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Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features. Therefore, quantitatively quality control of polysaccharides based on their chemical characteristics is important for their application in biomedical and functional food sciences. However, polysaccharides are mixed macromolecular compounds that are difficult to isolate and lack standards, making them challenging to quantify directly. In this study, we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium erinaceus polysaccharides from laboratory cultured and different regions of China. Briefly, a polysaccharide from H. erinaceus was digested by β-(1-3)-glucanase, and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid (APTS) and separated by using micellar electrokinetic chromatography (MEKC) coupled with laser induced fluorescence (LIF), and quantitatively estimated. MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel electrophoresis (PACE), and provided great peak capacity between oligosaccharides with polymerization degree of 2 (DP2) and polymerization degree of 6 (DP6) in a dextran ladder separation. The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RI) showed that 12 h was sufficient for complete digestion of polysaccharides from H. erinaceus. Laminaritriose (DP3) was used as an internal standard for quantification of all the oligosaccharides. The calibration curve for DP3 showed a good linear regression (R2 > 0.9988). The limit of detection (LOD) and limit of quantification (LOQ) values were 0.05 μg/mL and 0.2 μg/mL, respectively. The recovery for DP3 was 87.32 (±0.03)% in the three independent injections. To sum up, this proposed method is helpful for improving the quality control of polysaccharides from H. erinaceus as well as other materials.

Abstract Background Polygonatum cyrtonema, one of origins of Polygonata Rhizoma (HuangJing in Chinese), is traditionally steamed repeatedly before being used as herbal medicine in China. However, there has no standard for steaming of HuangJing. Therefore, a comprehensive study for effects of steam on polysaccharides from Polygonatum cyrtonema based on saccharide mapping, a powerful method developed for polysaccharides analysis, and pharmacological activity are still necessary, which is helpful to explore the effect of steam on the physiochemical and biological activities of its polysaccharides and develop steaming standard of Polygonatum cyrtonema. Methods To explore the effect of steam on physiochemical and biological activities of P. cyrtonema polysaccharides (PCP), six polysaccharides named PCP0, PCP1, PCP2, PCP3, PCP4 and PCP5 were extracted from the herb consecutively steamed for 0–5 times, respectively. Their molecular weight distribution, monosaccharide composition and PACE fingerprints were investigated through HPSEC-MALLS-RID, HPAEC-PAD and saccharide mapping based on polysaccharides analysis by using carbohydrate gel electrophoresis (PACE) and HPTLC, respectively. In addition, their antioxidant ability and immunostimulatory activities on RAW 264.7 cells in terms of NO production and phagocytosis were compared. Results Results suggested that molecular weights could be changed during steam, which increased by first steaming and then decreased with further steaming though all polysaccharides’ molecular weights were 105-107 Da. They all showed irregularly spherical conformation in aqueous solution based on AFM imaging. Their monosaccharide composition and PACE fingerprints were significantly different after steaming, i.e., galactose increased while glucose and mannose decreased, and β-1,4-Galp appeared while β-1,4-Manp increased, after steaming. Steamed PCP significantly increased scavenging activity against ABTS radicals, while PCP0 had the best immunostimulatory effect on RAW 264.7 in terms of NO production and phagocytosis. Conclusions In summary, steam significantly affected the chemical composition and bioactivities of polysaccharides from P. cyrtonema. Considering the balance beneficial effects of steaming on antioxidant and immunopotentiation activities of PCP, 2 times of continuous steam is the optimal choice under the given conditions.

Polysaccharides exhibit multiple pharmacological activities which are closely related to their structural features. Therefore, quantitatively quality control of polysaccharides based on their chemical characteristics is important for their application in biomedical and functional food sciences. However, polysaccharides are mixed macromolecular compounds that are difficult to isolate and lack standards, making them challenging to quantify directly. In this study, we proposed an improved saccharide mapping method based on the release of specific oligosaccharides for the assessment of Hericium erinaceus polysaccharides from laboratory cultured and different regions of China. Briefly, a polysaccharide from H. erinaceus was digested by β -(1-3)-glucanase, and the released specific oligosaccharides were labeled with 8-aminopyrene-1,3,6-trisulfonic-acid (APTS) and separated by using micellar electrokinetic chromatography (MEKC) coupled with laser induced fluorescence (LIF), and quantitatively estimated. MEKC presented higher resolution compared to polysaccharide analysis using carbohydrate gel electrophoresis (PACE), and provided great peak capacity between oligosaccharides with polymerization degree of 2 (DP2) and polymerization degree of 6 (DP6) in a dextran ladder separation. The results of high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RI) showed that 12 h was sufficient for complete digestion of polysaccharides from H. erinaceus. Laminaritriose (DP3) was used as an internal standard for quantification of all the oligosaccharides. The calibration curve for DP3 showed a good linear regression (R 2 > 0.9988). The limit of detection (LOD) and limit of quantification (LOQ) values were 0.05 μg/mL and 0.2 μg/mL, respectively. The recovery for DP3 was 87.32 (±0.03)% in the three independent injections. To sum up, this proposed method is helpful for improving the quality control of polysaccharides from H. erinaceus as well as other materials. [Display omitted] • Specific oligosaccharides were released from polysaccharides by selected enzyme. • Quantitative saccharide mapping method was firstly developed based on CE-LIF. • This method was superior to PACE, especially the feasibility of quantification. [ABSTRACT FROM AUTHOR]

