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柱前衍生化HPLC法测定玉屏风复合多糖的单糖组成

Analysis of monosaccharide compositions of YuPingFeng Polysaccharides by HPLC pre-column derivatization

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【作者】 王少杰巴娟张勇军邓桦蒲文珺杨鸿

【Author】 WANG Shao-jie;BA Juan;ZHANG Yong-jun;DENG Hua;PU Wen-jun;YANG Hong;College of life science and engineering,Foshan University;

【通讯作者】 杨鸿;

【机构】 佛山科学技术学院生命科学与工程学院

【摘要】 本研究旨在采用HPLC法测定玉屏风复合多糖(Yupingfeng polysaccharides,YPF-P)中的单糖组成。水提醇沉提取,AB-8大孔树脂纯化后的YPF-P样品用三氟乙酸溶液水解成单糖,然后将水解后的YPF-P样品与作为对照品的8种单糖D-甘露糖、鼠李糖、D-葡萄糖、D-核糖、D-半乳糖醛酸、D-半乳糖、阿拉伯糖用1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生化,经Wondasil C18(250 mm×4.6 mm,5μm)色谱柱分离,以磷酸盐缓冲溶液(p H值6.9)-乙腈(85∶15)进行洗脱,柱温40℃;流速1.0 mL/min;进样量20μL检测波长254 nm。结果表明,YPF-P是由D-甘露糖、鼠李糖、D-葡萄糖、D-核糖、D-半乳糖醛酸、D-半乳糖、阿拉伯糖组成的,物质的量比为4.64∶1∶5.88∶2.40∶120∶42.9∶69.5。研究结果表明,该方法灵敏度高、样品用量少、稳定性好、准确可靠,能够很好地分离测定YPF-P的各单糖组分。

【Abstract】 This study aims to analyze the monosaccharide compositions of YuPingFeng Polysaccharides( YPF-P) by HPLC.Yupingfeng polysaccharides were extracted by water and ethyl alcohol and then samples were purified by AB-8 macroporous resin,Yupingfeng polysaccharides were hydrolyzed into monosaccharides with trifluoroacetic acid,and then the derivatized samples and the 7 reference monosaccharides,mannose,rhamnose,glucose,ribose,galactose,glucuronic acid,galactose,and Arabia sugar were biochemcially analyzed using 1-phenyl-3-methyl-5-pyrazolone( PMP). The chromatographic separation was carried out on Wondasil C18( 250 mm × 4. 6 mm,5μm) using phosphoric acid aqueous solution-acetonitrile( p H6. 9) and acetonitrile( 85∶ 15) as mobile phases at a flow rate of 1 mL/min kept at 40℃ through gradient elution. The detection wavelength was 254 nm. The results indicated that Yupingfeng polysaccharides was composed of mannose,rhamnose,glucose,ribose,galactose,uronic acid,galactose and Arabia sugar,and their mole ratio was 4. 64∶ 1∶ 5. 88∶ 2. 40∶ 120∶ 42. 9∶ 69. 5. The results also showed that this method had high sensitivity,low sample consumption,good stability and accuracy and reliabibity method. The method is simple and relatively inexpensive for detecting YuPingFeng Polysaccharides.

【关键词】 YPF-PHPLC柱前衍生化分析
【Key words】 YuPingFeng PolysaccharidesHPLCpre column derivatizationanalysis
【基金】 广东省省级科技计划项目(项目编号2016A020210140)
  • 【文献出处】 中国兽医杂志 ,Chinese Journal of Veterinary Medicine , 编辑部邮箱 ,2018年04期
  • 【分类号】S853.7
  • 【被引频次】5
  • 【下载频次】326
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