Ultra-filtration Purification Process And Quality Analysis Of Total Liriope Spicata Polysaccharides

Objective: To establish the preparation process of extraction, ultra-filtration andpurification of total Liriope spicata polysaccharides (TLSP), and quality control ofTLSP.Methods：Preparation of TLSP based on ultra-filtration and purification process.Extraction by hot water, and then removing protein by papain, removing smallmolecules impurity by ultra-filtration, finally removing pigments by DEAE-cellulose52.Process stability and quality analysis. Preliminary studies have completed qualitativeidentification by acid hydrolysis/thin-layer chromatography. Present paper wasdesigned to explore its quality control from two aspects: the stability of ultra-filtrationand purification process and content determination. Analysis of monosaccharidecomposition by gas chromatography and comparative validation analysis of sixreserved ultra-filtration groups. Content Determination used fructose as standard.Using anthrone sulfuric acid/UV-visible colorimetric determination method todetermine the sugar content of TLSP, and using HPGPC-ELSD method (to beconsistent with the latter formulation!) to determine retained monosaccharide andprotein in the polysaccharide.Results:1. Optimum conditions of ultra-filtration and purification process: feedconcentration (extract volume L: herbs weight g)7.5:1, pressure1.2bar, thetangential flow rate1L/min, room temperature at25℃.2. Ultra-filtration yield ofpolysaccharides is about30%, and dialysis yield is about20%, so the yielding ofultra-filtration process was higher than that in the dialysis process, no significantdifference of the purity between these two processes, suggesting that theultra-filtration process efficiency and stability was superior to the dialysis process.3.HPGPC-ELSD results of the samples before and after ultra-filtration suggested thatboth ultra-filtration and dialysis process could effectively remove inorganic salts and small molecular impurities, and the obtained TLSP had no sign of monosaccharide byultra-filtration. UV scan results suggested that ultra-filtration could effectively removeamino acids made by hydrolysis of papain.4. Six samples prepared by ultra-filtrationare composed of fructose and glucose, without other monosaccharides by GC.5.Precision, stability, repeatability, linear correlation, recoveries of the Anthrone-sulfuric acid method for TLSP purity determination was well established and met therequirements of TLSP purity determination.Conclusions: Present experimental results suggested that preparation process ofTLSP by ultra-filtration and purification was stable and reliable; anthrone sulfuricacid/UV-visible colorimetric method could be used as the quality analysis of TLSP.