The phospholipids in human blood fractions Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/zp38wg99s

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  • Although there is information available on the distribution of phospholipids in human serum and red cells, very limited data have been reported for white cells and platelets. To the knowledge of the author, no data have been reported on the distribution of phospholipids among the four blood fractions, of individual subjects. Due to the limited amounts of white cells and platelets present in blood, micromethods are essential for the analyses of these fractions in individual subjects. Although phospholipids have been separated from larger samples by others, their methods were not appropriate for micro amounts. Therefore, procedures were developed in this study which made it possible to isolate and to quantitate the individual components of samples of total phospholipid ranging from 20 to 40 μg. The distributions of phospholipids were determined in serum, red cells, white cells and platelets isolated from the venous blood of four men and four women. The blood fractions were isolated and lipid was extracted by methods previously developed in this laboratory. Total phospholipids were isolated by preparative thin-layer chromatography. Individual phospholipid components were separated and quantitated by the micromethod developed in this investigation. Components were eluted from microchromatoplates and quantitated by analysis of their phosphorus content. This method effectively separated phospholipid samples into lysophosphatidylcholine (LPhC), sphingomyelin (Sph), phosphatidylcholine + phosphatidylserine (PhC + PhS) and phosphatidylethanolamine (PhE). Although all of the fractions contained higher proportional amounts of PhC + PhS than any other component, there were characteristic distributions in each of the blood fractions. Serum was characterized by high PhC + PhS and virtually no PhE. Red cells contained lower amounts of PhC + PhS and more Sph and PhE than any other blood fraction. The distributions of phospholipids in white cells and platelets were similar and resembled the general pattern found in red cells more than that of serum. The marked differences in the distribution of phospholipids among the blood fractions emphasize the importance of concurrent analyses of all blood fractions in studies of human phospholipid metabolism.
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