Deng Yong, Zhao Jing, Li Shaoping
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, 999078, China.
Joint Laboratory of Chinese Herbal Glycoengineering and Testing Technology, University of Macau, Macao SAR, 999078, China.
J Pharm Anal. 2023 Feb;13(2):201-208. doi: 10.1016/j.jpha.2022.11.004. Epub 2022 Nov 24.
Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features. Therefore, quantitatively quality control of polysaccharides based on their chemical characteristics is important for their application in biomedical and functional food sciences. However, polysaccharides are mixed macromolecular compounds that are difficult to isolate and lack standards, making them challenging to quantify directly. In this study, we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of polysaccharides from laboratory cultured and different regions of China. Briefly, a polysaccharide from was digested by -(1-3)-glucanase, and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid (APTS) and separated by using micellar electrokinetic chromatography (MEKC) coupled with laser induced fluorescence (LIF), and quantitatively estimated. MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel electrophoresis (PACE), and provided great peak capacity between oligosaccharides with polymerization degree of 2 (DP2) and polymerization degree of 6 (DP6) in a dextran ladder separation. The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RI) showed that 12 h was sufficient for complete digestion of polysaccharides from . Laminaritriose (DP3) was used as an internal standard for quantification of all the oligosaccharides. The calibration curve for DP3 showed a good linear regression ( > 0.9988). The limit of detection (LOD) and limit of quantification (LOQ) values were 0.05 μg/mL and 0.2 μg/mL, respectively. The recovery for DP3 was 87.32 (±0.03)% in the three independent injections. To sum up, this proposed method is helpful for improving the quality control of polysaccharides from as well as other materials.
多糖具有多种药理活性，这些活性与其结构特征密切相关。因此，基于多糖化学特性的定量质量控制对于其在生物医学和功能性食品科学中的应用至关重要。然而，多糖是难以分离且缺乏标准品的混合大分子化合物，直接对其进行定量具有挑战性。在本研究中，我们提出了一种基于特定寡糖释放的改进糖谱法，用于评估实验室培养的以及来自中国不同地区的多糖。简要地说，来自[具体来源未明确]的一种多糖用β-(1-3)-葡聚糖酶消化，释放的特定寡糖用8-氨基芘-1,3,6-三磺酸（APTS）标记，并通过胶束电动色谱（MEKC）结合激光诱导荧光（LIF）进行分离和定量评估。与使用碳水化合物凝胶电泳（PACE）进行多糖分析相比，MEKC具有更高的分辨率，并且在葡聚糖阶梯分离中，在聚合度为2（DP2）和聚合度为6（DP6）的寡糖之间提供了很大的峰容量。高效尺寸排阻色谱结合多角度激光光散射和折射率检测器（HPSEC-MALLS-RI）的结果表明，12小时足以完全消化来自[具体来源未明确]的多糖。以海带三糖（DP3）作为所有寡糖定量的内标。DP3的校准曲线显示出良好的线性回归（r > 0.9988）。检测限（LOD）和定量限（LOQ）值分别为0.05μg/mL和0.2μg/mL。在三次独立进样中，DP3的回收率为87.32（±0.03）%。综上所述，该方法有助于改进[具体来源未明确]以及其他材料中多糖的质量控制。
