Performance Liquid Gel Permeation Chromatography Research Articles

Fucoidan Isolated from Saccharina japonica Inhibits LPS-Induced Inflammation in Macrophages via Blocking NF-κB, MAPK and JAK-STAT Pathways.

Fucoidan has been reported to have a variety of biological activities. However, different algae species, extraction methods, harvesting seasons, and growth regions lead to the structural variation of fucoidan, which would affect the bioactivities of fucoidan. To date, the anti-inflammatory properties and the underlying mechanism of fucoidan from brown alga Saccharina japonica (S. japonica) remain limited. The aims of the present study were to investigate the structure, the anti-inflammatory properties, and the potential molecular mechanisms of fucoidan isolated from S. japonica (SF6) against lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. SF6 was characterized using high performance liquid gel permeation chromatography (HPGPC), Fourier transform infrared spectroscopy (FTIR), and nuclear magnetic resonance spectroscopy (NMR), and observed to be rich in fucose, galactose, and sulfate. Additionally, results showed that SF6 remarkably inhibited LPS-induced production of various inflammatory mediators and pro-inflammation cytokines, including nitric oxide (NO), NO synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-β (IL-β), and interleukin-6 (IL-6). A mechanism study showed that SF6 could effectively inhibit inflammatory responses through blocking LPS-induced inflammation pathways, including nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), and Janus kinase (JAK)-2 and signal transducer and activator of transcription (STAT)-1/3 pathways. These results suggested that SF6 has the potential to be developed as an anti-inflammatory agent applied in functional food.

Optimization extraction, characterization and anticancer activities of polysaccharides from mango pomace

Response surface methodology was used to optimize the extraction conditions for ultrasonic-assisted extraction of polysaccharides from mango pomace. The Optimum extraction conditions consisted of extraction temperature of 74 °C, ultrasonic power of 170 W, extraction time of 100 min, and raw material-to-water ratio of 1:40 g/mL. Under these conditions, the extraction yield was 3.71 ± 0.07%. Three novel polysaccharide fractions, MG-1, MG-2 and MG-3 were purified from the crude polysaccharides by using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of polysaccharide fractions (MPFs) were analyzed by high performance liquid gel permeation chromatography (HPGPC) and HPLC analysis, respectively. The characterizations of MPFs were conducted with FT-IR, 1H NMR and SEM. Furthermore, the anticancer activities of the polysaccharide fractions were also investigated in vitro. Results showed that MG-1, MG-2 and MG-3 exhibited significant anticancer activities against HepG2, MCF-7, A549, HeLa, A2780, HCT-116 and BGC-823 cells in a dose-dependent manner. MPFs were also showed to promote apoptosis as seen in the nuclear morphological examination study using calcein acetyl methoxy methyl easter (calcein-AM) and propidium iodide (PI) staining. This research could serve as a theoretical reference for the efficient utilization of MPFs in biomedical and functional food.

Purification, characterization and hypoglycemic activity of extracellular polysaccharides from Lachnum calyculiforme

Polysaccharides produced by Lachnum have been found as biologically active substances in recent years. Herein, extracellular polysaccharides from Lachnum calyculiforme (LEP) fermenting liquor was obtained by the process of Sevag deproteination, H2O2 decolorization and salting out. LEP-1 was separated and sequentially purified from LEP through DEAE-Cellulose 52 and Sephadex G-100 column chromatography. The results showed that LEP-1 was detected as the homogeneous component with molecular weight of about 445.363kDa by the high performance liquid gel permeation chromatography (HPGPC). Furthermore, we demonstrated that LEP-1 was glucose polymer by GC–MS analysis. IR and NMR spectroscopy (1H, 13C) analysis indicated that the glucose conformation was pyranose-type. Diabetic mice experiments showed that the LEP-1 had a strong hypoglycemic effect in alloxan diabetic mice with dose-dependent relationship.

