中药降解寡糖制备和结构表征的研究进展

doi:10.3724/SP.J.1123.2025.08024

摘要/Abstract

摘要：

中药多糖因具有免疫调节、抗肿瘤等优异生物活性而广受关注，但多糖存在分子质量大、单糖组成复杂、支链结构多样的问题，导致其生物利用度偏低、构效关系难以解析，进而限制了其应用。而寡糖分子质量低、结构相对简单且易于纯化与解析，成为揭示中药多糖构效关系的理想模型，其制备、分离纯化及结构表征是该领域的研究核心。本文以中药降解寡糖为核心研究对象，首先从降解策略层面，系统探讨了化学水解、氧化降解、生物酶解及辅助降解等方法，对比分析了不同方法的降解机理、产物分布特征及适用范围，同时阐述了各类方法对寡糖聚合度、端基及支链结构的改造效应，以及对其生物活性的潜在调控作用；随后针对降解寡糖常用的分离纯化色谱技术（凝胶过滤色谱、离子交换色谱、亲水作用色谱及多孔石墨化炭色谱）展开系统评述，详细介绍了各类色谱技术的分离原理、适用场景及在降解寡糖分离中的具体应用；最后针对单糖组成分析、甲基化分析及核磁共振波谱等寡糖结构表征技术，阐明了不同技术间的互补性与各自的局限性。相信随着中药降解寡糖在分离纯化与结构解析领域的持续突破，以及人工智能与多组学等新技术的深度融合，其构效关系将日趋明朗，在功能性食品开发、新药研发及成果转化领域也将发挥更为重要的作用。

关键词: 中药多糖, 降解寡糖, 结构表征, 分离纯化, 结构-活性关系

Abstract:

Polysaccharides derived from traditional Chinese medicine （TCM） have attracted widespread attention due to their excellent bioactivities， such as immunomodulatory， anti-inflammatory， and anti-tumor effects. However， these polysaccharides exhibit inherent characteristics including large molecular weights， complex monosaccharide compositions， and diverse branched structures. These features result in low bioavailability and difficulties in unraveling their structure-activity relationships （SARs）， thereby imposing significant limitations on their practical applications. In contrast， oligosaccharides possess distinct advantages： they have low molecular weights， relatively simple structures， and are easy to purify and characterize structurally. Owing to these properties， oligosaccharides serve as ideal models for deciphering the SARs of TCM polysaccharides. Consequently， the preparation， separation and purification， and structural characterization of TCM oligosaccharides have become the core research focuses in this field.This paper firstly reviews various degradation strategies from the perspective of degradation approaches. These strategies include chemical hydrolysis， oxidative degradation， enzymatic hydrolysis， and assisted degradation methods. For each method， a comparative analysis of their degradation mechanisms， product distribution characteristics， and applicable scopes is conducted. Meanwhile， it elaborates on the modification effects of different methods on the degree of polymerization （DP）， anomeric configuration， and branched structure of oligosaccharides， as well as their potential regulatory roles in the bioactivities of oligosaccharides. Subsequently， this paper carries out a systematic review on the chromatographic techniques commonly used for the separation and purification of degraded oligosaccharides， including gel filtration chromatography （GFC）， ion-exchange chromatography （IEC）， hydrophilic interaction chromatography （HILIC）， and porous graphitized carbon chromatography （PGC）. For each chromatographic technique， this paper provides a detailed account of its separation principle， applicable scenarios， and specific applications in the separation of degraded oligosaccharides. Finally， regarding the oligosaccharide structural characterization techniques-such as monosaccharide composition analysis， methylation analysis， and nuclear magnetic resonance （NMR） spectroscopy-the paper clarifies the complementarity between different techniques and their respective limitations.In summary， this paper focuses on the application of combined strategies-such as chemical hydrolysis， enzymatic hydrolysis， and physicochemical hybrid methods—in the preparation of degraded oligosaccharides derived from TCM， as well as the roles and key points of separation techniques and characterization techniques in structural elucidation. Looking ahead， the preparation， separation and purification， and structural analysis of TCM oligosaccharides require the further establishment of standardized protocols. Additionally， the integrated application of artificial intelligence and multi-omics technologies should be actively promoted in this research field. It is believed that with the continuous in-depth exploration of degraded oligosaccharides， their SARs will become increasingly explicit. It is anticipated that progress in the purification and structural elucidation of TCM-degraded oligosaccharides， together with the convergence of artificial intelligence and multi-omics approaches， will bring clarity to their structure-activity relationships. This will， in turn， enable their more precise and impactful application in functional food design and pharmaceutical development.

