To overcome the problem of relying on metal sol substrates in traditional thin-layer chromatography surface enhanced Raman spectroscopy, this study mixed AuNPs/MIL-101(Cr) composite with excellent SERS performance and chromatographic silica gel to form a homogenate, which was then spread on a glass slide to prepare MIL-101(Cr) modified thin layer chromatography plates with both intrinsic SER activity and good stability. Establish a MIL-101(Cr) modified thin layer chromatography-surface enhanced Raman spectroscopy method by combining a portable Raman spectrometer with a MIL-101(Cr) modified thin-layer chromatography plate. The innovation of this method lies in that before performing SERS detection on the sample, only 0.01mol/L hydrochloric acid solution needs to be added to wet the sample spots, and strong SERS signals can be collected without adding any SERS substrate. The results showed that the MIL-101 (Cr) modified thin layer chromatography-surface enhanced Raman spectroscopy method successfully detected dyed Croci Stigma with four common dyes through SERS in-situ detection, which can be used for simple, rapid, and sensitive identification of adulteration in traditional Chinese medicine staining. Compared with the traditional thin layer chromatography-surface enhanced Raman spectroscopy method, it can better meet the needs of rapid and sensitive on-site detection of traditional Chinese medicinal materials.