Grifola frondosa polysaccharides (GFPs) from different regions in China were characterized and compared using HPSEC-MALLS-RID and saccharide mapping based on HPAEC-PAD analysis for achieving and improving its quality control. The results showed that HPSEC chromatograms and molecular weight distributions of GFPs were similar. The average contents of each polysaccharide fraction (Peaks 1, 2, and 3) showed that Peak 3 was the main component and much higher than the other two polysaccharide fractions, which also contained protein. The result of saccharide mapping showed that α-1,4-glycosidic, β-1,4-glycosidic and few β-1,3-glycosidic linkages were existed in GFPs. The similarity result showed that HPAEC-PAD fingerprints of the oligosaccharide fragments after hydrolysis by endoglycosidase were certainly different, especially α-amylase with a mean similar index of only 0.781 ± 0.207. The result of hierarchical cluster analysis (HCA) showed that different batches of GFPs from China can be divided into different clusters. Furthermore, immune-enhancing activity based on RAW 264.7 cells showed significant differences among different GFPs. Based on grey relational analysis (GRA), the fractions of Peak 3 were regarded as the major contributors to its immuno-enhancing activity in GFPs. Overall, the implications from these results were found to be stable, comprehensive, and valid for improving the quality control of GFPs. [ABSTRACT FROM AUTHOR]

Background: Polygonatum cyrtonema, one of origins of Polygonata Rhizoma (HuangJing in Chinese), is traditionally steamed repeatedly before being used as herbal medicine in China. However, there has no standard for steaming of HuangJing. Therefore, a comprehensive study for effects of steam on polysaccharides from Polygonatum cyrtonema based on saccharide mapping, a powerful method developed for polysaccharides analysis, and pharmacological activity are still necessary, which is helpful to explore the effect of steam on the physiochemical and biological activities of its polysaccharides and develop steaming standard of Polygonatum cyrtonema. Methods: To explore the effect of steam on physiochemical and biological activities of P. cyrtonema polysaccharides (PCP), six polysaccharides named PCP0, PCP1, PCP2, PCP3, PCP4 and PCP5 were extracted from the herb consecutively steamed for 0–5 times, respectively. Their molecular weight distribution, monosaccharide composition and PACE fingerprints were investigated through HPSEC-MALLS-RID, HPAEC-PAD and saccharide mapping based on polysaccharides analysis by using carbohydrate gel electrophoresis (PACE) and HPTLC, respectively. In addition, their antioxidant ability and immunostimulatory activities on RAW 264.7 cells in terms of NO production and phagocytosis were compared. Results: Results suggested that molecular weights could be changed during steam, which increased by first steaming and then decreased with further steaming though all polysaccharides' molecular weights were 105-107 Da. They all showed irregularly spherical conformation in aqueous solution based on AFM imaging. Their monosaccharide composition and PACE fingerprints were significantly different after steaming, i.e., galactose increased while glucose and mannose decreased, and β-1,4-Galp appeared while β-1,4-Manp increased, after steaming. Steamed PCP significantly increased scavenging activity against ABTS radicals, while PCP0 had the best immunostimulatory effect on RAW 264.7 in terms of NO production and phagocytosis. Conclusions: In summary, steam significantly affected the chemical composition and bioactivities of polysaccharides from P. cyrtonema. Considering the balance beneficial effects of steaming on antioxidant and immunopotentiation activities of PCP, 2 times of continuous steam is the optimal choice under the given conditions. [ABSTRACT FROM AUTHOR]

