黄精多糖水解物柱前衍生HPLC指纹图谱

Pre-column Derivatives HPLC Fingerprint of Polygonati Rhizoma Polysaccharide Hydrolysate

摘要:

目的: 建立黄精多糖柱前衍生HPLC指纹图谱. 方法: 以三氟乙酸(TFA)水解黄精多糖,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),检测波长250 nm,柱温30 ℃,流速0.8 mL·min^-1. 结果: 黄精多糖柱前衍生指纹图谱标示出10个共有峰,鉴别了7个共有峰(D-甘露糖, D-半乳糖醛酸, L-鼠李糖, D-半乳糖, D-葡萄糖, D-木糖, D-阿拉伯糖).10批黄精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,可有效控制黄精多糖的质量.

Abstract:

Objective: To establish a pre-column derivation HPLC method of chromatographic fingerprint of Polygonati Rhizoma polysaccharide. Method: Polygonati Rhizoma polysaccharide was hydrolyzed with trifluoroacetic acid and derivated by 1-phenyl-3-methyl-5-pyrazolone (PMP). The monosaccharide composition was separated by reversed-phase technique on a Zorbax Eclipse XDB-C₁₈ column (4.6 mm×250 mm, 5 μm) with a mobile phase composed of 0.1 mol·L^-1phosphate buffer (pH 6.7) and acetonitrile in the ratio of 84.5 : 15.5.The flow rate was 0.8 mL·min^-1. The detection wavelength was set at 250 nm, and column temperature was at 30 ℃. Result: There were 10 common peaks from the fingerprint and 7 peaks were identified as D-mannose, L-rhamnose, D-galacturonic, D-galactose, D-glucose, D-xylose, and D-arabinose. The similarity of 10 batches of Polygonati Rhizoma polysaccharide was more than 0.99. Conclusion: The method is accurate and reproducible, which can be used for the quality control of Polygonati Rhizoma.