目的 采用多种技术手段对黄芪多糖对照品进行标化，以应用于注射用黄芪多糖的质量控制。方法 采用紫外分光光度（UV）法对总糖量进行定量分析，采用HPLC法于250 nm波长处检测1-苯基-3-甲基-5-吡唑啉酮（PMP）衍生化后单糖组成及其量，采用高效凝胶渗透色谱法测定多糖相对分子质量（M_W）及其分布，用薄层色谱法检查其中游离单糖，并采用气相色谱串联质谱法和核磁共振波谱（¹H-NMR）对糖苷键连接位置及构型进行判断。采用多个试验参数对首批黄芪多糖对照品的质量进行控制。结果 此批黄芪多糖对照品由鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖组成，其中葡萄糖为主要成分，且本品中未检测出游离单糖；M_W测定结果显示本品依次出现4个色谱峰，其中主峰（3号峰）M_W为12 531，峰面积比例为83%；总糖量以葡萄糖计为94%；气质联用法测得糖苷键类型主要有8种，其中以1，4-葡萄糖连接为主；核磁确认异头氢为α构型。结论 建立了黄芪多糖对照品标化的方法，其定性鉴别及总糖定量测定结果准确、可靠，可进一步对其在黄芪多糖产品中的质量控制技术进行研究。

Objective A variety of techniques were adopted to establish a standardization method of Astragalus polysaccharides (APs) reference substance. This reference substance was used for quality control of APs for injection. Methods The total sugar content was quantified by UV spectrophotometry ration. The content of monosaccharide was determined by HPLC at detective wavelength of 250 nm after derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP). The high performance size exclusion chromatography method was used for the determination of molecular weight and distribution. TLC was applied to check the free monosaccharide. Then the glycosidic linkage position and configuration were detected by using gas chromatography tandem mass spectrometry and nuclear magnetic resonance spectroscopy (¹H-NMR). A number of experimental parameters were applied to control the quality of APs reference substance. Results The APs was composed of rhamnose (Rha), galactose acid (GalA), glucose (Glc), galactose (Gal), and arabinose (Arb), and Glc was the main component and free monosaccharide were not detected in APs. The result of molecular weight test showed that there were four chromatographic peaks in APs, of which the main peak (peak 3) M_W was 12 531 and the peak area ratio was 83%. The amount of sugar was 94% by UV test in which the glc was calculated as reference substance. There were eight main types of glycosidic bonds detected by GC-MS. Among them, the 1,4-glucose linkage was the main one. The glycosidic bond confirmed by NMR was α configuration. Conclusion The standardization method of APs reference substance has been established in this study. The identification method and determination of sugar amount are accurate and reliable. In the further research, APs reference substance will be used in quality control of APs products.