Microdetermination of Unsaturated Disaccharides Produced from Chondroitin Sulfates in Rabbit Plasma by High Performance Liquid Chromatography with Fluorometric Detection

Toyoda, Hidenao; Shinomiya, Kazufusa; Yamanashi, Shigeyuki; Koshiishi, Ichiro; Imanari, Toshio

doi:10.2116/analsci.4.381

Cited by 60 publications

(26 citation statements)

References 13 publications

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“…Because 2-cyanoacetamide reacts specifically with reducing saccharides, especially unsaturated disaccharides (13), it is possible to quantify HA, C4S, and C6S concentrations in the joint fluid both accurately and specifically. The detection limit of unsaturated disaccharides with this method was -10 picomoles; thus, only a few microliters of joint fluid was required for the analysis.…”

Section: Discussionmentioning

confidence: 99%

“…HPLC analysis of the unsaturated disaccharides derived from HA and CS in joint fluid was performed according to the method of Toyoda et al (13). The HPLC system used in this study was constructed from 2 pumps (model 880-PU; Japan Spectroscopic Co., Ltd., Tokyo, Japan), an autosampling injector (Model 231; Gilson, Villiers le Bel, France), a stainless steel column packed with propylamine-bound silica gel (YMC gel PA-120), a dry reaction bath (DB-3; Shimamura Instrument Co., Tokyo, Japan), a spectrofluorometer (model 820-FP; Japan Spectroscopic), and an integrator (model 805-GI; Japan Spectroscopic).…”

Section: Methodsmentioning

confidence: 99%

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Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Shinmei

Miyauchi

Machida

et al. 1992

Arthritis & Rheumatism

149

108

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Objective. To investigate the correlation between joint disease and the composition of chondroitin sulfate in the joint fluid, unsaturated disaccharide isomers of chondroitin 4-sulfate (.1di-4S) and chondroitin 6-sulfate (.1di·6S) were measured in joint fluids obtained from patients with osteoarthritis (OA), rheumatoid arthritis (RA), or traumatic arthritis (TA).Methods. These pathologic joint fluids were digested with chondroitinase ABC, and the .1di·4S and .1di·6S produced were determined by high performance liquid chromatography combined with fluorometry.Results. Total content of .1di-4S plus .1di·6S was 71.8 ± 30.0 nmoles/ml (mean ± SD) in OA, 55.4 ± 29.3 nmoles/ml in RA, and 211 ± 149 nmoles/ml in TA joint fluids. The ratio of .1di·6S to .1di·4S was 3.81 ± 0.992 in OA, 1.13 ± 0.527 in RA, and 5.75 ± 2.46 in TA joint fluids. Differences between groups were statistically significant.Conclusion. These results strongly suggest that the levels of chondroitin sulfate isomers and the .1di-6S: .1di·4S ratio in joint fluid reflect the proteoglycan metabolism of joint tissues, particularly of articular cartilage; hence, they could be used to diagnose joint diseases

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Shinmei

Miyauchi

Machida

et al. 1992

Arthritis & Rheumatism

149

108

View full text Add to dashboard Cite

show abstract

“…The determination of unsaturated disaccharides (ΔDi-0S, ΔDi-4S, ΔDi-6S, ΔDi-diSE, ΔDi-diSB, ΔDi-diSD, and ΔDi-triS), prepared from the fractionated CS, was performed on the lyase-digested samples using high-performance liquid chromatography [22,23]. Briefly, chondroitinase digestion was performed using tris-acetate buffer (5 L, 0.2 M, pH 8.0) and 10 L of an aqueous solution containing chondroitinase ABC (50 mIU) and chondroitinase ACII (50 mIU) were added to a 20 L portion of the sample solution and incubated at 37 ∘ C for 3 h. An 8 L portion of this mixture was analyzed by high-performance liquid chromatography.…”

