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The neutral lipids of the erythrocytes were investigated in several common mammals: cow, dog, goat, horse, pig, rabbit, rat, and sheep. Cholesterol content was determined by gas-liquid, thin-layer, and column chromatography, the last in conjunction with the IR spectrophotometry. The three methods yielded similar results. In every species investigated, cholesterol was the major neutral lipid; cholesteryl esters, triglycerides, and free fatty acids were detected only in trace amounts. It is concluded that these substances may have been contaminants from plasma lipoproteins or leukocytes rather than true constituents of the erythrocyte. In the erythrocytes of all species, cholesterol content was close to 30% of the total lipids extracted from the cells, and the molar ratio of cholesterol to phospholipid was approximately one. The significance of the data is discussed in relation to current concepts of the structure of the cell membrane involving cholesterol-polar lipid complexes.