Structural Characterization and Immunomodulatory Activity of a Novel Polysaccharide From Lycopi Herba

Lycopi Herba has been broadly used as a traditional medicinal herb in Asia due to its ability to strengthen immunity. However, it is still obscure for its material basis and underlying mechanisms. Polysaccharide, as one of the most important components of most natural herbs, usually contributes to the immunomodulatory ability of herbs. Here, we aimed to detect polysaccharides from Lycopi Herba and examine their potential immunomodulatory activity. A novel polysaccharide (LHPW) was extracted from Lycopi Herba and purified by DEAE-52 cellulose chromatography and G-100 sephadex. According to physicochemical methods and monosaccharide composition analysis, LHPW was mainly composed of galactose, glucose, fructose, and arabinose. NMR and methylation analyses indicated that LHPW was a neutral polysaccharide with a backbone containing →3,6)-β-D-Galp-(1→, →4)-β-D-Galp-(1→ and →4)-α-D-Glcp-(1→, with the branches of →1)-β-D-Fruf-(2→ and →6)-α-D-Galp-(1→. Immunological tests indicated that LHPW could activate macrophage RAW264.7 and promote splenocyte proliferation. This study discovered a novel polysaccharide from Lycopi Herba and showed it was a potential immunomodulator.

PCR Instrument-assisted Acidolysis for Monosaccharide Composition Analysis of Serum Glycans

This protocol describes the procedures where a PCR instrument-assisted acidolysis is used for releasing monosaccharides from serum glycans. The monosaccharide composition analysis is subsequently obtained by a HPLC method that separates and quantifies all 1-phenyl-3-methyl-5-pyrazolone (PMP)-labeled monosaccharides in 10 μl serum in 20 minutes. The rapid heating, precise temperature control, and gradient heating properties of PCR instrument provides with consistent acidolysis and derivatization conditions for up to 96 samples simultaneously. The described workflow takes approximately 4–5 h, up to 72 serum samples can be analyzed with one HPLC instrument per day.

Microwave Instrument-assisted Acid Hydrolysis plus HPAEC-PAD for Quantitative Glycan Monosaccharide Composition Analysis of Serum/Plasma Samples

This protocol describes the procedures where our published microwave instrument-assisted acid hydrolysis (MAAH) coupled HPAEC-PAD analysis are optimized for glycan monosaccharide composition analysis of serum/plasma samples. The optimized acid hydrolysis of serum/plasma samples takes only 10 min and 10 μl of acid and 2 μl serum/plasma samples. The monosaccharide composition analysis is subsequently accomplished by HPAEC-PAD analysis. Each step of the experimental procedures has been optimized with repeated tests of monosaccharide standards and serum samples. The described workflow takes approximately 70-90 min, up to 48 serum/plasma samples can be analyzed with one HPAEC-PAD instrument per day.

Ampule-sealed Acidolysis for Monosaccharide Composition Analysis of Serum or Plasma Samples

This protocol describes the procedures where an ampule-sealed acidolysis is used for releasing glycan monosaccharides from serum or plasma samples. The monosaccharide composition analysis is subsequently obtained by a traditional HPLC method that separates and quantifies all 1-phenyl-3-methyl-5-pyrazolone (PMP)-labeled monosaccharides in 10 μl serum in 55 minutes. The described workflow takes approximately 2 days, up to 26 serum samples can be analyzed with one HPLC instrument per day. Each step of the experimental procedures has been optimized with repeated tests of monosaccharide standards and serum samples.

A comparison between Hydrochloric acid and Trifluoroacetic acid in hydrolysis method of exopolysaccharide from Ophiocordyceps sinensis in Monosaccharide composition analysis by GC-FID

The monosaccharide composition is one of the crucial factors affecting the bioactivity of exopolysaccharide (EPS) in Cordyceps species. Therefore, many scientists have studied, analyzed monosaccharide composition and structure of EPS from Cordyceps species, especially Ophiocordyceps sinensis (O. sinensis). This study aimed to compare hydrochloric acid (HCl) with trifluoroacetic acid (TFA) in the EPS hydrolysis process in monosaccharide composition analysis by Gas Chromatography with Flame-Ionization Detection (GC-FID). The hydrolysis is a crucial step in forming the acetyl derivative, which helps the GC-FID technique to have good results in monosaccharide composition analysis. The results showed that hydrolysis with HCl gave a higher hydrolysis efficiency and was more suitable than hydrolysis by TFA in pretreatment to EPS for GC-FID. Hydrolysis results were analyzed through thin-layer chromatography and high-performance liquid chromatography (HPLC), then Acetyl derivatives were produced and finally analyzed by GC-FID to determine the monosaccharide composition of EPS. For EPS hydrolyzed by HCl, the analytical results presented that this sample had 6 kinds of monosaccharides, including rhamnose, arabinose, xylose, mannose, glucose, and galactose; the most monosaccharide was glucose. The EPS hydrolyzed by TFA only detected three kinds of monosaccharides, including mannose, arabinose, and galactose, mainly mannose. The study has set a foundation for further analysis of monosaccharide composition and structure of EPS from O. sinensis.