Tian Bing-mei, Xie Xiao-mei, Shen Pan-pan, Yang Mo, Zhang Sheng-long, Tang Qing-jiu
Guang Pu Xue Yu Guang Pu Fen Xi. 2015 May;35(5):1331-4.
Chaenomeles speciosa fruits were extracted using water. The extracts were precipitated with 20%~95% (φ) ethanol, respectively. The amount of total polysaccharide was measured with phenol-sulfuric acid method. A method using high-performance size-exclusion chromatography (HPSEC) equipped with multiangle laser-light-scattering photometry (MALLS) and differential refractometry (RI) was presented for determining the molecular weight and molecular weigh distribution. RAW264.7 macrophage were cultured and stimulated with the polysaccharides in vitro and the production of nitric oxide in the cells was determined by the Griess assay. The aim of the study is to determine the amount and the molecular weight of the polysaccharides from Chaenomeles speciosa fruits, and preliminary investigate the immunomodulatory activity, The study provided the basis datas for the further research of Chaenomeles speciosa fruits. , and provided a simple and system method for the research of natural polysaccharide. The ethanol fractional precipitation showed that the order of total polysaccharide content was 95%>80%>40% ≥60%>20%. The results indicated that most polysaccharide from Chaenomeles speciosa fruits might be precipitated when ethanol concentration was up to 95% (T) and the crude polysaccharide purity had risen from 35. 1% to 45. 0% when the concentration of ethanol increased from 20% to 95%. HPSEC-MALLS-RI system showed that all the polysaccharide samples had the similar compositions. They appeared three chromatographic peaks and the retention time were not apparently different. The Mw were 6. 570 X 10(4) g . mol-1 and 1. 393 X 10(4) g . mol-1 respectively, and one less than 10 000 which was failure to obtain accurate values. The molecular weight of the first two polysaccharide distribution index(Mw/Mn)were 1. 336 and 1. 639 respectively. The polysaccharide samples had not exhibited immunomodulatory activity assessed on the basis of nitric oxide production by RAW264. 7 macrophage cells in the experiment.
采用水提取木瓜果实。提取物分别用20%~95%（体积分数）的乙醇沉淀。用苯酚-硫酸法测定总多糖含量。提出了一种使用配备多角度激光散射光度法（MALLS）和示差折光法（RI）的高效尺寸排阻色谱法（HPSEC）来测定分子量和分子量分布的方法。培养RAW264.7巨噬细胞并在体外用多糖刺激，通过Griess法测定细胞中一氧化氮的产生。本研究的目的是测定木瓜果实多糖的含量和分子量，并初步研究其免疫调节活性，为木瓜果实的进一步研究提供基础数据，为天然多糖的研究提供一种简单系统的方法。乙醇分级沉淀表明，总多糖含量顺序为95%>80%>40%≥60%>20%。结果表明，当乙醇浓度达到95%（体积分数）时，木瓜果实中的大部分多糖可能沉淀出来，当乙醇浓度从20%增加到95%时，粗多糖纯度从35.1%提高到45.0%。HPSEC-MALLS-RI系统表明，所有多糖样品组成相似。它们出现三个色谱峰，保留时间无明显差异。分子量分别为6.570×10⁴g·mol⁻¹和1.393×10⁴g·mol⁻¹，其中一个小于10000，未能获得准确值。前两种多糖的分子量分布指数（Mw/Mn）分别为1.336和1.639。在实验中，基于RAW264.7巨噬细胞产生一氧化氮的评估，多糖样品未表现出免疫调节活性。
