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目的:测定土垅大白蚁菌圃多糖中单糖的组成。方法:采用水提醇沉法提取土垅大白蚁菌圃多糖,三氟乙酸水解后,经衍生化试剂1-苯基-3-甲基-吡唑啉酮(PMP)衍生化,高效液相色谱法(HPLC)分析。色谱条件:色谱柱C18(4.6 mm×250 mm,5μm),柱温25℃;流动相为0.05 mol.L-1磷酸二氢钾溶液(pH 6.8)-乙腈,流速1.0 mL.min-1,梯度模式:时间梯度为0→5→10→30→35 min,相应的磷酸盐缓冲溶液梯度为83%→82%→81%→80%→83%;检测波长250 nm;进样体积10μL。结果:土垅大白蚁菌圃多糖中含甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、木糖、阿拉伯糖的摩尔比3.55∶1.00∶1.19∶3.54∶2.54∶4.41∶1.90。结论:该方法具有简单、快速、分离效率高等特点,可用于测定土垅大白蚁菌圃多糖中单糖的组成。
Objective: To determine the composition of monosaccharides in the polysaccharides of soil termites. Methods: Polysaccharides from Termitomyces termites were extracted by water extraction and alcohol precipitation. After the hydrolysis of trifluoroacetic acid, the derivatization reagent 1-phenyl-3-methyl-pyrazolone (PMP) Chromatography (HPLC) analysis. The chromatographic conditions were as follows: column C18 (4.6 mm × 250 mm, 5 μm), column temperature 25 ℃, mobile phase 0.05 mol·L-1 potassium dihydrogen phosphate solution pH 6.8-acetonitrile at a flow rate of 1.0 mL · min- Gradient mode: the time gradient is 0 → 5 → 10 → 30 → 35 min, the corresponding gradient of phosphate buffer solution is 83% → 82% → 81% → 80% → 83%; the detection wavelength is 250 nm and the injection volume is 10 μL. Results: The molar ratio of mannose, rhamnose, galacturonic acid, glucose, galactose, xylose and arabinose in the polysaccharides of Rhodiola spp. Was 3.55:1.00:1.19:3.54:2.54:4.41:1.90. Conclusion: The method has the characteristics of simple, rapid, high separation efficiency and can be used to determine the composition of monosaccharides in the polysaccharides from the soil of the termites termites.