基于串联质谱法和主成分分析的二糖糖苷键的鉴定分析

Identification of glycosidic linkage in disaccharide based on tandem mass spectrometry and principal component analysis

中文关键词:  二糖  糖苷键  高效液相色谱-四极杆飞行时间串联质谱法  主成分分析

英文关键词:disaccharides  glycosidic linkage  high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry  principal component analysis

中文摘要:

      目的 采用高效液相色谱-四极杆飞行时间串联质谱法(high performance liquid chromatography- quadrupole time of flight tandem mass spectrometry, HPLC-Q-TOF-MS/MS)结合主成分分析(principal component analysis, PCA)揭示不同糖苷键二糖的质谱裂解特征, 以用于实际样品中二糖糖苷键的鉴定。方法 采用HPLC-Q-TOF-MS/MS对含有不同糖苷键的二糖标准品进行检测, 采集二糖标准品的MS/MS谱图中离子峰数据, 采用PCA建立二糖糖苷键的预测模型, 并将该模型用于实际低聚半乳糖样品中二糖糖苷键的鉴定分析。结果 PCA结果显示第二主成分(PC2)可区分α和β构型二糖, α构型二糖相比β构型二糖具有较高的[C12H22O11+Na-C6H10O5]+ (m/z 203), 以糖苷键裂解为主; 第一主成分(PC1)可区分α构型二糖中的1→6、1→2糖苷键和1→3、1→4糖苷键, 以及β构型二糖中的1→2糖苷键和1→3、1→4、1→6糖苷键, β(1→2)、α(1→2)、α(1→6)二糖较其他糖苷键二糖在MS/MS谱图上的m/z<185范围内含有较高的碎片离子峰。基于PCA模型可鉴定两种低聚半乳糖样品中的二糖含β(1→3)、β(1→4)、β(1→6)和α(1→3)或α(1→4)键。结论 HPLC-Q-TOF-MS/MS与PCA相结合为实际样品的二糖糖苷键鉴定分析提供了一种简便快速的方法, 可为含低聚糖产品的质量控制和开发提供思路和方法。

英文摘要:

      Objective To explore the mass spectrum cleavage characteristics of disaccharides with different glycosidic linkages to identify the glycosidic linkage of disaccharides in actual samples by high performance liquid chromatography-quadrupole time of flight tandem mass spectrometry (HPLC-Q-TOF-MS/MS) with principal component analysis (PCA). Methods HPLC-Q-TOF-MS/MS was used to analyze commercially available disaccharide standards with different glycosidic linkages. PCA was used to analyze ion peak data in MS/MS spectra of disaccharide standards to establish a prediction model of glycosidic linkages for disaccharides. The model was then used to identify the glycosidic linkages of disaccharides in the galactooligosaccharide samples. Results PCA results revealed that the second principal component (PC2) could distinguish α- and β-disaccharides, α-disaccharides showed higher intensity at [C12H22O11+Na-C6H10O5]+ (m/z 203) than β-disaccharides, with mainly glycosidic linkage cleavage. The first principal component (PC1) could distinguish 1→6, 1→2 with 1→3, 1→4 glycosidic linkage in the α-disaccharides, as well as 1→2 with 1→3, 1→4, 1→6 glycosidic linkage in the β-disaccharides, β(1→2), α(1→2) or α(1→6) showed higher fragment ion peaks in the range of m/z<185 in MS/MS spectra than disaccharides with other linkages. The disaccharides in the 2 galactooligosaccharide samples were identified to contain β(1→3), β(1→4), β(1→6) and α(1→3)/α(1→4) based on the PCA model. Conclusion The combination of HPLC-Q-TOF-MS/MS with PCA provides a simple and rapid method for identifying glycosidic linkage in disaccharides in actual samples, and can provide ideas and methods for the quality control and development of products containing oligosaccharides.