The effect of sulfur limitation on lipid accumulation of Trichosporon fermentans CICC 1368 were investigated. Firstly， lipid content in microbial cells at the end of fermentation was determined using low field nuclear magnetic resonance （LF-NMR）. Then the cell biomass， lipid concentration and fatty acids compositions were assessed. Finally， the lipid accumulation was monitored at the optimum C/S molar ratio. Results showed that LF-NMR can be used as a rapid and effective method to analyze lipid content in microbial cells. Sulfur could effectively control the accumulation of lipids. Lipid content and yield increased first and then decreased as the C/S molar ratio raised， and this indicated that limitation of sulfur should be controlled in an appropriate range. The proper limitation of sulfur would promote the accumulation of the total unsaturated fatty acids， and oleic acid content was up to a maximum of 54.9%. The biomass， lipid concentration， lipid content at a C/S molar ratio of 3310 were 21.1 g/L， 9.0 g/L， 42.3%， respectively.