THE ANALYSIS AND SEPARATION OF GLUCURONIC AND GALACTURONIC ACIDS BY ION EXCHANGE<sup>1</sup>

Khym, Joseph X.; Doherty, David G.

doi:10.1021/ja01132a536

Cited by 96 publications

(17 citation statements)

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“…Hexosamines were determined by the method of Rondle and Morgan (20) and uronic acid by both the carbazole (Bitter and Muir [21]) and orcinol (Khym and Doherty [22]) techniques using glucuronolactone as a standard. Sulfate was measured by the benzidine technique (Antonopoulos [23]) and hexose (as galactose) by the anthrone reaction (24).…”

Section: Methodsmentioning

confidence: 99%

The Glycosaminoglycans of Normal and Arthritic Cartilage

Mankin¹,

Lippiello²

1971

J. Clin. Invest.

189

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A B S T R A C T The cartilages from the hip joints of 13 normal and 15 osteoarthritic humans were analyzed for glycosaminoglycan content and distribution. The GAGs were separated by elution with CPC on a short cellulose column by the technique of Svejcar and Robertson after digestion of the tissue with pronase and papain.The eluates were identified by a variety of methods including determination of molar ratios, N-acetyl-hexosamine determinations after hyaluronidase treatment and thin-layer chromatography of unhydrolyzed and hydrolyzed GAGs.From the data obtained, it was demonstrated that cartilage from arthritic patients showed a significant increase in the concentration of chondroitin 4-sulfate and a significant decrease in keratan sulfate, with only slight changes in the total amount of GAG present. Calculations of the molar ratios showed variation in the sulfation -with chondroitin 4-sulfate appearing in the "supersulfated" state in the arthritic cartilage.The data lead to speculation regarding the process of osteoarthritis, and it is concluded that the changes seen are more likely to represent an altered pattern of synthesis rather than selective degradation. Since the changes suggest a younger cartilage, a theory is advanced that the chondrocyte responds to the chronic stress of osteoarthritis by modulation to a chondroblastic phase.

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Section: Methodsmentioning

confidence: 99%

The Glycosaminoglycans of Normal and Arthritic Cartilage

Mankin¹,

Lippiello²

1971

J. Clin. Invest.

189

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“…F'roteoglycan extraction of the lightly homogenized cartilage slices was done at 4 O C for 24 h with 15 mL of 4 M guanidine-HCVg tissue, containing protease inhibitors ). The proteoglycan content of representative aliquots was assayed by the orcinol procedure (Khym and Doherty 1952) and by a procedure based on precipitation of proteoglycans with Alcian blue (Bjomsson, personal communication).…”

Section: Total Joint Cartilage Massmentioning

confidence: 99%

Proteoglycan fragments in joint fluid

Dahlberg

Ryd

Lohmander

1992

Acta Orthopaedica Scandinavica

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“…Hexosamine was determined by the method of Boas (1953). Hexuronic acid was determined by the carbazole method of Dische (1947) or Bitter and Muir (1962), and by the orcinol method of Khym and Doherty (1952).…”

Section: Methodsmentioning

confidence: 99%

Extraction of hyaluronic acid from rabbit skin with lanthanum chloride.

Munakata¹,

Yosizawa²

1980

Tohoku J. Exp. Med.

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The defatted rabbit skin was extracted with 0.5 M LaCl3, and the extract was dialyzed exhaustively against distilled water. The precipitate formed during dialysis was dissolved in 28 mM EDTA (ethylenediamine tetraacetate) (pH 7.0). Fr. A was obtained from the solution by precipitation with ethanol in the presence of sodium acetate.The results of gel filtration on Sepharose 4B, electrophoresis on cellulose acetate membrane before and after digestion with Streptomyces hyaluronidase, and analytical data indicated that Fr. A was hyaluronic acid with high molecular weight. The present observation together with previous findings suggest that the binding status of proteoglycans in the skin differs significantly from that in cartilages. hyaluronic acid; rabbit skin; lanthanum chloride Lanthanum chloride was used by Doganges and Schubert (1964) (1978) reported that extraction of cartilage with 0.5 M LaCl3 followed by precipitation on dilution with 9 volumes of water provided a useful initial step for the isolation and separation of proteoglycan and hyaluronic acid. As this method does not involve CsCl density gradient centrifugation, it is more economical both in time and cost than alternative high ionic strength extraction procedure. Recently, Futami et al. (1979) used this method for extraction of carbohydrate-containing substances from non-calcified and calcified portions of bovine costal cartilage, and found that the remodeling of proteoglycan and consumption of glycogen occur on calcification in the cartilage.

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THE ANALYSIS AND SEPARATION OF GLUCURONIC AND GALACTURONIC ACIDS BY ION EXCHANGE¹

Cited by 96 publications

References 1 publication

The Glycosaminoglycans of Normal and Arthritic Cartilage

The Glycosaminoglycans of Normal and Arthritic Cartilage

Proteoglycan fragments in joint fluid

Extraction of hyaluronic acid from rabbit skin with lanthanum chloride.

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THE ANALYSIS AND SEPARATION OF GLUCURONIC AND GALACTURONIC ACIDS BY ION EXCHANGE1

Cited by 96 publications

References 1 publication

The Glycosaminoglycans of Normal and Arthritic Cartilage

The Glycosaminoglycans of Normal and Arthritic Cartilage

Proteoglycan fragments in joint fluid

Extraction of hyaluronic acid from rabbit skin with lanthanum chloride.

Contact Info

Product

Resources

About

THE ANALYSIS AND SEPARATION OF GLUCURONIC AND GALACTURONIC ACIDS BY ION EXCHANGE¹