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使用碳酸根修饰的阴离子交换柱对生物质的水提物和水解物中的碳水化合物进行快速分析。

Rapid analysis of carbohydrates in aqueous extracts and hydrolysates of biomass using a carbonate-modified anion-exchange column.

机构信息

Department of Chemistry and Biochemistry, Baylor University, Waco, TX 76798, USA.

出版信息

J Chromatogr A. 2011 Mar 4;1218(9):1236-43. doi: 10.1016/j.chroma.2011.01.002. Epub 2011 Jan 11.

Abstract

Quantitative liquid-chromatography techniques used to characterize carbohydrates present in biomass samples can suffer from long analysis times, limited analyte resolution, poor stability, or a combination of these factors. The current manuscript details a novel procedure enabling resolution of glucose, xylose, arabinose, galactose, mannose, fructose, and sucrose via isocratic elution in less than 5 min. Equivalent conditions also enable analysis of cellobiose and maltose with a minimal increase in chromatographic run time (ca. 3 and 6 min, respectively). Noted chromatographic performance requires that a commercially available anion-exchange column be modified with carbonate prior to analysis. Analytical performance of a modified column was assessed over a 5-day period via repeated analyses of 4 samples, resulting from aqueous extraction or quantitative saccharification of a potential biofuel feedstock (i.e., corn stover or switchgrass). A simple solid phase extraction procedure was utilized to clean up each sample prior to analysis. Analytical accuracy of the extraction protocol was assessed by evaluation of matrix spike recoveries which typically ranged from 84% to 98%. The instrumental variability of measured concentrations in real samples over the 5-day period was generally less than 5% RSD for all detected analytes, independent of sample type. Finally, it is important to note that the modified column exhibited exceptional stability over approximately 800 injections of biofeedstock-based samples. These data demonstrate that a carbonate-modified anion-exchange column can be employed for rapid determination of carbohydrates in biomass samples of lignocellulosic origin.

摘要

用于分析生物质样品中碳水化合物的定量液相色谱技术可能存在分析时间长、分析物分辨率有限、稳定性差或这些因素的组合等问题。本手稿详细介绍了一种新的方法,可在不到 5 分钟的时间内通过等度洗脱来分离葡萄糖、木糖、阿拉伯糖、半乳糖、甘露糖、果糖和蔗糖。在不增加色谱运行时间的情况下(分别约为 3 分钟和 6 分钟),也可以分析纤维二糖和麦芽糖。需要注意的是,这种出色的色谱性能要求在分析前对市售阴离子交换柱进行碳酸盐修饰。通过对来自潜在生物燃料原料(即玉米秸秆或柳枝稷)的水溶液提取或定量糖化的 4 个样品进行重复分析,评估改性柱的分析性能,为期 5 天。在分析之前,每个样品都通过简单的固相萃取程序进行净化。通过评估基质加标回收率来评估提取方案的准确性,回收率通常在 84%至 98%之间。在 5 天的时间内,对实际样品中测量浓度的仪器变异性进行了评估,所有检测到的分析物的相对标准偏差(RSD)通常均小于 5%,与样品类型无关。最后,需要注意的是,改性柱在大约 800 次生物饲料样品的注入中表现出了极好的稳定性。这些数据表明,碳酸盐修饰的阴离子交换柱可用于快速测定木质纤维素生物质样品中的碳水化合物。

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