Journal of Lipid Research, Vol. 9, 319-327, May 1968
Copyright © 1968 by Lipid Research, Inc.

Rapid, sensitive spectrophotometric method for quantitative determination of sulfatides

Edward L. Kean

Departments of Ophthalmology and Biochemistry, Case Western Reserve University School of Medicine, Cleveland, Ohio 44106

A rapid, sensitive spectrophotometric method for the analysis of sulfatides is described. The assay method is based upon the formation of a colored complex between the cationic dye azure A and (anionic) sulfolipids; the complex is extractable by a solution of chloroform-methanol 1:1. With the exception of some of the phospholipids, the reaction is specific for the sulfolipids. Cardiolipin was the most reactive of the non-sulfolipid compounds that were tested. Except for sphingosine and the mono- and dihexose derivatives of sphingosine, a wide variety of lipids did not interfere with the formation of color by standard sulfatides. Sulfur-containing amino acids, sugar sulfates, and sulfated polymers neither produced color nor interfered in its formation by sulfatide. Chloride and nitrate ions produced relatively little color.

The method is much more sensitive (lower limit, about 0.002 µmole) than most methods currently employed for the analysis of sulfatides and has a high degree of precision. It is applicable to the analysis of sulfolipids in tissue extracts and gives values similar to those obtained by previously published procedures. The reliability of the method is increased when it is applied to partially purified lipid solutions. Sulfatides added to extracts of rabbit brain tissue were quantitatively accounted for by the assay. In addition, the method can be applied to samples obtained directly from thin-layer plates.

Supplementary key words colorimetric analysis • azure A • cerebroside sulfate • brain • sulfolipid • sulfatide • cholesteryl sulfate • fatty alcohol sulfates • gray matter • white matter • rabbit

Submitted on October 6, 1967
Accepted on January 11, 1968

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