Tyrosinase-based TLC Autography for anti-melanogenic drug screening 2025-05-01

Tyrosinase-based TLC (thin layer chromatography) was developed for screening of anti-melanogenicdrugs. In particular, this technique enables researchers to identify melanogenic inhibitor(s) in testedmixtures with the naked eye. In comparison with traditional colorimetric screening assays for tyrosinaseinhibitor(s), not only is tyrosinase-based TLC a more cost-efective option (nearly one-tenth theenzyme cost of colorimetric methods) but also is a more sensitive detection approach for kojic acid(KA), a standard anti-melanogenic drug. The detection limit of tyrosinase-based TLC and colorimetrictyrosinase assay for KA was 0.0125 and 1.25μg, respectively, demonstrating that the former was 100-fold more sensitive than the latter to determine the tyrosinase inhibitory rate of KA. Furthermore, theresults of this method have demonstrated excellent precision by Gage Repeatability and Reproducibility(Gage R&R), with the variation of total Gage R&R being 28.24%. To verify the applicability oftyrosinase-based TLC, this platform was employed to screen melanogenic inhibitor(s) from Ganodermaformosanum extracts and two of all fractions (GFE-EA F4, F5) obtained showed depigmenting activity. Itis noteworthy that these two fractions also exerted anti-melanogenesis activity on zebrafsh, thereforeverifying the credibility of tyrosinase-based TLC. In sum, this technique provides new insight into thediscovery of novel melanogenic inhibitor(s).