NMPA Key Laboratory for Quality Research and Evaluation of Traditional Chinese Medicine, Shenzhen Institute for Drug Control, Shenzhen 518057, China.
School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
Molecules. 2024 May 13;29(10):2287. doi: 10.3390/molecules29102287.
With the escalating demand for Astragalus polysaccharides products developed from Radix Astragali (RA), the necessity for quality control of polysaccharides in RA has become increasingly urgent. In this study, a specific method for the simultaneous determination of seven monosaccharides in polysaccharides extracted from Radix Astragali (RA) has been developed and validated using ultra-performance liquid chromatography equipped with an ultraviolet detector (UHPLC-UV) for the first time. The 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatizations were separated on a C18 column (Waters ACQUITY, Milfor, MA, USA, 1.8 µm, 2.1 × 100 mm) using gradient elution with a binary system of 5 mm ammonium formate (0.1% formic acid)-acetonitrile for 24 min. Additionally, seven monosaccharides showed good linear relationships (R, 0.9971-0.9995), adequate precision (RSD < 4.21%), and high recoveries (RSD < 4.70%). The established method was used to analyze 109 batches of RA. Results showed that the Astragalus polysaccharides (APSs) mainly consist of mannose (Man), rhamnose (Rha), glucose (Glu), galactose (Gal), arabinose (Ara), xylose (Xyl); and fucose (Fuc); however, their composition was different among RA samples from different growth patterns, species, growth years, and origins, and the growth mode of RA and the age of wild-simulated RA can be accurately distinguished by principal component analysis (PCA). In addition, the immunological activity of APSs were also evaluated jointly by measurement of the NO release with RAW264.7, with the results showing that APSs have a promoting effect on the release of NO and exhibit a significant correlation with Man, Glu, Xyl, and Fuc contents. Accordingly, the new established monosaccharides analytical method and APS-immune activity determination in this study can provide a reference for quality evaluation and the establishment of quality standards for RA.
随着从黄芪中开发的黄芪多糖产品需求的不断增加，对黄芪中多糖质量控制的必要性变得越来越迫切。本研究首次采用超高效液相色谱-紫外检测（UHPLC-UV）建立并验证了同时测定黄芪多糖中七种单糖的方法。采用 1-苯甲酰-3-甲基-5-吡唑啉酮（PMP）衍生化法，在 C18 柱（Waters ACQUITY，Milfor，MA，USA，1.8 µm，2.1×100 mm）上进行分离，采用 5mm 甲酸铵（0.1%甲酸）-乙腈二元系统在 24 分钟内进行梯度洗脱。此外，七种单糖均表现出良好的线性关系（R，0.9971-0.9995）、适当的精密度（RSD<4.21%）和高回收率（RSD<4.70%）。该方法用于分析了 109 批黄芪。结果表明，黄芪多糖（APSs）主要由甘露糖（Man）、鼠李糖（Rha）、葡萄糖（Glu）、半乳糖（Gal）、阿拉伯糖（Ara）、木糖（Xyl）和岩藻糖（Fuc）组成；但不同来源、不同生长模式、不同种、不同生长年份的黄芪样品中其组成存在差异，通过主成分分析（PCA）可准确区分黄芪的生长方式和野生模拟的生长年龄。此外，还通过 RAW264.7 测定 NO 释放量联合评价了 APSs 的免疫活性，结果表明 APSs 对 NO 的释放有促进作用，与 Man、Glu、Xyl 和 Fuc 含量呈显著相关。因此，本研究中建立的新的单糖分析方法和 APS 免疫活性测定方法可为黄芪的质量评价和质量标准的建立提供参考。
