柱前衍生超高效液相色谱法分析太子参多糖中单糖的组成

Analysis of monosaccharide composition of Pseudostellaria heterophylla polysaccharides by pre-column derivatization ultra performance liquid chromatography

摘要:

目的  建立柱前衍生超高效液相色谱（UPLC）法，测定太子参多糖中单糖的组成。方法  采用水提醇沉法提取太子参多糖，2 mol/L硫酸水解后加入1-苯基-3-甲基-5-吡唑啉酮进行衍生化，采用UPLC法测定太子参多糖中单糖的衍生物。采用沃特世C18超高效液相色谱柱（100.0 mm×2.1 mm，1.7μm）色谱柱，以乙腈为流动相A，0.1 mol/L磷酸盐（pH 6.8）缓冲液为流动相B，梯度洗脱，检测波长250 nm。通过聚类分析和主成分分析，对不同产地太子参单糖进行质量评价。结果  太子参多糖由半乳糖、D-甘露糖、鼠李糖、阿拉伯糖、D-无水葡萄糖、D-葡萄糖醛酸、D-半乳糖醛酸7种单糖组成，不同产地太子参多糖中单糖组成稍有差别，通过主成分分析和聚类分析评价，10个不同产地太子参单糖聚成3类，并按照单糖含量对各产地进行排名。结论  柱前衍生UPLC法表明，太子参多糖主要由半乳糖醛酸、半乳糖和阿拉伯糖构成，并且贵州施秉县地区产太子参单糖含量较高。

Abstract:

Objective To establish a pre-column derivation ultra performance liquid chromatography (UPLC) for determining monosaccharide composition in Pseudostellaria heterophylla polysaccharides. Methods The polysaccharides were extracted by hot distilled water, precipitated by alcohol, and hydrolyzed with 2 mol/L sulfuric acid and derived by 1-phenyl-3-methyl-5-pyrazolone (PMP). The analysis of monosaccharide composition of P. heterophylla polysaccharides was carried out by reversed-phase UPLC on a Waters ACQUITY UPLC BEH HILIC C18 column (100.0 mm × 2.1 mm, 1.7 μm) with a mobile phase composed of 0.1 mol/L phosphate (pH 6.8) and Acetonitrile in a gradient elution manner. The detection wavelength was set at 250 nm. Cluster analysis (CA) and principal component analysis (PCA) were used to evaluate P. heterophylla monosaccharide in different producing areas. Results The P. heterophylla polysaccharides were composed of galactose, D-mannose, rhamnose, arabinose, D-glucose, D-glucuronic acid and D-galacturonic acid. Totally 10 batches of P. heterophylla polysaccharides could be classified into three clusters, and the producing areas were sorted based on the content of monosaccharides. Conclusions The UPLC with pre-column derivation shows that P. heterophylla polysaccharides mainly consist of galacturonic acid, galactose and arabinose. P. heterophylla of Shibing County has higher content of monosaccharides.