A series of long chain N-acyl amino acid type anionic surfactants were prepared by treating fatty acid chlorides with three kinds of amino acids, that is, sodium N-acyl-sarcosinates, sodium N-acyl-N-methyl-β-alaninates and sodium N-acyl-N-methyl-taurates in an alkaline solution. All prepared biodegradable surfactants were purified by thin layer chromatography and column chromatography, and identified their structures by spectral analysis.