Due to the extensive use of xylooligosaccharides (XOS) as functional food ingredients, many inferior goods and even adulterants are generally found in the market, which may pose a health hazard to certain populations. Chromatography method such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) is traditionally applied for the quality analysis of XOS. However, it is time consuming due to the prolonged separation and pre- or post- derivatization procedure. In this study, a fast saccharide mapping method based on matrix-assisted laser desorption/time-of-flight mass spectrometry (MALDI-TOF-MS) was developed for the quality consistency analysis of 22 batches of XOS collected from different manufacturers in China. The time needed for saccharides analysis using MALDI-MS was less than 30 min for one plate, at least 6 times faster than that by the traditional HPTLC chromatography method. In addition, MALDI-MS possessed higher resolution for XOS with DP4-DP7 based on the difference of m/z, which is hardly separated using HPTLC. The results showed that XOS were present only in samples XY01-XY11, samples XY12-XY14 only consisted of hex oligosaccharides, and samples XY15-XY22 were free of oligosaccharides. These indicate that the quality consistency of XOS products in the China market was poor, which should be carefully investigated.

Abstract Brown seaweed polysaccharides (BSPs) are one of the primary active components from brown seaweed that has a range of pharmaceutical and biomedical applications. However, the quality control of BSPs is a challenge due to their complicated structure and macromolecule. In this study, saccharide mapping based on high-performance liquid chromatography (HPLC), multi-angle laser light scattering, viscometer, and refractive index detector (HPSEC-MALLS-Vis-RID), and Fourier transform infrared (FT-IR) were used to discriminate the polysaccharides from nine different species of brown algae (BA1-9). The results showed that BSPs were composed of β-D-glucans and β-1,3−1,4-glucan linkages. The molecular weight, radius of gyration, and intrinsic viscosity of BSPs were ranging from 1.718 × 105 Da to 6.630 × 105 Da, 30.2 nm to 51.5 nm, and 360.99 mL/g to 865.52 mL/g, respectively. Moreover, α values of BSPs were in the range of 0.635 to 0.971, which indicated a rigid rod chain conformation. The antioxidant activities of BSPs exhibited substantial radical scavenging activities against DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2, 2’-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) radicals, which indicated that the use of BSPs might be a potential approach for antioxidant supplements. Thus, this study gives insights about the structure-function relationship of BSPs, which will be beneficial to improve the quality of polysaccharides derived from marine algae.

Saccharide mapping was a promising scheme to unveil the mystery of polysaccharide structure by analysis of the fragments generated from polysaccharide decomposition process. However, saccharide mapping was not widely applied in the polysaccharide analysis for lacking of systematic introduction. In this review, a detailed description of the establishment process of saccharide mapping, the pros and cons of downstream technologies, an overview of the application of saccharide mapping, and practical strategies were summarized. With the updating of the available downstream technologies, saccharide mapping had been expanding its scope of application to various kinds of polysaccharides. The process of saccharide mapping analysis included polysaccharides degradation and hydrolysates analysis, and the degradation process was no longer limited to acid hydrolysis. Some downstream technologies were convenient for rapid qualitative analysis, while others could achieve quantitative analysis. For the more detailed structure information could be provided by saccharide mapping, it was possible to improve the quality control of polysaccharides during preparation and application. This review filled the blank of basic information about saccharide mapping and was helpful for the establishment of a professional workflow for the saccharide mapping application to promote the deep study of polysaccharide structure. [Display omitted] • The detailed structure information was crucial for the deep study and application of polysaccharides. • Saccharide mapping provided a professional workflow for the characterization and identification of polysaccharides. • The methods of obtaining oligosaccharides and several excellent downstream technologies were assessed. • Application scenarios and strategies for saccharide mapping were provided. [ABSTRACT FROM AUTHOR]

Due to the extensive use of xylooligosaccharides (XOS) as functional food ingredients, many inferior goods and even adulterants are generally found in the market, which may pose a health hazard to certain populations. Chromatography method such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) is traditionally applied for the quality analysis of XOS. However, it is time consuming due to the prolonged separation and pre- or post- derivatization procedure. In this study, a fast saccharide mapping method based on matrix-assisted laser desorption/time-of-flight mass spectrometry (MALDI-TOF-MS) was developed for the quality consistency analysis of 22 batches of XOS collected from different manufacturers in China. The time needed for saccharides analysis using MALDI-MS was less than 30 min for one plate, at least 6 times faster than that by the traditional HPTLC chromatography method. In addition, MALDI-MS possessed higher resolution for XOS with DP4-DP7 based on the difference of m / z , which is hardly separated using HPTLC. The results showed that XOS were present only in samples XY01-XY11, samples XY12-XY14 only consisted of hex oligosaccharides, and samples XY15-XY22 were free of oligosaccharides. These indicate that the quality consistency of XOS products in the China market was poor, which should be carefully investigated. • Fast saccharide mapping method was developed based on MADLI-TOF-MS. • Quality consistency of commercial xylooligosaccharides collected in China was evaluated. • Glycosidic linkage analysis was also used for identification of xylooligosaccharides. • Fifty percent of commercial xylooligosaccharides are mislabeled. [ABSTRACT FROM AUTHOR]