Section: Disaccharide and Composition Analyses Of Fractionatedmentioning

confidence: 99%

Effect of Molecular Sizes of Chondroitin Sulfate on Interaction with L-Selectin

Igarashi

Takeguchi

Sakai

et al. 2013

International Journal of Carbohydrate Chemistry

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Chondroitin sulfate (CS) is a glycosaminoglycan (GAG) side chain of proteoglycans (PGs) which are widely distributed in the extracellular matrix and at cell surface. CS shows a highly structural diversity in not only molecular weight (MW) but sulfonation pattern. CS has been reported to exert anti-inflammatory activity by having effects on cytokine production by helper T cells. In this study, we focused on the structures of CS chains, especially MW of CS, and investigated effect of the different MW of CS on binding affinity with L-selectin and cytokine production by murine splenocytes. Firstly, we fractionated CS by employing gel filtration chromatography and obtained several CS fractions with different MW. Then the interaction between fractionated CS and L-selectin was analyzed by surface plasmon resonance (SPR). Finally, the influence of MW of CS on cytokine production by murine splenocytes was investigated in vitro. The results showed that interferon-gamma production was significantly increased by mouse splenocytes cocultivated with CS. On the contrary, CS inhibited interleukin 5 production by murine splenocytes depending on MW of the cocultivated CS. These results strongly indicate the existence of the optimal molecular size for an anti-inflammatory effect of CS through cytokine production by murine splenocytes.

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“…NMR and Compositional Analysis of CS-The determination of unsaturated disaccharides, prepared from CS, was performed on the lyase-digested samples using high-performance liquid chromatography (⌬Di-0S, 3.9%; ⌬Di-4S, 24.5%; ⌬Di-6S, 53.9%; ⌬Di-diS E , 1.7%; ⌬Di-diS B , 0.6%; ⌬Di-diS D , 15.3%; and ⌬Di-triS, 0.2%) (30,31). 1 H NMR spectroscopy was performed using the conditions previously described by Toida et al (10).…”

Section: Methodsmentioning

confidence: 99%

Chondroitin Sulfate Intake Inhibits the IgE-mediated Allergic Response by Down-regulating Th2 Responses in Mice

Sakai

Akiyama

Sato

et al. 2006

Journal of Biological Chemistry

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Chondroitin sulfate (CS) was administered orally to BALB/c mice immunized intraperitoneally with ovalbumin (OVA) and/or dinitrophenylated OVA. The titers of antigen-specific IgE and IgG1 in mouse sera were determined. The antigen-specific IgE production by mice fed ad libitum with CS was significantly inhibited. We also examined the effect of feeding CS on immediate-type hypersensitivity. One hour after antigen stimulation, the ears of mice fed with CS swelled less than those of the control mice. Furthermore, the rise in serum histamine in the mice fed with CS under active systemic anaphylaxis was significantly lower than that in the controls. We next examined the pattern of cytokine production by splenocytes from mice followed by re-stimulation with OVA in vitro. The splenocytes from the mice fed with CS produced less interleukin (IL)-5, IL-10, and IL-13 than those from the control group. In contrast, the production of interferon-␥ and IL-2 by the splenocytes of mice fed with CS was not significantly different from those in the control mice. In addition, the production of transforming growth factor-␤ from the splenocytes of mice fed with CS was significantly higher than that of the control mice. Furthermore, we showed that the percentages of CD4 ؉ cells, CD8 ؉ cells, and CD4؉ CD25 ؉ cells in the splenocytes of mice fed with CS are significantly higher than those of the control. These findings suggest that oral intake of CS inhibits the specific IgE production and antigen-induced anaphylactic response by up-regulating regulatory T-cell differentiation, followed by down-regulating the Th2 response.

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Microdetermination of Unsaturated Disaccharides Produced from Chondroitin Sulfates in Rabbit Plasma by High Performance Liquid Chromatography with Fluorometric Detection

Cited by 60 publications

References 13 publications

Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Quantitation of chondroitin 4—sulfate and chondroitin 6—sulfate in pathologic joint fluid

Effect of Molecular Sizes of Chondroitin Sulfate on Interaction with L-Selectin

Chondroitin Sulfate Intake Inhibits the IgE-mediated Allergic Response by Down-regulating Th2 Responses in Mice

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