed by RP-HPLC-DAD method. Meanwhile, mannitol and fructose were calculated based on peak areas obtained by HPIEC-RID and corresponding data obtained by RP-HPLC-DAD. Monosaccharide compositions of CDSC were analyzed, nine kinds of monosacchardes (glucose, arabinose, galctose, galacturonic acid, mannose, rhamnose, xylose, fructose and mannitol) in were quantified.Background: Cistanche deserticola Y. C. Ma, is a perennial holoparasite and mainly distributed in the desert region of northwestern China. It reported that C. deserticola soluble carbohydrate (CDSC) could prompt lymphocyte proliferation, modulate immunity, inhibit type 2 diabetes, anti-inflammation. For these reasons, it is very appealing to researchers to have a convenient method for monosaccharides analysis of the CDSC. However, previous researches have neglected the faultiness in monosaccharide composition analysis. Therefore, in this study we will focus on establishing a quantifying method of monosaccharides possibly found in C. deserticola. Methods: Soluble carbohydrates were hydrolyzed with trifluoroacetic acid, monosaccharides were labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP), separated by HPLC system equipped with C18 column and detected with DAD (RP-HPLC-DAD). RP-HPLC-DAD method was used for quantitative analysis of glucose, arabinose, galacturonic acid, galactose, mannose, rhamnose and xylose. Meanwhile, hydrolyzates were separated by HPIEC system equipped with aminex HPX-87H ion exclusion column and detected by refractive index detector (HPIEC-RID). The quantity of mannitol and fructose can be calculated based on peak areas obtained by HPIEC-RID and corresponding data obtained by RP-HPLC-DAD. Results: Utilizing this method, monosaccharide compositions of CDSC with different molecular weight (CDCSA, CDCS-B, CDCS-C and CDCS-D) were analyzed, nine kinds of monosacchardes (glucose, arabinose, galctose, galacturonic acid, mannose, rhamnose, xylose, fructose and mannitol) in were found. Conclusion: Fructose and mannitol, which are abundant present in C. deserticola, cannot be analysis by normally PMP modify method, therefore, a comprehensive quantitative analysis method was developed in this research. Galacturonic acid was found out in macro-molecular polysaccharides from C.deserticola, which was not reported before. Galactose and galacturonic acid mainly exist in macro-molecular polysaccharide. Fructose and mannitol mainly exist in mini-molecule or exist in free form.
Cistanche deserticola Y. C. Ma, soluble carbohydrates, monosaccharide compositions, high performance liquid chromatography, high-performance ion exclusion chromatography, herbal medicine.
Division of Biorefinery Engineering, State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, 100190, No.1 Bei er tiao, Zhongguancun, Haidian District, Beijing 100190