Kajal is a folklore herbal formulation that has been commonly used for eye care in all communities of Indian families since ancient times. It is also known as surma or kohl. There are several Muslim and Hindu countries like India, Egypt, Rome, China, Japan, South Asia, the Middle East, and Africa where the application of Kajal is a common practice and also used in many types of cosmetics formulation for beauty, prevention, and treatment of numerous eye diseases. In the Chitrakoot region, Kajal was also applied to newborn babies as well as small children and people for a multipurpose such as an antiseptic, and astringent, to improve eye health and traditions with religious and therapeutic significance. Tribal women prepare different types of Kajal with the help of herbal plants. One is known as a Belha Kajal in the local language. The present study was carried out with the aim of documentation of folklore practices, and preparation of herbal Kajal used by the villagers of the Chitrakoot region. For standardization of Herbal Kajal was performed based on various selected parameters like botanical identification, physicochemical parameters, heavy metal screening, HPTLC fingerprint profile, and antimicrobial activities as per the standard procedure of Ayurvedic Pharmacopeia of India.