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目的研究不同因素对三斑海马多糖(HTP)降解的影响,探求适合其单糖组成测定的降解条件。方法通过设计正交实验,采用三氟乙酸(TFA)对HTP进行降解,结合1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生高效液相色谱进行单糖组成分析,比较各实验的降解情况。结果 HTP中主要含有Man、GlcN、Gal和少量的GalN、Glc、GlcA、Ara、Fuc共8种单糖。在考察条件范围内,温度和时间是影响单糖组成分析的主要因素,酸浓度是次要因素。HTP中各单糖的最适降解条件是:氨基糖GlcN和GalN为120℃,6h,4mol/L TFA;中性糖Man、Glc和Gal为110℃,4h,3mol/L TFA;糖醛酸GlcA及Ara、Fuc为100℃,2h,3mol/L TFA。通过对10种单糖标准品的降解也发现氨基糖(GlcN和GalN)较稳定,糖醛酸(GlcA和GalA)稳定性差。结论HTP中不同结构类型的单糖对降解条件的敏感度不同:糖醛酸易降解释放,但也易被破坏;氨基糖较难降解,但降解后较为稳定;中性糖的降解难易和稳定性介于两者之间。采用同一条件降解不能准确地测定HTP的单糖组成,需要针对不同结构类型单糖的性质采用不同的条件进行降解。
Objective To study the effects of different factors on the degradation of three spots of hippocampal polysaccharide (HTP), and to explore the degradation conditions suitable for the determination of monosaccharide composition. Methods Orthogonal experiments were designed and the HTPs were degraded with trifluoroacetic acid (TFA). The monosaccharide composition was determined by PMP column chromatography with 1-phenyl-3-methyl-5-pyrazolone Analyze and compare the degradation of each experiment. Results The main HTP contains Man, GlcN, Gal and a small amount of GalN, Glc, GlcA, Ara, Fuc a total of eight kinds of monosaccharides. Within the range of investigation conditions, temperature and time are the main factors affecting the composition analysis of monosaccharides. Acid concentration is the secondary factor. The optimal conditions for the degradation of each monosaccharide in HTP were as follows: the amino sugar GlcN and GalN were 120 ℃, 6h, 4mol / L TFA; the neutral sugars Man, Glc and Gal were 110 ℃, 4h, 3mol / L TFA; GlcA and Ara, Fuc 100 ℃, 2h, 3mol / L TFA. Amino sugars (GlcN and GalN) were also found to be more stable by degradation of 10 monosaccharide standards, and uronic acids (GlcA and GalA) were less stable. Conclusions Monosaccharides of different structure types in HTP have different sensitivity to degradation conditions. Uronic acids are easily degraded and easily released, but they are also easily destroyed. Amino sugars are more difficult to degrade, but they are stable after degradation. Degradation of neutral sugars is difficult and Stability is in between. Using the same conditions of degradation can not accurately determine the HTP monosaccharide composition, the need for different types of structural properties of different types of sugar using different conditions for degradation.