A HILIC-PDA(-MS) profiling approach for the analysis of (methyl esterified) unsaturated galacturonic acid oligosaccharides released from pectins and food by-products

IF 3.4 3区化学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Carbohydrate Research Pub Date : 2026-04-01 Epub Date: 2026-01-12 DOI:10.1016/j.carres.2026.109816

Rebekka Elke Schmidt, Mirko Bunzel

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引用次数: 0

Abstract

Functional and nutritional properties of pectins and pectic oligosaccharides (OS) depend on their structure. A major structural element of pectins are homogalacturonans, which consist of α-1,4-linked galacturonic acids (GalA). Homogalacturonans can be degraded to unsaturated (u)GalA-OS by pectin lyases. To analyze liberated uGalA-OS, a profiling approach based on hydrophilic interaction chromatography (HILIC) with photodiode array (PDA) and optional mass spectrometric (MS) detection was developed. Determination of molar relative response factors (RRF) of de-esterified uGalA-OS (degree of polymerization (DP) = 2–13) in relation to acarbose (internal standard) at 235 nm allows for (semi-)quantitative estimation of (methyl esterified) uGalA-OS and for application in other laboratories. Single quadrupole-MS enables to verify DP and to determine degree of methylation of uGalA-OS. Positions of methyl groups can be tentatively identified by orbitrap-MS. The developed profiling approach can be useful to analyze the specificity of a pectin lyase by analysis of uGalA-OS released from commercial pectin: The pectin lyase applied here appears to prefer a methyl group at cleavage subsite −1, but probably does not require a methyl group at subsite +1. In another application, various methyl esterified uGalA-OS (DP ≤ 12) were identified in enzymatically treated carrot pomace, a pectin-rich food by-product that can potentially be used to enhance nutritional properties of food products.

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用HILIC-PDA（-MS）分析果胶和食品副产品中释放的（甲基酯化）不饱和半乳糖醛酸低聚糖

果胶和果胶寡糖（OS）的功能和营养特性取决于它们的结构。果胶的主要结构元素是半乳糖醛酸，它由α-1,4-链半乳糖醛酸（GalA）组成。高半乳糖醛酸可以通过果胶裂解酶降解为不饱和的(u)GalA-OS。为了分析游离uGalA-OS，建立了一种基于光电二极管阵列（PDA）和可选质谱（MS）检测的亲水性相互作用色谱（HILIC）分析方法。在235 nm下测定去酯化uGalA-OS（聚合度(DP) = 2-13）与阿卡波糖（内标）的摩尔相对响应因子（RRF），可以对（甲基酯化）uGalA-OS进行（半）定量估计，并在其他实验室中应用。单四极杆质谱可以验证DP并确定uGalA-OS的甲基化程度。甲基的位置可以用轨道谱法初步确定。所开发的分析方法可以通过分析商业果胶释放的uGalA-OS来分析果胶裂解酶的特异性：这里应用的果胶裂解酶似乎更倾向于在裂解亚位点- 1上有甲基，但可能不需要在亚位点+1上有甲基。在另一个应用中，在酶处理的胡萝卜渣中鉴定出各种甲基酯化的uGalA-OS (DP≤12)，这是一种富含果胶的食品副产品，可以潜在地用于提高食品的营养特性。

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来源期刊

Carbohydrate Research 化学-生化与分子生物学

CiteScore

5.00

自引率

3.20%

发文量

183

审稿时长

3.6 weeks

期刊介绍： Carbohydrate Research publishes reports of original research in the following areas of carbohydrate science: action of enzymes, analytical chemistry, biochemistry (biosynthesis, degradation, structural and functional biochemistry, conformation, molecular recognition, enzyme mechanisms, carbohydrate-processing enzymes, including glycosidases and glycosyltransferases), chemical synthesis, isolation of natural products, physicochemical studies, reactions and their mechanisms, the study of structures and stereochemistry, and technological aspects. Papers on polysaccharides should have a "molecular" component; that is a paper on new or modified polysaccharides should include structural information and characterization in addition to the usual studies of rheological properties and the like. A paper on a new, naturally occurring polysaccharide should include structural information, defining monosaccharide components and linkage sequence. Papers devoted wholly or partly to X-ray crystallographic studies, or to computational aspects (molecular mechanics or molecular orbital calculations, simulations via molecular dynamics), will be considered if they meet certain criteria. For computational papers the requirements are that the methods used be specified in sufficient detail to permit replication of the results, and that the conclusions be shown to have relevance to experimental observations - the authors'' own data or data from the literature. Specific directions for the presentation of X-ray data are given below under Results and "discussion".