枸杞子多糖柱前衍生HPLC指纹图谱分析

Pre-column Derivatives HPLC Fingerprint of Lycii Fructus Polysaccharides

摘要:

目的: 建立枸杞子多糖柱前衍生HPLC指纹图谱.方法: 以三氟乙酸(TFA)水解枸杞子多糖,水解产物加入1-苯基-3-甲基-5-吡唑啉酮(PMP)进行衍生化,采用HPLC测定枸杞子多糖中单糖的衍生物.采用ZORBAX Eclipse XDB-C₁₈色谱柱(4.6 mm×250 mm, 5 μm),流动相0.1 mol ·L^-1磷酸缓冲液(pH 6.7)-乙腈(84.5:15.5),流速0.8 mL ·min^-1,检测波长250 nm,柱温30 ℃.结果: 枸杞子多糖柱前衍生指纹图谱标示出11个共有峰,鉴别了8个共有峰(D-甘露糖,L-鼠李糖,D-葡萄糖醛酸,D-半乳糖醛酸,D-葡萄糖,D-半乳糖,D-木糖,D-阿拉伯糖).10批枸杞子药材指纹图谱相似度>0.99.结论: 该方法简便,分离度高,重复性及稳定性良好,可有效控制枸杞子多糖的质量.

Abstract:

Objective:The aim of this study was to establish a pre-column derivation HPLC method for chromatographic fingerprint of Lycii Fructus Polysaccharide. Method: Lycii Fructus sample was hydrolyzed with Trifluoroacetic Acid(TFA) and derivated by 1-phenyl-3-methyl-5-pyrazolone(PMP). The monosaccharide composition of Lycii Fructus polysaccharide was carried out by reversed-phase technique on a ZORBAX Eclipse XDB-C₁₈ column with a mobile phase composed of 0.1 mol · L^-1 phosphate buffer (pH 6.7) and acetonitrile in the ratio of 84.5: 15.5.The flow rate was set at 0.8 mL · min^-1, the wavelength was set at 250 nm, and the column temperature was at 30 ℃. Result: There were 12 peaks from the fingerprint and 8 peaks were D-Man, L-Rha, D-GlcUA, D-GlcN, D-Glc, D-Gal, D-Xyl, D-Ara. The similarity of the 10 batches of Lycium barbarum samples was more than 0.99. Conclusion: The HPLC method with pre-column derivatization is appropriate for the analysis of monosaccharide composition of Lycii Fructus polysaccharide and the method is simple, quick and accurate.