Key words: traditional Chinese medicine polysaccharides, degradated oligosaccharide, structural characterization, separation and purification, structure-activity relationship

中图分类号:

O658

刘俊佚, 郑义, 闫竞宇. 中药降解寡糖制备和结构表征的研究进展[J]. 色谱, 2026, 44(8): 889-899.

LIU Junyi, ZHENG Yi, YAN Jingyu. Research advances in the preparation and structural characterization of depolymerized oligosaccharides from traditional Chinese medicine[J]. Chinese Journal of Chromatography, 2026, 44(8): 889-899.

图/表 3

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Comparison of degradation methods	Acid hydrolysis	Oxidative degradation	Enzymatic hydrolysis	Assisted degradation
Core mechanism	electrophilic attack of H⁺ on the glycosidic oxygen atom	non-specific attack by ROS （e.g.， ·OH） on glycosidic bonds and sugar rings	highly specific recognition and cleavage of target glycosidic bonds	enhancement of mass transfer and reaction efficiency via physical fields or novel solvents
Specificity	non-specific cleavage of all acid-labile bonds	non-specific	highly specific to certain linkage types and sugar units	inherits the specificity of the primary degradation method
Condition mildness	high temperature and strong acid required	relatively mild； special equipment for ozone	typically under mild conditions	can lower the required temperature or time of the core reaction
Product controllability	mixture with a broad DP range， low controllability	preliminary control of degradation extent	can directionally produce oligosaccharides of specific DP	significantly improves the controllability of the core method
Main by-products/effects	5-HMF， furfural， monosaccharides， monosaccharide degradation	uronic acid loss； sugar ring opening； aldehyde/carboxyl group introduction	few side reactions， pure products	generally reduced； milder conditions reduce secondary degradation
Structural destructiveness	dehydration； configuration change due to harsh conditions	hydroxyl oxidation； C-C bond cleavage	perfectly preserves sugar ring structures and configurations	aims to mitigate the destructiveness of the core method
Environmental friendliness	strong acid usage； waste acid streams	H₂O₂ decomposes to H₂O； expensive periodate	green process， mild conditions， no hazardous waste	improved efficiency leads to greener processes
Typical application	initial depolymerization， large-scale pretreatment， sample prep for analysis	producing antioxidant oligosaccharides， structural sequencing， material modification	directed preparation of well-defined oligosaccharides for detailed SAR studies	enhanced efficiency of traditional methods

Comparison of degradation methods	Acid hydrolysis	Oxidative degradation	Enzymatic hydrolysis	Assisted degradation
Core mechanism	electrophilic attack of H⁺ on the glycosidic oxygen atom	non-specific attack by ROS （e.g.， ·OH） on glycosidic bonds and sugar rings	highly specific recognition and cleavage of target glycosidic bonds	enhancement of mass transfer and reaction efficiency via physical fields or novel solvents
Specificity	non-specific cleavage of all acid-labile bonds	non-specific	highly specific to certain linkage types and sugar units	inherits the specificity of the primary degradation method
Condition mildness	high temperature and strong acid required	relatively mild； special equipment for ozone	typically under mild conditions	can lower the required temperature or time of the core reaction
Product controllability	mixture with a broad DP range， low controllability	preliminary control of degradation extent	can directionally produce oligosaccharides of specific DP	significantly improves the controllability of the core method
Main by-products/effects	5-HMF， furfural， monosaccharides， monosaccharide degradation	uronic acid loss； sugar ring opening； aldehyde/carboxyl group introduction	few side reactions， pure products	generally reduced； milder conditions reduce secondary degradation
Structural destructiveness	dehydration； configuration change due to harsh conditions	hydroxyl oxidation； C-C bond cleavage	perfectly preserves sugar ring structures and configurations	aims to mitigate the destructiveness of the core method
Environmental friendliness	strong acid usage； waste acid streams	H₂O₂ decomposes to H₂O； expensive periodate	green process， mild conditions， no hazardous waste	improved efficiency leads to greener processes
Typical application	initial depolymerization， large-scale pretreatment， sample prep for analysis	producing antioxidant oligosaccharides， structural sequencing， material modification	directed preparation of well-defined oligosaccharides for detailed SAR studies	enhanced efficiency of traditional methods

