Mung　bean　sprouting　significantly　modifies　the　composition　of　free　amino　acids　(AAs),　making　studying　its　dynamic　changes　crucial　for　food　nutrition,　food　processing,　and　plant　metabolism.　Utilizing　derivatization　with　chiral　α-methoxy-α-trifluoromethylphenylacetyl　chloride,　ultra-performance　liquid　chromatography-tandem　mass　spectrometry　(UPLC-MS)　and　matrix-assisted　laser　desorption/ionization-trapped　ion　mobility　spectrometry-mass　spectrometry　(MALDI-TIMS-MS)　methods　were　developed　for　enantiomeric　AAs　analysis.　Mechanism　study　revealed　that　the　structural　differences　of　the　derivatized　AA　enantiomers　drive　their　separation.　UPLC-MS　with　a　C18　column　resolved　twelve　nonpolar　AAs　within　30　min,　achieving　detection　limits　of　1.4–4.1　ng/L.　MALDI-TIMS-MS　allowed　rapid　separation　and　spatial　mapping　of　the　same　set　of　AAs　under　fixed　conditions　with　little　sample　preparation.　The　derivatization　strategy　provides　a　novel　approach　for　chiral　analysis　and　chiral　metabolomics.　©　2025　Elsevier　B.V.