高效液相体积排阻色谱法测定稻米支链淀粉链长的相对分子质量分布

doi:10.3864/j.issn.0578-1752.2013.16.020

Abstract

Abstract: 【Objective】The aim of this study is to establish a sensitive high performance size exclusion chromatography with differential refractive index detector (HPSEC-RI) methods for measuring molecular weight distribution of rice amylopectin. 【Method】The proposed method includes three parts: Firstly, separation of amylopectin by butanol precipitation method, Secondly, hydrolysis of rice amylopectin by isoamylase, and Thirdly, qualification of the molecular weight distribution by HPSEC-RI. The proposed parameters of HPSEC-RI are as follows: columns of Shodex SB-G, Shodex SB-803(1 000-100 000) and Shodex SB-802.5(300-10 000) being connected in tandem, column temperature being 40℃, the mobile phase being 0.1 M Tris+0.1 M NaCl (pH=7.40) with a rate of 0.5 mL•min-1, detector temperature being 37℃, sample concentration being 5 mg.mL-1 and the input volume being 20 μL.【Result】The rice amylopectin purified in this study is of high-purity, with the starch content being 99.4%, protein residue being 0.02%, and the peak of rice amylopectin-iodine complex absorption spectrum being 517 nm. The tested rice sample amylopectin chain-length distribution is in the range of DP 2-120. 【Conclusion】The precision and repeatability verification test showed that the proposed method performed well in these aspects including good reproducibility, simple sample preparation, and high efficiency.

Key words: rice amylopectin , HPSEC , molecular weight distribution

YANG Xiao-Yu-12, LIU Zheng-Hui-1, LI Gang-Hua-1, WANG Qiang-Sheng-1, WANG Shao-Hua-1, DING Yan-Feng-1. Determination of Rice Amylopectin Chain-Length Distribution by Using High Performance Size Exclusion Chromatography[J].Scientia Agricultura Sinica, 2013, 46(16): 3488-3495.

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