Abstract: In this study,the main signal peaks of~1 H nuclear magnetic resonance spectroscopy(~1H-NMR) in CDCl_3 extracts of butter were assigned by a few different NMR experimental methods.Then the fatty acid composition of butter were determined upon analysis of~1H-NMR spectra. Twenty-eight brands of commercially available butter were determined in the study. The results showed that the relative molar contents of linolenic acid,linoleic acid,butyric acid,oleic acid,conjugated linoleic acid( CLA) were 0.55% ~1.00%,0.59% ~3.20%,9.56% ~12.59%,13.18% ~21.01%,0.34% ~ 1.10%,and the contents of saturated fatty acids( SFA) and essential fatty acid( linolenic acid,linoleic acid,CLA) were 74.95% ~84.44% and 2.16% ~4.35%.In addition,1-pentene in CDCl_3 extracts of butter was identified by 1D-TOCSY,J-RES,~1H-~1H COSY spectra.The result suggest that this method can not only clarify the target but also non-target components in butter.