Chromatography technique	Stationary phase	Main advantages	Typical application scenarios	Application cases	Ref.
GPC/SEC	porous particles	enables separation by molecular size/hydrodynamic volume； mild conditions	fractionation by degree of polymerization； desalting； removal of large/small molecules	isolation and purification of oligosaccharides produced by hydrolysis of oat β-glucan	［72］
IEC	charged functional groups （e.g.， DEAE， CM）	high capacity for charged molecules； achieves separation by charge density/properties	separation of acidic/charged oligosaccharides； often serves as the first purification step	separation of hydrolyzed oligosaccharides from Plantago major L.	［73］
HILIC	polar groups	exhibits superior performance for polar compounds； high resolution for isomers	separation of neutral/ polar oligosaccharides and isomers； often coupled with MS	separation of native cello-oligosaccharides and oxidized cello-oligosaccharides produced by cellulose hydrolysis	［74］
PGC	porous graphitized carbon	exceptional resolution for isomers； maintains stability under wide pH range	separation of challenging isomeric oligosaccharides for detailed structural analysis	separation and analysis of oligosaccharides from multiple sources	［75］

Chromatography technique	Stationary phase	Main advantages	Typical application scenarios	Application cases	Ref.
GPC/SEC	porous particles	enables separation by molecular size/hydrodynamic volume； mild conditions	fractionation by degree of polymerization； desalting； removal of large/small molecules	isolation and purification of oligosaccharides produced by hydrolysis of oat β-glucan	［72］
IEC	charged functional groups （e.g.， DEAE， CM）	high capacity for charged molecules； achieves separation by charge density/properties	separation of acidic/charged oligosaccharides； often serves as the first purification step	separation of hydrolyzed oligosaccharides from Plantago major L.	［73］
HILIC	polar groups	exhibits superior performance for polar compounds； high resolution for isomers	separation of neutral/ polar oligosaccharides and isomers； often coupled with MS	separation of native cello-oligosaccharides and oxidized cello-oligosaccharides produced by cellulose hydrolysis	［74］
PGC	porous graphitized carbon	exceptional resolution for isomers； maintains stability under wide pH range	separation of challenging isomeric oligosaccharides for detailed structural analysis	separation and analysis of oligosaccharides from multiple sources	［75］

Technique	Primary analytical scope	Core advantages	Major limitations
Monosaccharide composition analysis	qualitative/quantitative analysis of monosaccharide types and molar ratios	high sensitivity； provides fundamental primary information for structural elucidation	prone to monosaccharide degradation during acid hydrolysis； no glycosidic linkage/sequence information
MS	molecular weight determination； molecular formula inference； sequence/partial linkage elucidation	extremely high sensitivity， minimal sample consumption； detailed fragment ion information； compatibility with LC/GC for analyzingcomplex mixtures	limited distinction of isomeric monosaccharides； difficult linkage differentiation via isobaric fragments； no α/β anomeric configuration information； quantitative accuracy dependent on ionization efficiency
Methylation analysis	glycosidic linkage position/branching point identification； hydroxyl group involvement in linkages	recognized as authoritative for direct linkage determination； clear branching architecture revelation； intuitive results comparable with standard libraries	complex； lengthy； multi-step derivatization； ineffective for acidic/sulfate-containing oligosaccharides； destructive to analytes； no sequence/anomeric configuration information
NMR spectroscopy	complete molecular structure elucidation； α/β anomeric configuration； sugar ring conformation； linkage sequence； branch point location	provides complete solution-phase stereochemical and conformational information	requires high-purity， milligram-scale samples； complex spectrum interpretation due to severe signal overlap