Abstract: The contents of reduced glutathione ( GSH) and protein thiols in prunus duleis, cashew nut, pistachio nut, macadamia nut were detected by pre-column derivatization HPLC method using 5, 5'-dithio-bis-2-nitrobenzoic acid ( DTNB) as derivatizing agent with concomitant cell rupture sample pretreatments.The results showed that detected nuts contained relatively abundant GSH and thiols in P-SH. The amounts detected were GSH ( 1.2303 ± 0.01126) μmol/g, P-SH ( 3.5414 ±0.0531) μmol/g for pistachio nut, GSH ( 0.3069 ± 0.0185) μmol/g, P-SH ( 1.9001 ± 0.0194) μmol/g for prenus duleis, GSH ( 1.5708 ± 0.0216) μmol/g, P-SH ( 0.5905 ± 0.0238) μmol/g for macadamia nut, GSH ( 0.3373 ± 0.0047) μmol/g, P-SH ( 2.2458 ± 0.0225) μmol/g for cashew nut respectively.The recoveries of the methods were 99.37% ¹01.45%, and detection limits were 0.1386 μmol/g for GSH and 0.2184 μmol/g for protein thiols.The results provided a useful data for the application of these nuts for the healthcare.