Triglycerides of cottonseed oil were separated by thin layer chromatography (TLC), and fractionated by high-performance liquid chromatography (HPLC) on the basis of partition numbers. From each fraction, it was fractionated again on the basis of acyl carbon numbers using gas liquid chromatography (GLC). The fatty acids of triglyceride for each partition number group were analyzed by GLC. From, these results, triglyceride constituents of cotton seed oil were estimated to be 37 kinds of triglycerides. The major triglycerides and their contents in cotton seed oil were as follows: 25.8%$(C_{16:0},\;C_{18:2},\;C_{18:2})$, 15.5%$(C_{18:2},\;C_{18:2},\;C_{18:2})$, 13.8%$(C_{16:0},\;C_{18:2},\;C_{16:0})$, 8.3%$(C_{18:2},\;C_{18:1},\;C_{18:2})$, 6.2%$(C_{18:2},\;C_{18:1},\;C_{18:1})$, 4.1%$(C_{18:1},\;C_{18:1},\;C_{14:0})$, 3.4%$(C_{16:0},\;C_{18:1},\;C_{16:0})$, 2.3%$(C_{18:1},\;C_{18:2},\;C_{16:0})$, 2.2%$(C_{18:1},\;C_{18:1},\;C_{18:1})$, 1.0%$(C_{14:0},\;C_{18:2},\;C_{18:1})$.