Development and validation of high performance thin layer chromatographic method was carried out for the estimation of adulterants (sudan dyes) in turmeric and chili powdered spices. The separation of sudan dyes were obtained over aluminium baked HPTLC plates pre-coated with silica gel RP-18 F-254 as stationary phases with optimized triphasic mobile phase acetonitrile: acetone: ammonia (9.0:0.5:0.5; v/v/v) with 5 min saturation time. Validation was carried out by testing its specificity, linearity, accuracy, inter and intraday precision, repeatability, limit of detection and limit of quantification according to ICH guidelines. Recording of characteristic molecular ion peaks of sudan I, II, III and IV at 248, 278, 353 and 381, respectively highlights the use of TLC-MS interface confirming the adulteration of sudan dyes in some commercially available powders of spices.