Thin-layer chromatography method for identification of saffron (Crocus sativus Linn.) in Kumkumadi Tailam

Abstract Background Skin disorders are widespread, and there is increasing demand for reliable, standardized herbal therapeutics. Kumkumadi Tailam, a classical Ayurvedic polyherbal oil formulation, incorporates several botanicals with purported dermatological benefits, among which Saffron (Crocus sativus L.) is a principal constituent reputed for its antioxidant and complexion-enhancing properties. Despite its high market value largely due to Saffron, there is a lack of systematic identification method for Saffron in commercially available samples. Moreover, no marker based profiling, for critical marker of Saffron, is reported to ensure the efficacy of the product. Objective This study was designed to develop and apply a Thin Layer Chromatography based analytical method for the qualitative identification of Saffron in Kumkumadi Tailam marketed samples. Reliable identification of Saffron in Kumkumadi Tailam using High Performance Thin Layer Chromatography (HPTLC) facilitates quality assurance, enables detection of adulterants, strengthens consumer trust, and supports the development of regulatory standards. In addition, new simple and selective, validated HPLC methods are established for separation and identification of Safranal & Berberine in marketed formulations. Thus, chromatographic profiling will help to confirm both qualitative as well as quantitative identification/estimation of marker in finished formulation to ensure quality. Methods Simultaneous Chromatographic Profiling of Saffron and Kumkumadi Tailam was carried out using pre-coated silica gel 60F254 (20 cm X 10 cm, E.Merck). The TLC plates were developed in suitable mobile phase consisting of toluene, ethyl acetate and acetic acid in a 5 : 5 : 0.1 (v/v/v) ratio. Saffron and Kumkumadi Tailam were applied as 10 mm bands using CAMAG Linomat 5 applicator equipped with a 100 µL syringe. Detection was performed densitometrically and comparison was based on retention factor (Rf) values and reflectance spectral profiles. Quantitative assay of marker components (Safranal and Berberine) has been carried out using C18 Column by isocratic elution and detection at 308 nm and 350 nm respectively using UV detector. Results Distinct chromatographic bands corresponding to Saffron were observed in both the standard and Kumkumadi Tailam samples at identical Rf values of approximately 0.71. Reflectance spectra recorded at 254 nm for these corresponding bands exhibited high similarity, confirming the presence of Saffron-derived compounds in the tested formulation. The observed chromatographic patterns demonstrate reliable co-migration and spectral congruence between Saffron and the formulation extract. Amounts of Safranal and Berberine in Kumkumadi Tailam samples are 0.38 (%w/w) and 0.07 (%w/w) respectively by HPLC. Conclusion The findings confirm the presence of Saffron constituents in Kumkumadi Tailam based on matched Rf values of approximately 0.71 at 254 nm and spectral profiles using HPTLC. This method provides a straightforward and reproducible approach for Saffron detection in complex herbal matrices, facilitating enhanced quality control, adulteration screening, and establishment of regulatory standards for high-value Ayurvedic products. In addition, Marker Based Profiling ensure Quality of the Finished Formulations as well as devised as a critical tool for batch-to-batch consistency. Graphical Abstract