Australian Centre for Research on Separation Science (ACROSS), School of Natural Sciences, University of Tasmania, GPO Box 252-75, Hobart, Tasmania 7001, Australia; ARC Industrial Transformation Research Hub for Processing Advanced Lignocellulosics (PALS), School of Natural Sciences, University of Tasmania, Hobart, Tasmania 7001, Australia.
ARC Industrial Transformation Research Hub for Processing Advanced Lignocellulosics (PALS), Department of Chemical and Biological Engineering, Monash University, Clayton, Victoria 3800, Australia; Bioresource Processing Research Institute of Australia (BioPRIA), Department of Chemical and Biological Engineering, Monash University, Clayton, Victoria 3800, Australia; Department of Chemical and Biological Engineering, Monash University, Clayton, Victoria, Australia.
J Chromatogr A. 2022 Mar 15;1666:462836. doi: 10.1016/j.chroma.2022.462836. Epub 2022 Jan 22.
High-performance anion-exchange chromatography (HPAEC) coupled with triple quadrupole mass spectrometry (HPAEC-QqQ-MS) was applied to the determination of xylooligosaccharides (XOS) derived from enzymatically hydrolysed commercial xylan from beechwood and the analytical performance and advantages of the method explored. Separation, eluent suppression, electrospray ionisation, and detection options to enhance XOS sensitivity and selectivity were evaluated, delivering a new simple, fast, selective, and sensitive solution for the characterisation of these complex compounds. The method was fully validated in terms of its analytical performance for those XOS for which standards were available, i.e., degree of polymerisation from 1 to 6. The new method was applied to the analysis of xylan hydrolysates obtained by different enzymatic hydrolysis treatments using endo-xylanase from Thermomyces lanuginosus, characterising 25 different XOS and demonstrating the method's utility for future tailoring of enzymatic hydrolysis conditions to obtain desired XOS profiles in such hydrolysates. Linear XOS and 4-O-methyl glucuronic acid (MeGluA) branched XOS were detected by direct injection of the xylan hydrolysates after a simple 10-fold sample dilution and filtration. Identification of XOS detected by HPAEC-QqQ-MS was additionally confirmed using high-resolution orbitrap mass spectrometry (HR-orbitrap-MS). Further, an ultra-sensitive and -selective method was developed by using selected reaction monitoring acquisition mode (SRM), increasing signal-to noise ratio and decreasing the limits of detection, opening future applications to low concentrated sample analysis.
高效阴离子交换色谱（HPAEC）与三重四极杆质谱（HPAEC-QqQ-MS）联用，用于测定酶解商用桦木木聚糖得到的木低聚糖（XOS），并探索了该方法的分析性能和优势。评估了分离、洗脱抑制、电喷雾离子化和检测选项，以提高 XOS 的灵敏度和选择性，为这些复杂化合物的特征提供了一种新的简单、快速、选择性和灵敏的解决方案。
对于那些有标准品的 XOS（聚合度从 1 到 6），该方法在其分析性能方面进行了全面验证。该新方法应用于使用来自嗜热毁丝霉的内切木聚糖酶进行的不同酶解处理得到的木聚糖水解物的分析，鉴定了 25 种不同的 XOS，并证明了该方法在未来根据需要调整酶解条件以获得此类水解物中所需 XOS 谱方面的实用性。
线性 XOS 和 4-O-甲基葡萄糖醛酸（MeGluA）支链 XOS 通过直接注射木聚糖水解物进行检测，水解物经过简单的 10 倍样品稀释和过滤。通过高分辨轨道阱质谱（HR-orbitrap-MS）对 HPAEC-QqQ-MS 检测到的 XOS 进行了进一步确认。此外，通过使用选择反应监测采集模式（SRM）开发了一种超灵敏和选择性方法，提高了信噪比，降低了检测限，为未来对低浓度样品的分析开辟了应用前景。
