CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China; School of Pharmacy, Gansu University of Traditional Chinese Medicine, Lanzhou 730000, China.
CAS Key Laboratory of Chemistry of Northwestern Plant Resources and Key Laboratory for Natural Medicine of Gansu Province, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou 730000, China.
Int J Biol Macromol. 2024 Jan;256(Pt 2):128282. doi: 10.1016/j.ijbiomac.2023.128282. Epub 2023 Nov 24.
The traditional method for isolation and purification of polysaccharides is time-consuming. It often involves toxic solvents that destroy the function and structure of the polysaccharides, thus limiting in-depth research on the essential active ingredient of Lycium barbarum L. Therefore, in this study, high-speed countercurrent chromatography (HSCCC) and aqueous two-phase system (ATPS) were combined for the separation of crude polysaccharides of Lycium barbarum L. (LBPs). Under the optimized HSCCC conditions of PEG1000-KHPO-KHPO-HO (12:10:10:68, w/w), 1.0 g of LBPs-ILs was successfully divided into three fractions (126.0 mg of LBPs-ILs-1, 109.9 mg of LBPs-ILs-2, and 65.4 mg of LBPs-ILs-3). Moreover, ATPS was confirmed as an efficient alternative method of pigment removal for LBPs purification, with significantly better decolorization (97.1 %) than the traditional HO method (88.5 %). Then, the different partitioning behavior of LBPs-ILs in the two-phase system of HSCCC was preliminarily explored, which may be related to the difference in monosaccharide composition of polysaccharides. LBPs-ILs-1 exhibited better hypoglycemic activities than LBPs-ILs-2 and LBPs-ILs-3 in vitro. Therefore, HSCCC, combined with aqueous two-phase system, was an efficient separation and purification method with great potential for separating and purifying active polysaccharides in biological samples.
传统的多糖分离纯化方法耗时耗力。它通常涉及有毒溶剂，这些溶剂会破坏多糖的功能和结构，从而限制了对枸杞 essential 活性成分的深入研究。因此，在本研究中，高速逆流色谱（HSCCC）和双水相体系（ATPS）被结合用于分离枸杞粗多糖（LBPs）。在 PEG1000-KHPO-KHPO-HO（12:10:10:68，w/w）的优化 HSCCC 条件下，成功地将 1.0 g 的 LBPs-ILs 分为三个馏分（126.0 mg 的 LBPs-ILs-1、109.9 mg 的 LBPs-ILs-2 和 65.4 mg 的 LBPs-ILs-3）。此外，ATPS 被证实是一种有效的去除 LBPs 中色素的替代方法，其脱色效果明显优于传统的 HO 法（88.5%）（97.1%）。然后，初步探讨了 LBPs-ILs 在 HSCCC 双水相体系中的不同分配行为，这可能与多糖中单糖组成的差异有关。LBPs-ILs-1 在体外表现出比 LBPs-ILs-2 和 LBPs-ILs-3 更好的降血糖活性。因此，HSCCC 与双水相体系相结合，是一种高效的分离和纯化方法，具有分离和纯化生物样品中活性多糖的巨大潜力。
