响应面法优化宜昌蛇菰多糖提取工艺及柱前衍生HPLC法分析其单糖组成OA

Objective To optimize the extraction process of polysaccharides from Balanophora henryi Hemsl.and analyze its monosaccharide composition,to provide experimental evidence for the development and utilization of B.Henryi resources.Methods Based on single-factor experiments,Box-Behnken design combined with response surface methodology was em-ployed to optimize the water extraction and alcohol precipitation process,with extraction time,extraction temperature and solid-liquid ratio as independent variables and polysaccharide yield as the response value.The crude polysaccharides were purified by the Sevage method,and after hydrolysis with trifluoroacetic acid(TFA),pre-column derivatization with 1-phe-nyl-3-methyl-5-pyrazolone(PMP)was performed,and high-performance liquid chromatography(HPLC)was used for qualitative and relative quantitative analysis of the nine monosaccharides.Results The optimal extraction conditions were determined as follows:solid-liquid ratio of 1∶15,extraction temperature of 90℃and extraction time of 3 h.Under these conditions,the polysaccharide yield reached 5.88%.Monosaccharide composition analysis indicated that the polysaccha-rides from B.henryi consisted of glucuronic acid,mannose,ribose,N-acetylglucosamine,glucosamine,glucose,galac-tose,xylose and fucose,with a relative molar ratio of 12.22∶4.54∶0.62∶2.01∶1.28∶36.61∶21.04∶10.29∶4.19.Conclusion The extraction process optimized by response surface methodology is stable,and the PMP-HPLC method for monosaccharide analysis is reliable.The established methods are suitable for the extraction and quality control of polysac-charides from B.henryi,which lay a theoretical foundation for further research and development of this natural product.