A standardized extract from Sesbania agati leaves (SESAL) was developed by Zenherb labs Pvt. Ltd., Mumbai, and is one of its kind plant-based biotin supplements. The current work aimed at characterization of SESAL using chromatographic techniques (like HPLC & HPTLC) and DNA authentication. Additionally, an in-silico approach (docking) was also adopted to predict the molecular interactions and probable mechanisms involved in potential role of SESAL in hair care products range. Drug likeliness was assessed on the basis of ADMET properties and Lipinski’s Rule of 5. DNA fingerprinting followed by HPTLC confirmed the identification and authentication. The biotin content in SesZenBio was estimated to be 0.5% by validated HPLC method. Mechanistically, biotin along with 2 other cofactors in SesZenBioTM is predicted to stimulate the hair follicle growth and differentiation, thus improving hair health. SesZenBioTM might hold 2x potential as against synthetic biotin in hair care applications.