Glyphosate is a frequently detected herbicide in forensic cases. In order to achieve an optimized and unified detection method of glyphosate and provide scientific evidence on glyphosate metabolic process in different organs of animals. Gas chromatography tandem mass spectrometry(GC-MS) was applied for analysis. And for derivatization, two methods including mixtures of trifluoroethanol(TFE)/trifluoroacetic anhydride(TFAA) and fluorenylmethyl chloroformate(FMOC-Cl) were utilized to increase volatility of the compound[1-3]. This study compared the existence of glyphosate and its metabolite aminomethyl phosphonic acid(AMPA) in four bio-samples from a goat forensic case[4-5]. In plasma, liver and lung, AMPA concentrations were detected more than glyphosate result from the deacetylation process(Fig.1)