The classical Chinese Medicine prescription, Quanzhenyiqitang (QZYQT), containing seven tonic herbs (Shudi, Dangshen, Maidong, Baizhu, Niuxi, Fuzi, and Wuweizi) is clinically used to treat chronic obstructive pulmonary disease (COPD). Although there are studies on the pharmacological effects of QZYQT, little attention has been paid to its active carbohydrate ingredients. We performed a systematic chemical analysis of the crude glycan isolates from the seven-herb decoction (GI-QZYQT) after confirming its anti-COPD activity. GI-QZYQT could enhance lung function, reduce lung damage, and alleviate inflammatory response in mice with COPD. Moreover, two monosaccharides (fructose and glucose) and six oligosaccharides (sucrose, melibiose, 1-kestose, raffinose, mannotriose, and stachyose), accounting for 40.23 % of GI-QZYQT, were discovered using hydrophilic interaction liquid chromatography-evaporative light-scattering detection. Inulin-type fructan with an average molecular weight of 2112 Da was identified using high-performance gel-permeation chromatography in combination with monosaccharide mapping analysis, accounting for 20.10 % of GI-QZYQT in mass. The comparison study showed that the identified monosaccharides, oligosaccharides, and the inulin-type fructan of GI-QZYQT were mainly derived from herbs of Shudi, Dangshen, Maidong, Baizhu, and Niuxi. These findings provide crucial information on the chemical composition of GI-QZYQT, which is vital for the in-depth understanding of its bioactivity, mechanism, and product development., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

Brown seaweed polysaccharides (BSPs) are one of the primary active components from brown seaweed that has a range of pharmaceutical and biomedical applications. However, the quality control of BSPs is a challenge due to their complicated structure and macromolecule. In this study, saccharide mapping based on high-performance liquid chromatography (HPLC), multi-angle laser light scattering, viscometer, and refractive index detector (HPSEC-MALLS-Vis-RID), and Fourier transform infrared (FT-IR) were used to discriminate the polysaccharides from nine different species of brown algae (BA1-9). The results showed that BSPs were composed of β-D-glucans and β-1,3−1,4-glucan linkages. The molecular weight, radius of gyration, and intrinsic viscosity of BSPs were ranging from 1.718 × 105 Da to 6.630 × 105 Da, 30.2 nm to 51.5 nm, and 360.99 mL/g to 865.52 mL/g, respectively. Moreover, α values of BSPs were in the range of 0.635 to 0.971, which indicated a rigid rod chain conformation. The antioxidant activities of BSPs exhibited substantial radical scavenging activities against DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS (2, 2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) radicals, which indicated that the use of BSPs might be a potential approach for antioxidant supplements. Thus, this study gives insights about the structure-function relationship of BSPs, which will be beneficial to improve the quality of polysaccharides derived from marine algae. [ABSTRACT FROM AUTHOR]

In this report, we present the strategy for the revelation of synergistic effect and elucidation of active fractions from an immunomodulatory complex polysaccharide derived from seven herbs (Lentinula edodes , Ganodorma lucidum , Tremella fuciformis , Chrysanthemum , Lycium barbarum , Codonopsis pilosula and Poria cocos), a formula used as health product in China market, using the combination of HPSEC-MALLS, immunological bioassay and saccharide mapping analysis. The effects of complex polysaccharide and their fractions on RAW 246.7 macrophages demonstrated that the fractions (CD1, CD2, CD3) with molecular weight above 10 kDa exhibited immune activity by directly stimulated NO release and phagocytosis, and induced macrophages to secrete cytokines. Especially, fraction CD2 with molecular weight of 100–1000 kDa showed the strongest bioactivity (EC 50 = 0.19 μg/mL) compared with their individual corresponding herbal polysaccharides fractions due to synergistic effect, which supported the scientific use of Chinese herbal mixture. Moreover, their chemical characters were analyzed by HPSEC-MALLS and saccharide mapping, and the original herbs, including L. edodes , G. lucidum , T. fuciformis and Chrysanthemum , responsible for the immunomodulatory activity were tentatively revealed. Results are beneficial for the quality analysis and formula optimization of complex polysaccharides in both biomedical and functional food field. • Immunomodulatory complex polysaccharide was divided into four fractions. • Chemical features were characterized by HPSEC-MALLS and saccharide mapping. • Active fractions, original herbs responsible for the immune activity were revealed. • Complex polysaccharide shows higher immune activity due to synergistic effect. [ABSTRACT FROM AUTHOR]

