PMP柱前衍生化-HPLC法分析不同生长年限黄芪中6种单糖的含量
2021-07-12余亦婷皮文霞谢辉曹丽娟李西文李霞陆兔林严国俊
余亦婷 皮文霞 谢辉 曹丽娟 李西文 李霞 陆兔林 严国俊



摘 要 目的:分析并比较不同生长年限黄芪中6种单糖的含量。方法:取来自3个产地的2~4年生黄芪药材,经水提醇沉、Sevage除蛋白后制得黄芪多糖;将上述多糖经三氟乙酸水解和1-苯基-3-甲基-5-吡唑啉酮衍生化后,采用高效液相色谱法测定其中甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖等6种单糖的含量。以Symmetry C18为色谱柱,以磷酸盐缓冲液(pH 6.8)-乙腈(84 ∶ 16,V/V)为流动相,流速为1.0 mL/min,检测波长为245 nm,柱温为35 ℃,进样量为20 ?L。结果:黄芪多糖中甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖的含量分别为0.50~0.94、0.76~1.60、3.35~7.86、87.33~275.77、1.95~8.96、2.35~14.04 mg/g,2、3、4年生黄芪中上述单糖的总含量分别为98.26~139.92、173.81~295.71、122.37~182.41 mg/g。3年生黄芪中葡萄糖的含量与2、4年生黄芪有显著差异(162.71~275.77 mg/g vs. 87.33~107.70、111.54~167.26 mg/g,P<0.05)。结论:2、3、4年生黄芪中均检出上述6种单糖,其中葡萄糖含量最高,且3年生黄芪中葡萄糖含量显著高于2、4年生黄芪。
关键词 黄芪;多糖;单糖;柱前衍生化;高效液相色谱法
ABSTRACT OBJECTIVE: To analyze and compare the contents of 6 kinds of monosaccharide in Astragalus membranaceus from different growth years. METHODS: 2-4 years old A. membranaceus from three areas were extracted with water extraction and alcohol precipitation, Sevage deproteinization to obtain A. membranaceus polysaccharide. The samples were firstly hydrolyzed with trifluoroacetic acid (TFA) and then derivatized by 1-phenyl-3-methyl-5-pyrazolone(PMP). HPLC analysis was adopted to determine the contents of 6 kinds of monosaccharide as mannose, rhamnose, galacturonic acid, glucose, galactose, arabinose. The determination was performed on Symmetry C18 column with phosphate buffer solution (pH 6.8)-acetonitrile (84 ∶ 16, V/V) as mobile phase at the flow rate of 1.0 mL/min. The detection wavelength was 245 nm, and column temperature was 35 ℃. The sample size was 20 ?L. RESULTS: The contents of mannose, rhamnose, galacturonic acid, glucose, galactose and arabinose were 0.50-0.94, 0.76-1.60, 3.35-7.86, 87.33-275.77, 1.95-8.96, 2.35-14.04 mg/g, respectively. Total contents of monosaccharide from 2, 3, 4 years old A. membranaceus were 98.26-139.92, 173.81-295.71, 122.37-182.41 mg/g, respectively. There was significant difference in the contents of glucose between 3 old years A. membranaceus and 2, 4 old years A. membranaceus (162.71-275.77 mg/g vs. 87.33-107.70, 111.54-167.26 mg/g, P<0.05). CONCLUSIONS: Above 6 monosaccharides are detected in 2, 3, 4 years old A. membranaceus, among which the content of glucose is the highest. The content of glucose in 3 years old A. membranaceus is higher than that in 2 and 4 years old A. membranaceus.
KEYWORDS Astragalus membranaceus;Polysaccharides;Monosaccharides;Pre-column derivatization;HPLC
黃芪为豆科植物蒙古黄芪Astragalus membranaceus(Fisch.)Bge. var. mongholicus(Bge.)Hsiao或膜荚黄芪Astragalus membranaceus(Fisch.)Bge.的干燥根[1],主产于我国山西、内蒙古等地[2]。作为常用传统中药之一,黄芪味甘、性平,具有补气固表、利水生津、托毒排脓、敛疮生肌等功效,临床上主要用于治疗气虚乏力、食少便溏、中气下陷、久泻脱肛、血虚萎黄、内热消渴等症[1,3]。现有研究指出,黄芪具有调节免疫、保护心血管与神经系统、抗肿瘤、护肝等药理作用,其主要化学成分包括黄酮类、皂苷类、多糖类化合物[4-5]。
作为大宗药材之一,黄芪在临床上使用较广,已有研究表明不同生长年限黄芪药材的成分含量存在较大差异,其中黄酮类和皂苷类成分的含量随生长年限的延长而逐年升高,而多糖类成分的含量则变化趋势不一[6-9]。……
