Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides.
Validated HPAEC-PAD Method for the Determination of Fully Deacetylated Chitooligosaccharides.
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用于测定完全脱乙酰壳低聚糖的经过验证的 HPAEC-PAD 方法
DOI：
10.3390/ijms17101699
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发表时间：
2016-10-10
影响因子：
5.6
通讯作者：
Huang Q
中科院分区：
文献类型：
--
作者：
Cao L;Wu J;Li X;Zheng L;Wu M;Liu P;Huang Q
建立了高效阴离子交换色谱脉冲安培检测法（HPAEC-PAD），用于葡萄糖胺（GlcN）1和壳寡糖（COS）（范围为（GlcN）2 ~（GlcN）6）的同时分离和测定。方法的检出限为0.003 ~ 0.016 mg/L（相当于0.4-0.6 pmol），线性范围为0.2 ~ 10 mg/L。采用CarboPac-PA 100色谱柱（4 × 250 mm），以0.2 M氢氧化钠水溶液-水（10：90，v/v）为移动的流动相，流速为0.4 mL/min，进行等度洗脱。回归方程在测试范围内呈良好的线性关系（R2 = 0.9979-0.9995，n = 7）。对质量参数（包括精密度和准确度）进行了充分验证，结果令人满意。完全验证的HPAEC-PAD方法可方便地用于定量商业COS技术浓缩物中的（GlcN）1-6。所建立的方法还用于监测COS技术浓缩物的酸水解，以确保反应条件的优化和（GlcN）1降解的最小化。
An efficient and sensitive analytical method based on high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was established for the simultaneous separation and determination of glucosamine (GlcN)1 and chitooligosaccharides (COS) ranging from (GlcN)2 to (GlcN)6 without prior derivatization. Detection limits were 0.003 to 0.016 mg/L (corresponding to 0.4–0.6 pmol), and the linear range was 0.2 to 10 mg/L. The optimized analysis was carried out on a CarboPac-PA100 analytical column (4 × 250 mm) using isocratic elution with 0.2 M aqueous sodium hydroxide-water mixture (10:90, v/v) as the mobile phase at a 0.4 mL/min flow rate. Regression equations revealed a good linear relationship (R2 = 0.9979–0.9995, n = 7) within the test ranges. Quality parameters, including precision and accuracy, were fully validated and found to be satisfactory. The fully validated HPAEC-PAD method was readily applied for the quantification of (GlcN)1–6 in a commercial COS technical concentrate. The established method was also used to monitor the acid hydrolysis of a COS technical concentrate to ensure optimization of reaction conditions and minimization of (GlcN)1 degradation.
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