Eight kinds of non-volatile organic acids including malic acid,fumaric acid,9-octadecenoic acid and so on in cut tobacco from fifteen different planting areas were determined by GC/MS.The methyl esterification conditions of non-volatile organic acids were optimized.The results showed that the optimal time,temperature of methyl esterification was 50 min and 70 ℃.The optimal volume proportion of boron trifluoride to methanol,RSD of the method,the recovery of non-volatille organic acid was 3∶7,3.34%-7.32%,and 79.50%-101.35%,respectively.