Enzymatic-fingerprinting workflow of polysaccharides in Hericium erinaceus fruiting bodies: From HILIC-ESI--MS screening to targeted MIM profiling.

In this study, an uncommon enzymatic-fingerprinting workflow, was proposed for characterization and discrimination of mushroom polysaccharides (MPs) by hydrophilic interaction liquid chromatography-negative electrospray mass spectrometry (HILIC-ESI--MS). Firstly, the HILIC-ESI--MS was used to screen and identify the enzymatic digestion products of MPs using HILIC-Orbitrap based on full scan and MS/MS modes. Secondly, a targeted structural-fingerprinting of polysaccharides (SFP) was built in a multiple-ion monitoring (MIM) mode using the same HILIC separation with a triple quadrupole MS. Thirdly, a case study of polysaccharides in Hericium erinaceus fruiting bodies (HEP) was performed to obtain the expected SFP based on dextranase digestion that allows for visual discrimination of polysaccharides from other five edible mushrooms attributed to Agrocybe cylindracea, Arimillaria mellea, Flammulina velutipes, Pleurotus eryngii, and Lentinula edodes. Furthermore, a major structural backbone of HEP was unveiled by occurrence of → 6(Hex)1 → along with multiple possible substitutions including of terminal GalA, Fuc, acetyl, → 4Hex1 →, and → 3Hex1 →. Finally, the similarity analysis, hierarchical cluster analysis (HCA), and partial least squares discriminant analysis (PLS-DA) were performed to visualize various MPs. As a result, the enzymatic-fingerprinting workflow presents an effective example for quality evaluation of fungi polysaccharides using a SFP strategy.

Structural-fingerprinting of polysaccharides to discern Panax species by means of gas-liquid chromatography and mass spectrometry.

In this paper, a sequential gas-liquid chromatography and mass spectrometry route was proposed for characterization of polysaccharides in Panax ginseng (PG), P. notoginseng (PN), and P. quinquefolius (PQ). Due to the reflection of stepped structure parameters, the resulting integrative profiles were tentatively defined as structural-fingerprinting of polysaccharides (SFP) with monosaccharide compositional fingerprinting (MCF), Smith degradation and non-degradation fingerprinting (SDF and SNF), and oligosaccharide compositional fingerprinting (OCF). The MCF, OCF and SDF did not allow for visual discrimination of the three species due to the high interspecific similarity of PG and PQ, whereas SNF could intuitively distinguish PG, PN, and PQ by the presence or absence of Rha and the peak area ratio of Glc/Gal. Similarity analysis, heatmap analysis and principal component analysis were further performed to discern three Panax species based on SNF data sets. The linear →4)-Hexp-(1 â†’ structures were clearly identified as the common structural backbones in side chains or smooth regions of the main chain in PPG, PPN, and PPQ using HILIC-UHPLC-ESI--MS/MS for characterization of partial acid hydrolyzates. The experimental results displayed that the established SFP approach possesses high comprehensibility as well as satisfactory generalization capability for analysis of plant polysaccharides.

Chromatography and mass spectrometry-based approaches for perception of polysaccharides in wild and cultured fruit bodies of Auricularia auricular-judae.

Auricularia auricular-judae polysaccharides (AAPs) have been accepted as one important biological constituent. Quality analysis of A. auricular-judae is generally difficult due to technical issues in separating and detecting AAPs. Here, we describe a stepped chromatographic and mass spectrometric approach for discrimination and characterization of AAPs. HPSEC-MALLS-RID and GC-MS techniques have earlier been shown to be ineffective in evaluating wild and cultured AAPs based on molecular weight distributions and monosaccharide compositions. Nevertheless, direct ESI--MS oligosaccharide fingerprints coupled with PCA have the remarkable ability to discriminate wild and cultured ones. HILIC-UPLC-ESI--MS has indicated that wild and cultured AAPs have distinct peak number and intensities in mild acid hydrolysates. HILIC-UPLC-ESI--HCD-MS/MS was further applied in deducing interglycosidic linkages and sequence of monosaccharide residues in oligosaccharides. We demonstrate that linear GlcAw → Hexm → GlcAx → Hexn → GlcAy → Hexp → GlcAz → Hexq → (w, x, y, z = 0, 1; m, n, p, q ≥ 0; Hex1 → 6Hex, GlcA1 → 4Hex, Hex1 → 4GlcA) was constructed as the general skeleton of AAPs based on C- and B-type ions as well as cross-ring cleavage A-type series ions. The established workflow has been confirmed to be a feasible approach for comprehensive characterization of polysaccharides in foods and plants.