College of Food Science and Engineering, Ocean University of China, 1299 Sansha Road, Qingdao, 266404, China.
Anal Bioanal Chem. 2024 Jun;416(15):3501-3508. doi: 10.1007/s00216-024-05299-5. Epub 2024 Apr 24.
Alginate is a commercially important polysaccharide composed of mannuronic acid and its C5 differential isomer guluronic acid. Comprehensive research on alginate and alginate lyases requires efficient and precise analytical methods for alginate oligosaccharides. In this research, high-performance anion exchange chromatography (HPAEC) in parallel with pulsed amperometric detection (PAD) and mass spectrometry (MS) was applied to the analysis of oligosaccharides obtained by alginate lyase. By optimizing the chromatographic conditions including mobile phase concentration, flow rate, and elution gradient, the analysis of a single sample could be completed in 30 min. Seven unsaturated alginate oligosaccharides were separated and identified through their analysis time observed with PAD, including all structurally different unsaturated disaccharides and trisaccharides. The quantitative analysis of seven oligosaccharides was performed based on the quantitative capability of PAD. The method exhibited adequate linearity and precision parameters. All the calibration curves showed good linearity at least in the concentration range of 0.002 to 0.1 mg/mL. The HPAEC-PAD/MS method provides a general and efficient online method to analyze alginate oligosaccharides.
藻酸盐是一种由甘露糖醛酸及其 C5 差向异构体古罗糖醛酸组成的具有商业重要性的多糖。藻酸盐和藻酸盐裂解酶的综合研究需要对藻酸盐低聚糖进行高效、精确的分析方法。在这项研究中，采用高效阴离子交换色谱（HPAEC）与脉冲安培检测（PAD）和质谱（MS）相结合的方法分析藻酸盐裂解酶得到的寡糖。通过优化包括流动相浓度、流速和洗脱梯度在内的色谱条件，可在 30 分钟内完成单个样品的分析。通过观察 PAD 检测到的分析时间，用 PAD 分离和鉴定了 7 种不饱和藻酸盐低聚糖，包括所有结构不同的不饱和二糖和三糖。基于 PAD 的定量能力，对 7 种寡糖进行了定量分析。该方法表现出足够的线性和精密度参数。所有校准曲线在至少 0.002 至 0.1mg/mL 的浓度范围内均表现出良好的线性。HPAEC-PAD/MS 方法为分析藻酸盐低聚糖提供了一种通用且高效的在线方法。
