Paper chromatography of lipides

Schlenk, Hermann; Gellerman, Joanne L.; Tillotson, Jerry Ann; Mangold, Helmut K.

doi:10.1007/bf02637887

Cited by 90 publications

(26 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Over 85 per cent of the label was in the triglyceride fraction, about 10 per cent in the diglyceride fraction, and 1 to 2 per cent in both monoglyceride and phospholipid. In view of the very small proportion of counts in the lower glyceride fractions, their identity was also confirmed by chromatography on silicone impregnated paper (14). Some of the monoglyceride was found to come from hydrolysis of higher glycerides on the IRA-400 column.…”

Section: Intestinal-lipid Metabolism and Bile Saltsmentioning

confidence: 84%

Studies on Lipid Metabolism in the Small Intestine With Observations on the Role of Bile Salts*†

Dawson¹,

Isselbacher²

1960

J. Clin. Invest.

269

View full text Add to dashboard Cite

It is now well established that the esterification of long chain fatty acids is an important step in their transport across the small intestinal mucosa before they appear in the lymph in chylomicrons. Evidence for this process has been well reviewed recently (1, 2). The main ester formed by the mucosa is triglyceride but a small proportion of the fatty acids is incorporated into phospholipids and cholesterol esters. We have recently reported some in vitro studies on the incorporation of long chain fatty acids into glycerides using homogenates of rat and human small intestine mucosa and have delineated some of the cofactor requirements involved (3). This work has now been extended to the intact mucosa of the rat. Observations are presented on factors influencing the uptake and esterification of palmitate-1-C4 and the incorporation of label into lipid from C14-glucose. In addition, evidence is presented for the concept that conjugated bile salts directly stimulate glyceride metabolism in the intestinal mucosa. MATERIALS AND METHODSPalmitic acid-1-C14 and uniformly labeled glucose-C4 were obtained from the Volk Radiochemical Company, Chicago, Illinois. The C14-labeled palmitate was purified and made up into a solution as previously described (2 purity tested by melting point determinations and paper chromatography (5). Hydrolysis of the conjugated bile salts, glycocholate and taurodesoxycholate was carried out by dissolving them in 2 N NaOH and then heating in sealed Pyrex tubes at 140°C for 3 hours. The resulting solution was diluted with water, acidified, and the precipitated free bile salt filtered, washed and dried. In earlier experiments commercial sodium taurocholate (Pfanstiehl Laboratory, Waukegan, Ill.) was used. This contained a faint trace of glycocholate which could only be demonstrated by chromatography when there was heavy overloading of Whatman no. 3 filter paper. No unconjugated bile salts were present but there were some contaminating pigments. The commercial taurocholate gave substantially the same results as the synthetic material which was used in later experiments.Female albino rats (Charles River Laboratories, Boston, Mass.) weighing 150 to 250 g were fasted overnight. They were killed by a blow on the head and the small intestine washed out at room temperature with oxygenated Krebs-Ringer phosphate buffer, pH 7.4, modified to contain half the usual concentration of calcium. This buffer was used in all the experiments described. The small intestine was everted on a glass rod following the technique of Wilson and Wiseman (6) and then cut across so as to form small cylindrical segments or intestinal rings. These measured approximately oneeighth inch (lengthwise) and weighed between 80 to 150 mg (wet weight). Since one of the problems in using slices or sacs of small intestine in metabolic studies is the variation in activity along this organ, some preliminary experiments were performed to determine the distribution of esterifying activity along the intestine. It was found that under optimal cond...

show abstract

Section: Intestinal-lipid Metabolism and Bile Saltsmentioning

confidence: 84%

Studies on Lipid Metabolism in the Small Intestine With Observations on the Role of Bile Salts*†

Dawson¹,

Isselbacher²

1960

J. Clin. Invest.

269

View full text Add to dashboard Cite

show abstract

“…These bands may have been due to polymerized fatty acids. It has been demonstrated that in reversed phase paper chromatography the polymerized substances stay at the starting point of the chromatograms (47). In countercurrent distribution, the polymerized substances would be expected to favor the less polar moving phase.…”

Section: Resultsmentioning

confidence: 99%

The Fatty Acids of Total Lipids and Cholesterol Esters from Normal Plasma and Atheromatous Plaques1

Tuna¹,

Reckers²,

Frantz³

1958

J. Clin. Invest.

View full text Add to dashboard Cite

In order to elucidate the mechanism of atheroma formation in arteriosclerosis, the lipid fractions of plasma and atheromatous plaques have been extensively studied in the past 45 years. It was first noted that there is a high content of cholesterol, phospholipid and total extractable lipids in atheromatous aortas (1-4). Then came a period when studies dealt primarily with the changing ratios between total, free and esterified cholesterol and phospholipids in normal plasma and atheromatous plaques (5-8). Considerable work has been done in the past decade to correlate the changes in blood cholesterol, phospholipids and lipoproteins with the incidence of arteriosclerosis (9-15). Comparatively little work has been done, however, on the isolation and identification of fatty acids associated with different lipid fractions of plasma and atheromatous plaques, the lack of adequate methods for fatty acid analysis being the most likely explanation. Schoenheimer (3) isolated the cholesterol esters of stearic, oleic and palmitic acids from the adventitia-free atheromatous aorta. In addition to these acids, McArthur (16) isolated linoleic and arachidonic acids from aortic plaques. As far as the fatty acids of the blood are concerned, the main sources of information are alkali isomerization studies (17-19) and studies done on the blood of large mammals (20). Recently, more information based on paper (21), column (22) and gas-liquid (23) chromatographic analysis of fatty acids of human blood has been reported. However, no comparative studies are available on material from human beings, using the same analytical methods to determine the fatty acids in blood and atheromatous plaques. There have been isolated reports suggesting that the blood of popu-

show abstract

“…methanol. The column was initially standardized with mono-, di-, and triolein and the eluates checked by chromatography on silicone impregnated paper (26).…”

Section: Methodsmentioning

confidence: 99%