Abstract
Preparative silicic acid-Hyflo Supercel column chromatography of crude lipid extracts of porcine intestine gave two glycolipid fractions. The larger fraction was separated into hematosides, porcine intestinal glycolipids I & II (PIGL-I & PIGL-II) and sphingomyelin by rechromatography on a silicic acid-Hyflo Supercel column and a DEAE-cellulose column. Final purification of these glycolipids was achieved by thinlayer chromatography. The amounts of PIGL-I & PIGL-II were found to be more than half of the total sphingoglycolipids containing hexosamine or sialic acid, although other glycolipids of this type, hematosides and globoside I, were also isolated in appreciable amounts. From the smaller fraction, cerebrosides (CMH), ceramide dihexosides (CDH), ceramide trihexoside (CTH), cerebroside sulfate (CMHS) and globoside I were isolated by means of rechromatography with silicic acid-Hyflo Supercel, and subsequently by column chromatography with DEAE-cellulose or Florisil. CDH, CTH and globoside I were finally separated by thin-layer chromatography. These ceramide hexosides also were found to be major glycolipids in porcine intestine. CMHS was however obtained in a small amount. These glycolipids were characterized further by qualitative and quantitative analyses as follows: ceramide glucoside & ceramide galactoside (CMH); 3 3 Following abbreviations are used in this paper: CM, a mixture of chloroform and methanol; 95 CM, 80 CM, 75 CM, etc., the numbers show the volume percentages of chloroform in the mixtures of chloroform and methanol; CMH, cerebroside (ceramide monohexoside); CDH, ceramid dihexoside; CTH, ceramide trihexoside; CMHS, cerebroside sulfate (sulfatide); PIGL, porcine intestinal glycolipid. ceramide galactoside sulfate (CMHS); ceramide lactoside & ceramide digalactoside (CDH); digalactosylglucosyl-ceramide (CTH); N-acetylgalactosaminyldigalactosylglucosyl-ceramide (globosidel); N-acetyl- and N-glycolyl-neuraminylgalactosylglucosylce-ramides (hematosides); glycolipid containing glucose, galactose, N-acetylhexosamine (glucosamine: galactosamine = 1:1), fucose, and ceramide in the molar ratio of approximately 1:2:1:1:1 (PIGL-I); glycolipid consisting of glucose, galactose, N-acetylglu-cosamine, fucose and ceramide in the molar ratio of about 1:2:2:1:1 (PIGL-II). The glycolipids showed large variation in fatty acid composition. Fatty acid compositions of PIGL-I and PIGL-II resembled each other, showing a similarity to that of CDH, but differed remarkably from those of globoside I and hematosides. On the basis of column and thin-layer chromatography and of analytical data, PIGL-I and PIGL-II were shown to be novel glycolipids. From the present observations and comparison with previous reports, the glycolipid composition of porcine intestine is species and organ specific.
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