A procedure for the screening and quantification of 18 glucocorticoids, i.e. hydrocortisone sodium succinate (HSS), prednisolone (PDL), prednisone (PDS), hydrocortisone (HCS), methylprednisolone (MPS), betamethasone (BTM), dexamethasone (DXM), triamcinolone acetonide (TA), prednisolone acetate (PLA), hydrocortisone acetate (HA), fludrocortisone acetate (FA), prednisone acetate (PA), cortisone acetate (CA), dexamethasone acetate (DA), hydrocortisone butyrate (HB), triamcinolone acetonide acetate (TAA), fluocinonide (FN) and halcinonide (HC) from herbal pharmaceuticals and health foods was established and fully validated. The samples were extracted by methanol and separated by HPLC. The retention times and ultraviolet spectra were used for the preliminary screening, and the suspected adulterants were then confirmed by liquid chromatography-quadrupole-time of flight mass spectrometry (LC-Q-TOF-MS/MS) and quantified by HPLC. The developed procedure was successfully applied to 14 herbal samples, and 316.3 µg g of DA and 13.6 µg mL of BTM were found in a tablet sample and a spray sample, respectively. To our best knowledge, this is the first report of the simultaneous screening and quantification of these 18 glucocorticoids from any matrix.