离子色谱HPAEC-PAD法测定杨树细胞壁非纤维素单糖组分
Analysis of Non-Cellulosic Monosaccharide Composition of Poplar Cell Wall by HPAEC-PAD
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摘要: 为建立杨树(Populus spp.)细胞壁中非纤维素单糖的离子交换色谱-脉冲安培检测方法(HPAEC-PAD),通过对分离条件的优化,细胞壁中主要9种单糖可以通过一次进样,达到基线分离的效果。试验采用Dionex CarboPacTM PA100(4 mm×250 mm)分析柱,以氢氧化钠为淋洗液进行梯度洗脱,流速为1.0 mL·min-1,柱温为30℃的色谱条件,测定不同杨树种类细胞壁中单糖组分。结果表明,9种主要细胞壁单糖在0.5~150.0 mg·L-1线性范围内线性关系良好,相关系数R2为0.999 0~0.999 3,方法的重现性(RSD)为1.09%~3.96%,加标回收率为91.32%~109.25%,最低检出限为1.57×10-3~1.41×10-2 mg·L-1。试验结果表明杨树组培苗茎段细胞壁中半乳糖醛酸占比最高,其次为木糖,葡萄糖醛酸占比最低。本研究建立的方法适用范围广,操作简单,可用于杨树及其他物种细胞壁中单糖的检测。Abstract: To establish an ion exchange chromatography-pulsed amperometric detection(HPAEC-PAD) method for non-cellulosic monosaccharides in poplar(Populus spp.) cell walls, the 9 main cell wall monosaccharides could be base-line separated in one run by optimizing the separation conditions. Dionex CarbiPacTM PA100(4 mm×250 mm) with sodium hydroxide analytical column was used for gradient elution at flow rate is 1.0 mL·min-1 and the column temperature of 30 ℃ was used for the determination of monosaccharide fraction in the cell wall of different poplar species. The results showed that the 9 monosaccharides had a good linear relationship in the linear range of 0.5-150.0 mg·L-1, the correlation coefficient R~2 was 0.999 0-0.999 3, the reproducibility(RSD) of the method was 1.09%-3.96%, and the recovery rate by standard addition was 91.32%-109.25%. The detection limit was 1.57×10-3-1.41×10-2 mg·L-1. Galacturonic acid was the highest composition followed by xylose, and the glucuronic acid content was the lowest monosaccharide. The method developed in this study was widely applicable and simple, and could be used for the detection of monosaccharides in the cell walls of poplar and other species.