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
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篇名: 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
TITLE: Content Determination of 6 Kinds of Monosaccharide in Astragalus membranaceus with Different Growth Years by Pre-column Derivatization-HPLC
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SymmetryC18为色谱柱,以磷酸盐缓冲液(pH6.8)-乙腈(84∶16,V/V)为流动相,流速为1.0mL/min,检测波长为245nm,柱温为35℃,进样量为20µL。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、2.35~14.04mg/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、122.37~182.41mg/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(162.71~275.77mg/gvs.87.33~107.70、111.54~167.26mg/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。
ABSTRACT: OBJECTIVE:To analyze and compare th e 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 C 18 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 µL. 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 .
期刊: 2021年第32卷第12期
作者: 余亦婷,皮文霞,谢辉,曹丽娟,李西文,李霞,陆兔林,严国俊
AUTHORS: YU Yiting,PI Wenxia ,XIE Hui,CAO Lijuan ,LI Xiwen ,LI Xia,LU Tulin,YAN Guojun
关键字: 黄芪;多糖;单糖;柱前衍生化;高效液相色谱法
KEYWORDS: Astragalus membra naceus;Polysaccharides;Monosaccharides;Pre-column derivatization ;HPLC
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