Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China.
Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China; Shandong Provincial Key Laboratory of Glycoscience and Glycoengineering, Ocean University of China, Qingdao 266003, China.
Carbohydr Polym. 2014 Apr 15;104:23-8. doi: 10.1016/j.carbpol.2014.01.011. Epub 2014 Jan 10.
A reliable high performance liquid chromatography with pre-column derivatization method was developed for the determination of the mannuronic acid (M)/guluronic acid (G) ratio of propylene glycol alginate sodium sulfate (PSS). The hydrolysis conditions of PSS were investigated by four degradation methods based on the degree of destruction of M and G, and the chromatographic separation conditions were also optimized. A satisfactory resolution of M and G was achieved with a KP-C18 column using 0.1 mol/L phosphate buffer (pH 7.0)-acetonitrile (83/17, v/v) as a mobile phase, after PSS was hydrolyzed with 0.1 mol/L sulfuric acid and labeled with 1-phenyl-3-methyl -5-pyrazolone. The M/G ratio of PSS determined by this method was in good accordance with that obtained by the (1)H NMR method with a desulfurization strategy. Our method is rapid, sensitive, accurate and reproducible. The limit of detection was found to be 0.25 μg/mL for M and 0.40 μg/mL for G.
建立了一种可靠的高效液相色谱-柱前衍生化法，用于测定丙烯二醇藻酸钠硫酸酯（PSS）中甘露糖醛酸（M）/古洛糖醛酸（G）的比值。通过基于 M 和 G 破坏程度的四种降解方法研究了 PSS 的水解条件，并优化了色谱分离条件。在 0.1mol/L 磷酸缓冲液（pH7.0）-乙腈（83/17，v/v）作为流动相的 KP-C18 柱上，对 0.1mol/L 硫酸水解后的 PSS 进行衍生化和 1-苯基-3-甲基-5-吡唑啉酮标记后，可实现 M 和 G 的良好分离。用这种方法测定的 PSS 的 M/G 比值与脱硫策略的（1）H NMR 法测定的结果非常吻合。该方法快速、灵敏、准确、重现性好。M 的检测限为 0.25μg/mL，G 的检测限为 0.40μg/mL。
