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正交设计法优化镰形棘豆多糖含量测定方法及单糖组成分析 被引量:2

Optimization of content determination of total polysaccharide with orthogonal design and analysis of monosaccharide components in Oxytropis falcate Bunge
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摘要 目的采用正交试验设计优选苯酚-硫酸法测定镰形棘豆总多糖含量的显色条件,并利用高效液相色谱法分析单糖组分。方法以葡萄糖吸光度为指标,选择苯酚用量、浓硫酸用量、水浴温度和水浴时间为考察因素,采用单因素考察结合L9(34)正交试验设计筛选最佳显色条件。多糖经三氟乙酸水解后,用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生水解产物,再利用高效液相色谱分析单糖。结果最佳显色条件为苯酚0.4 mL、浓硫酸5 mL、温度80℃反应10 min。镰形棘豆多糖中含甘露糖、盐酸氨基葡萄糖、核糖、鼠李糖、葡萄糖醛酸、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖的摩尔比为1.21∶0.16∶0.08∶1.00∶0.35∶2.39∶2.32∶2.17∶1.37。结论优化的显色条件能有效提高镰形棘豆总多糖含量测定效率,建立的柱前衍生化高效液相色谱法可用于多糖中的单糖分析。 Objective To optimize the color conditions to determine the total polysaccharide in Oxytropis falcate Bunge by phenol-sulfuric acid method.The monosaccharide compositions of total polysaccharide were analyzed by HPLC.Methods The absorbance of glucose was used as the index.Through single-factor and orthogonal L9 (34) experiments,experimental conditions such as the volume of phenol,the volume of concentrated sulfuric acid,water bath temperature and water bath time were optimized.In addition,the total polysaccharide was hydrolyzed into monosaccharide by trifluoroacetic acid.The hydrolysate was derived by PMP,and then the PMP derivates of monosaccharides were analyzed by HPLC.Results The optimum colored conditions were obtained when the volume of phenol,the volume of concentrated sulfuric acid,water bath temperature,and water bath time were 0.4 mL,5 mL,80℃,and 10 min,respectively.The mole ratio of mannose,glucosamine hydrochloride,ribose,rhamnolipid,glucouronic acid,galacturonic acid,glucose,galactose,and arabinose in the total polysaccharide of Oxytropis falcate Bunge were 1.21∶0.16∶0.08∶1.00∶0.35∶2.39∶2.32∶2.17∶1.37.Conclusion The modified method can effectively improve the determination efficiency of total polysaccharide content in Oxytropis falcate Bunge,and is accurate and reliable.The establised pre-column derivatization HPLC method can be used for the analysis of monosaccharide components in Oxytropis falcate Bunge.
作者 叶财发 蔡跃辉 倪晓霞 蔡小辉 陈锦珊 YE Cai-fa;CAI Yue-hui;NI Xiao-xia;CAI Xiao-hui;CHEN Jin-shan(Department of Pharmacy,No.909 Hospital of the People's Loberation Army/Southeast Hospital Affilated to Xiamen Univesity,Zhangzhou Fujian 363000)
出处 《中南药学》 CAS 2020年第12期2037-2040,共4页 Central South Pharmacy
基金 原南京军区医学科技创新项目(No.14MS085) 医院青年苗圃项目(No.17Y012,No.18Y010)。
关键词 镰形棘豆 正交设计 总多糖 柱前衍生 高效液相色谱法 单糖 Oxytropis falcate Bunge orthogonal design total polysaccharide pre-column derivatization procedure HPLC monosaccharide
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