香菇多糖的PMP-HPLC指纹图谱分析与免疫活性的谱效关系研究  

Fingerprint Chromatography Analysis of Lentinan by PMP-HPLC and its Relationship with Immunoactivity

作　　者：赵秀婷 张燕洁 付萌 李进伟[2] 朱松[2] 范柳萍[2] ZHAO Xiu-ting;ZHANG Yan-jie;FU Meng;LI Jin-wei;ZHU Song;FAN Liu-ping(Infinitus(China)company LTD,Jiangmen,Guangdong 529100,China;School of Food Science and Technology,Jiangnan University,Wuxi,Jiangsu 214122,China)

机构地区：[1]无限极(中国)有限公司,江门529100 [2]江南大学食品学院,无锡214122

出　　处：《粮油食品科技》2021年第3期61-69,共9页Science and Technology of Cereals,Oils and Foods

基　　金：江门市科技计划项目(2020030103440009078)。

摘　　要：研究建立一种柱前1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化液相色谱测定香菇多糖单糖组成的方法,色谱条件为ZORBAX Eclipse XDB-C18液相色谱柱;流动相为0.1 mol/L磷酸盐缓冲液-乙睛(体积比80∶20);柱温30℃;检测器为紫外检测器(245 nm);流速为0.8 mL/min。方法学研究表明:方法准确度高,重现性、稳定性好,各单糖组分在一定的浓度范围内具有良好的线性关系(R2≥0.9985),平均加样回收率在80.6%~91.4%(RSD≤5%),可应用于香菇多糖样品的单糖组成分析。通过对81批次香菇多糖样品的分析,构建了基于多糖单糖组成信息的液相色谱指纹图谱,确定了7个共有特征峰,分别为甘露糖、氨基葡萄糖、核糖、葡萄糖、半乳糖、木糖、岩藻糖,相似度在0.94以上。此外,运用偏最小二乘回归分析(PLS)关联香菇多糖单糖组成与小鼠单核巨噬细胞白血病细胞(RAW264.7细胞)存活率。结果显示,氨基葡萄糖、核糖和葡萄糖与细胞存活率呈正相关,其中前两种单糖的贡献较大。A PMP-HPLC method for the determination of monosaccharide composition of Lentinan was established.Polysaccharide hydrolysates were performed on a Zorbax Eclipse XDB-C18 column,with a mobile phase consisting of 0.1 mol/L phosphate buffer solution-ethyl alcohol(80:20 v/v)at the flow rate of 0.8 m L/min.The detector was ultraviolet detector(245 nm),and the column temperature was 30℃.Methodological studies showed that this method had high accuracy,reproducibility and stability,a good linear relationship in a certain concentration range(R2≥0.9985)and a high sample recovery(80.6%~91.4%,RSD≤5%),so it could be applied to the analysis of monosaccharide composition of Lentinan.Through the analysis of 81 Lentinan samples,a PMP-HPLC fingerprint based on polysaccharide monosaccharide composition was constructed,and seven common characteristic peaks were identified,including fucose,glucosamine,galactose,glucose,mannose and xylose,with the similarity above 0.94.In addition,partial least squares regression(PLS)was used to analyze the correlation of Lentinan monosaccharide composition and the survival rate of RAW264.7 cells.The results showed that the glucosamine,ribose and glucose of Lentinan were positively correlated with the cell survival rate,and the former two had a higher correlation.

关 键 词：香菇多糖 单糖组成 1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生化 高效液相色谱 指纹图谱 免疫活性 

分 类 号：TS201.4[轻工技术与工程—食品科学]