Qualitative and quantitative analysis of non-starch polysaccharide (NSP) from Pseudostellaria heterophylla in China (Guizhou, Anhui and Fujian) were performed using high performance size exclusion chromatography coupled with multi-angle laser light scattering and refractive index detector (HPSEC-MALLS-RID), saccharide mapping based on polysaccharide analysis by using carbohydrate gel electrophoresis (PACE) and high performance thin layer chromatography (HPTLC). Results showed that PACE and HPTLC fingerprints of enzymatic hydrolysates of NSP were similar. Moreover, the results indicated that 1,4-β-d-Gal p , 1,5-α-Ara f , 1,4-α-d-GalA p and 1,4-β-d-Glc p linkages existed in NSP. Based on the results of HPSEC-MALLS/RI, the Mw , Rg , and contents of NSP in P. heterophylla were ranging from 4.37 × 10 4 to 1.70 × 10 5 Da, 46.4 to 65.9 nm, and 0.43% to 1.41%, respectively. There were no significant difference ( p >0.05) observed among the three main producing areas, which suggested that polysaccharides could be used as marker for quality control of P. heterophylla . [ABSTRACT FROM AUTHOR]

Background Lycium barbarum polysaccharides (LBPs) are considered to be the major bioactive components in L. barbarum . Due to the considerable health benefits of LBPs, and the significant research interest in LBPs, quality evaluation of LBPs and their related products is imperative for ensuring their efficacy and safety. The bioactivities of polysaccharides are closely associated with their physicochemical properties, which can be easily monitored via chemical techniques. Therefore, analysis of physicochemical properties is a more straightforward and viable approach for the quality control of polysaccharides. Scope and approach In this review, special techniques for the structural characterization of LBPs are summarized and discussed. In addition, quality evaluation approaches of LBPs that have never been emphasized are highlighted herein. Furthermore, industrial applications of LBPs are reviewed and discussed. Key findings and conclusions High-performance anion-exchange chromatography coupled with pulsed amperometric detection is one of the most effective techniques for analysis of both acidic and neutral LBPs. Size-exclusion chromatography coupled with multi-angle laser light scattering and refractive index detection is able to determine absolute molecular weights, chain conformations, and contents of LBPs. Fingerprinting analysis and saccharide mapping analysis are effective approaches for quality evaluation of LBPs. However, the lack of appropriate quality evaluation approaches indicates an unreliable regulation of LBPs and their related products. This reveals the deficiency in the quality control of commercial products. Thus, the development of reliable approaches for pharmacological activity-based quality control of LBPs and their related products must be extensively investigated. [ABSTRACT FROM AUTHOR]

Polysaccharides from 39 batches of cultured Cordyceps militaris fruiting body produced in China were investigated and compared using saccharide mapping, partial acidic/enzymatic (α-amylase and β-glucanase) digestion followed by polysaccharide analysis using carbohydrate gel electrophoresis analysis (PACE). The results showed that 1,4-α-D-glucosidic and 1,4-β-D-glucosidic linkages existed in cultivated C. militaris and natural Cordyceps sinensis. There were relatively high similarities among the polysaccharides from different batches of C. militaris, and also between the polysaccharides from C. militaris and natural C. sinensis, which suggested that C. militaris could, at least partially, substitute natural C. sinensis. Furthermore, the results showed that the nitric oxide released from RAW 264.7 cells, induced by polysaccharides from C. militaris and C. sinensis, correlated with their 1,4-α-D-glucosidic and 1,4-β-D-glucosidic linkages, which is beneficial for better understanding of their structure–bioactivity relationship, and helpful to improve pharmacological activity-based quality control of polysaccharides in C. militaris.

The chain conformation, chemical characters and immunomodulatory activity of polysaccharide from Dendrobium devonianum (DDP) were investigated. Results showed that molecular weights, polydispersity index, radius of gyrations of DDP were 3.99 × 10 5  Da 1.27, 74.1 nm, respectively. By applying the polymer solution theory, the exponent (v) values of z 1/2 = kM w v was calculated as 0.38, which revealed that DDP existed as a globular shape in aqueous solution, and further confirmed by AFM analysis. Furthermore, the main monosaccharide compositions were Man and Glc with the ratio of 29.61:1.00. Indeed, the main glycosidic linkages were β-1,4-Manp, and substituted with acetyl groups at O-2 and O-3 position. Notably, DDP could promote the immune functions of macrophages including NO release and phagocytosis. Thus, DDP could be explored as a natural immune-stimulating agent in the health and functional food area as well as pharmaceutical industries. [ABSTRACT FROM AUTHOR]