Structure Elucidation Of Polysaccharides From Pinus Massoniana Pollen And Its Sulfate

Pine pollen is one kind of the pollen which can be used in both food and medicine inChina, and it has functions such as improvement of cardio-cerebral blood vessel,anti-tiredness, antioxidant, liver protection and so on. Pinus massoniana pollen containsprotein, polysaccharide, lipid, vitamin, microelement,nucleic acid and so on.In this research,we studies the extraction, purification, sulfation-technique, and structural analysis of thepolysaccharides from Pinus massoniana pollen with chemical analysis and instrumentanalysis. The main results showd:1. Extraction, Isolation and Purification of Pinus massoniana Pollen polysaccharidesPolysaccharide was extracted from Pinus massoniana pollen by ultrasonic and hot water.The Polysaccharide was purified with column chromatography on Sephacryl S-400HR,naming MPP60-B-â…¡.2. Sulfated modification of Pinus massoniana Pollen polysaccharidesMPP60-B-â…¡ w as chemically modified by chlorosulfonic acid-pyridine method, namingMPPS60-B-â…¡.The content of sulfate radical and degree of substitution in thepolysaecharide were determined by the method of BaSO4turbidimetry. The degree ofsubstitution was1.44.3. Monosaccharide composition of Pinus massoniana Pollen polysaccharidesThe monosaccharide composition of MPP60-B-â…¡ was analyzed by GC after hydrolysisand derivatization. It revealed that MPP60-B-â…¡ composed ofrhamnose, arabinose, fucose,glucose, and galactose in the molar ratios of1.72:5.2:6.66:1:1.16. The content of neutralpolysaccharides was determined by phenol-sulfuric acid method was75.34%. The contentof uronic acid was determined by an improved sulfuric acid-carbazole colorimetry was41.68%. 4. Structure characterization of Pinus massoniana Pollen polysaccharidesThe average molecular weight of MPP60-B-â…¡ and MPPS60-B-â…¡ were about2.08×10~5Da,1.15×10~5Da.The ulrtaviolet spectrum showed that MPP60-B-â…¡ and MPPS60-B-â…¡ had no absorptionin260nm and280nm that means there was no proteins and nuclear acids inMPP60-B-â…¡ and MPPS60-B-â…¡.FT-IR spectroscopy showed that MPP60-B-â…¡ was acidic polysaccharides and theconfiguration of MPP60-B-â…¡ were α-glycosidic and β-glycosidic. MPPS60-B-â…¡ has thecharacteristic absorbance of sulfonic esters.Periodate oxidation and Smith degradation proves that the chain of MPPS60-B-â…¡mainly consisits (1â†'6) glycosidic bond.NMR showed that MPP60-B-â…¡ was acidic polysaccharides and the configuration ofMPP60-B-â…¡ were α-glycosidic and β-glycosidic.Due to the average molecular weight ofMPP60-B-â…¡and the test time of the13C NMR,the continue elucidation of the NMR wasdiscouraged.