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采用高效阴离子交换色谱-脉冲安培法,建立了不同作物中支链淀粉糖链长分布的分析方法。通过异淀粉酶水解支链淀粉糖分枝点的α-1,6糖苷键,形成不同长度的葡萄糖链。比较了不同色谱柱和不同色谱条件对支链淀粉糖链长分布测定结果的影响,优化了测定方法。以水、0.1 mol/L NaOH和1 mol/L NaAc+0.1 mol/L NaOH溶液为淋洗液进行梯度淋洗,通过CarboPac PA 20阴离子交换柱分离,实现了对聚合度(DP)大于80的支链淀粉糖葡萄糖链的测定。分别选取玉米、木薯、马铃薯与水稻的淀粉进行支链淀粉糖链长分布的研究,并用于其分子构型的判断。实验结果表明该方法简便、检测范围广、重现性良好,连续5次进样,在聚合度小于65时,各峰面积的相对标准偏差小于5%,可为进一步的淀粉糖理化特性研究和品种改良提供有效的实验依据。
The analytical method of amylose chain length distribution in different crops was established by high performance anion exchange chromatography-pulse amperometry. Α-1,6 glucosidic bonds of amylopectin sugar branching points are hydrolyzed by isoamylase to form glucose chains of different lengths. The effect of different chromatographic columns and different chromatographic conditions on the determination of amylose chain length distribution was compared and the determination method was optimized. The gradient elution was carried out with water, 0.1 mol / L NaOH and 1 mol / L NaAc + 0.1 mol / L NaOH as eluent, and separated by CarboPac PA 20 anion exchange column. Determination of amylose, glucose and glucose chains. The starch and amylose content of corn, cassava, potato and rice were selected to study the distribution of amylose chain length, respectively, and used to judge their molecular configuration. The experimental results show that the method is simple, the detection range is wide and the reproducibility is good. The relative standard deviation of each peak area is less than 5% when the polymerization degree is less than 65, which can be used for further study on the starch glyco-chemical properties Variety to provide an effective experimental basis.