Glutathione　(GSH),　homocysteine　(Hcy)　and　cysteine　(Cys)　play　important　roles　in　many　physiological　processes.　However,　due　to　their　structural　and　functional　similarities,　it　is　still　a　challenge　to　develop　a　probe　that　can　differentiate　between　GSH　and　Hcy/Cys　simultaneously.　In　this　work,　a　luminescent　probe　Ir-NBD　was　designed　and　synthesized,　which　emit　weakly　due　to　the　presence　of　photo　induced　electron　transfer　(PET)　interaction.　When　it　reacted　with　the　three　biothiols,　NBD　dissociated　and　luminescence　of　Ir-OH　was　enhanced　in　the　near-infrared　(NIR)　region　due　to　the　disappearance　of　the　PET　effect.　On　the　other　hand,　the　products　obtained　from　the　reaction　of　GSH　with　NBD　were　hardly　luminescent,　but　the　products　from　the　reaction　of　Hcy/Cys　with　NBD　could　undergo　an　intramolecular　rearrangement,　resulting　in　an　enhanced　luminescence　of　the　solution　in　the　visible　region.　Ir-NBD　enabled　highly　selective　and　sensitive　detection　of　GSH　and　Cys/Hcy　in　a　relatively　short　time　(15　min).　The　two　luminescent　colors　were　clearly　differentiated　without　spectral　interference　and　the　detection　limit　reached　1.32　μM　(GSH),　0.42　μM　(Hcy)　and　0.51　μM　(Cys),　respectively.　Ir-NBD　also　had　low　cytotoxicity,　it　realized　the　simultaneous　detection　of　GSH　and　Hcy/Cys　by　dual-channel　luminescence,　and　also　provided　ideas　for　the　design　of　multifunctional　luminescent　probes.　©　2023　Elsevier　B.V.