Molecular weight distribution of carrageenans studied by a combined gel permeation/inductively coupled plasma (GPC/ICP) method

Degraded carrageenan (known as poligeenan, molecular weight: 20kDa to 30kDa) causes ulcerative colitis in experimental animals. In this paper, the molecular weight distributions of 29 samples of food-grade refined carrageenans were studied by high performance liquid gel permeation chromatography (GPC) directly connected to vacuum-ultraviolet inductively coupled plasma-atomic emission spectrometry (ICP) (GPC/ICP) as well as GPC/refractive index (RI) detection. All samples of food-grade carrageenan had a major broad peak of high molecular weight which eluted at around 6.5min in both RI and ICP mode (sulphur and carbon), and each sample of them had no obvious peak of poligeenan (the detection limit was about 5%). The number average molecular weights of these carrageenans ranged from 193 kDa to 324kDa, and the weight average molecular weights ranged from 453kDa to 652kDa based on RI data. Some samples had a few minor peaks which eluted around 10–12min. These peaks came from ionic sulphate, sucrose or glucose. It was considered that if the data-sampling programme was improved, the GPC/ICP system would become a more powerful technique for evaluation of carrageenan samples containing ionic substances and sugar.

Molecular weight distribution of carrageenans studied by a combined gel permeation/inductively coupled plasma (GPC/ICP) method

Degraded carrageenan (known as poligeenan, molecular weight: 20kDa to 30kDa) causes ulcerative colitis in experimental animals. In this paper, the molecular weight distributions of 29 samples of food-grade refined carrageenans were studied by high performance liquid gel permeation chromatography (GPC) directly connected to vacuum-ultraviolet inductively coupled plasma-atomic emission spectrometry (ICP) (GPC/ICP) as well as GPC/refractive index (RI) detection. All samples of food-grade carrageenan had a major broad peak of high molecular weight which eluted at around 6.5min in both RI and ICP mode (sulphur and carbon), and each sample of them had no obvious peak of poligeenan (the detection limit was about 5%). The number average molecular weights of these carrageenans ranged from 193 kDa to 324kDa, and the weight average molecular weights ranged from 453kDa to 652kDa based on RI data. Some samples had a few minor peaks which eluted around 10–12min. These peaks came from ionic sulphate, sucrose or glucose. It was considered that if the data-sampling programme was improved, the GPC/ICP system would become a more powerful technique for evaluation of carrageenan samples containing ionic substances and sugar.

Structures ofCaulerpaCell Wall Microfibril Xylan with Detection ofβ-1,3-Xylooligosaccharides as Revealed by Matrix-assisted Laser Desorption Ionization/Time of Flight/Mass Spectrometry

Since the cell wall microfibril xylans from Caulerpa sp. were not assumed to be constituted from β-1,3-straight chains of homoxylan like Bryopsis maxima but were heteropolysaccharides as glucoxylan or branched structures, identification of its fine structures was desired. Although the results for the linkage analysis were discovered from chemical methods, complementary confirmations were attempted by detection of a series of the β-1,3-xylooligosaccharides prepared as partial acid hydrolysates. Complete separation of the higher oligosaccharide in each was difficult through high performance liquid gel permeation chromatography (GPC), so the composition of oligosaccharides in roughly sub-fractionated samples was detected using matrix-assisted laser desorption ionization/time of flight/mass spectrometry (TOF-MS). By these procedures, it was obvious that cell wall xylan from Caulerpa brachypus was constituted from at least 25 xylose residues of linear oligosaccharides with β-1,3-linkages.Thus the successive me...

Immunofluorescence determination og IgG in cerebrospinal fluid following high performance liquid gel permeation chromatography

A simple technique using high performance liquid gel permeation chromatography for determination of IgG in cerebrolspinal fluid (CSF) is described. Accurate determination of IgG was achieved by precolumn derivation with fluorescein isothiocyanate-conjugated anti-human IgG Fab, a substance that readily yields a fluorescent immune complex. Immune complexes separated as a well-delineated peak in the column void volume, and were measured by the fluorescence of the column eluates (Ex = 490 nm, Em = 520 nm. The results were reproducible with a deviation of less than 2% and a good linear relationship over the range of 2.0–24.5 mg/dl. The IgG concentration assayed by the column method correlated well with values obtained by laser nephelometry.

Immunofluorescence determination of IgG following high performance liquid gel permeation chromatography

The application of high performance liquid gel permeation chromatography (HPL-GPC) and immunofluorescence for the estimation of a protein was studied using human IgG as a model system following separation of the fluorescent immune complex from free fluorescent-labelled antibody on a column of HPL-GPC. Using FITC-labelled anti-human IgG Fab fragment and HPL-GPC, nanogram levels of IgG could be determined within 5 min of mixing the antigen and antiserum. HPL-GPC immunofluorescence analysis is sensitive specific, simple, rapid and highly versatile.