For analysis of Xanthene dyes according to the developing solvent and adsorbent was applied to Thin-layer chromatography with silicagel and cellulose plate. Silicagel chromato-plate used were prepared under different condition of activation. Using eight developing solvent, the influence of the condition for activation upon the separation of Xanthene dyes was investigated. The results are shown in Table 3. Methyl ethyl ketone+Acetone+$H_2O$ (10:0.1:0.4) mixture and n-butanol+Ammonia water (4:1) mixture gave clear separation for Xanthene dyes, including Fluorescein, Erythrosine Rhodamin B, Eosine, Rose bengale, phloxine and Acid red those Rf values decrease in the described ordor. Methyl ethyl ketone+Acetone+$H_2O$ (10:0.1:0.4) was applied to two adsorbents which were purchased from different manufactures. The results of Chromatograms are obtained Figure 6.