糖功能因子高准确定值方法的研究与应用

doi:10.19586/j.2095-2341.2025.0047

生物技术进展 ›› 2025, Vol. 15 ›› Issue (5): 865-873.DOI: 10.19586/j.2095-2341.2025.0047

糖功能因子高准确定值方法的研究与应用

李晶晶¹(), 孙梓晏², 翟睿³, 朱曼曼³, 赵海波¹, 江游³, 梁亮¹, 赵雨佳¹

^1.北京市计量检测科学研究院，北京 100029
^2.东北师范大学生命科学院，长春 130024
^3.中国计量科学研究院，国家市场监管技术创新中心（质谱），前沿计量科学中心，北京 100029

收稿日期:2025-04-09 接受日期:2025-08-28 出版日期:2025-09-25 发布日期:2025-11-11
作者简介:李晶晶 E-mail: lijingjing@bjjl.cn
基金资助:
国家市场监管技术创新技术中心（质谱）项目(AKYKF2409);北京市市场监督管理局科技计划项目(SJKJXM-2024-005)

Research and Application of High-accurate Quantitative Method for Polysaccharide Functional Factors

Jingjing LI¹(), Ziyan SUN², Rui ZHAI³, Manman ZHU³, Haibo ZHAO¹, You JIANG³, Liang LIANG¹, Yujia ZHAO¹

^1.Beijing Institute of Metrology，Beijing 100029，China
^2.School of Life Science，Northeast Normal University，Changchun 130024，China
^3.Technology Innovation Center of Mass Spectrometry for State Marken Regulation，Center for Advanced Measurement Science，National Institute of Metrology，Beijing 100029，China

Received:2025-04-09 Accepted:2025-08-28 Online:2025-09-25 Published:2025-11-11

摘要/Abstract

摘要：

电喷雾电离质谱（electrospray ionization mass spectrometry，ESI-MS）是一种快速、灵敏、准确的分析技术，可精确定量分子含量，同时提供分子质量和结构信息。糖功能因子是功能性食品制造的基础材料，也是赋予产品特定生理功能的标志性成分，主要由多羟基化合物组成。糖功能因子因质子亲和性能和离子化效率低，而亲水性较强，导致进行ESI-MS分析时灵敏度较差。基于此，利用特异性化学衍生化方法结合色谱-质谱联用（liquid chromatograph mass spectrometer，LC-MS）技术测得葡萄糖单糖在0.001~5 μg范围内线性关系良好，检出限和定量限分别为0.001 μg和0.05 μg。以上结果实现了糖功能因子的精确分析和准确定量，拓展了ESI-MS质谱在生物分子计量中的应用。

关键词: 糖功能因子, 葡萄糖, 计量, 高准确定值

Abstract:

Electrospray mass spectrometry （ESI-MS） is a rapid， sensitive and accurate analytical technique， which can accurately quantify molecular content and provide information on molecular mass and structure. Polysaccharide functional factors are fundamental materials for the manufacturing of functional foods， and are also symbolic components that endows products with specific physiological functions， primarily composed of polyhydroxy compounds. The proton affinity and ionization of polysaccharide functional factors are low， while their hydrophilicity are strong， which leads to poor sensitivity when analyzed by ESI-MS. Based on this， a specific chemical derivatization method was used in combination with liquid chromatography mass spectrometer （LC-MS） technology to measure glucose monosaccharides. The linear relationship was good within the range of 0.001~5 μg， with detection limits and quantification limits of 0.001 μg and 0.05 μg， respectively. The above results have achieved precise analysis and accurate quantification of sugar functional factors， expanding the application of ESI-MS in the metrology of biomoleculars.

Key words: polysaccharide functional factors, glucose, metrology, high-accurate quantitative

中图分类号:

李晶晶, 孙梓晏, 翟睿, 朱曼曼, 赵海波, 江游, 梁亮, 赵雨佳. 糖功能因子高准确定值方法的研究与应用[J]. 生物技术进展, 2025, 15(5): 865-873.

Jingjing LI, Ziyan SUN, Rui ZHAI, Manman ZHU, Haibo ZHAO, You JIANG, Liang LIANG, Yujia ZHAO. Research and Application of High-accurate Quantitative Method for Polysaccharide Functional Factors[J]. Current Biotechnology, 2025, 15(5): 865-873.

图/表 9

表1 多重反应监测模式下的质谱参数

Table 1 MS parameters under multiple reaction monitoring mode

化合物	母离子的质荷比/（m·z ^-¹）	子离子质荷比/（m·z^-1）	裂解电压/V	碰撞能量/V
GLc	337.02	270.91	4 500	500
GLc^*	343.04	227.04	4 500	500

图1 单糖PMP衍生化过程中碱性介质的影响

Fig. 1 The effect of alkaline medium on the derivations process of monosaccharide PMP

图2 单糖PMP衍生化过程中反应温度和时间的影响

Fig. 2 The effect of reaction temperature and time on the derivations process of monosaccharide PMP

图3 HPLC分离过程中色谱柱的影响

Fig. 3 The effect of chromatographic colum on the separation process of HPLC

图4 HPLC分离过程中流动相的影响

Fig. 4 The effect of mobile phase on the separation process of HPLC

图5 HPLC分离过程中流速的影响

Fig. 5 The effect of flow velocity on the separation process of HPLC

图6 Glc经正负离子模式扫描质谱图A:负离子模式；B：正离子模式

Fig. 6 MS scanning with positive and negative ion modes of Glc

图7 Glc同位素内标经正离子模式扫描质谱图

Fig. 7 MS scanning with positive ion modes of Glc isotope internal standard

图8 Glc定值结果

Fig. 8 Glc quantitative resuls

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糖功能因子高准确定值方法的研究与应用

Research and Application of High-accurate Quantitative Method for Polysaccharide Functional Factors

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