Questioned document examination is a term for a forensic science discipline pertaining to documents that are potentially disputed scientific processes and methods. Handwriting is a person’s writing created with writing instruments such as pen, pencil etc. on the writing surface. It is a visual speech of the individual, which is non-spoken but is written with the help of writing instrument. The study of handwriting, especially when employed as a mean of analyzing writing characters is known as Handwriting Analysis. The present work focuses on the comparative analysis of handwriting of students among two different medium (i.e. Hindi medium and English medium). The handwriting of every person individually acquired as the nervous system through hand. During the writing process various writing characteristics of handwriting are exhibited, from which identification of writer is ascertained. These characteristics features present in class and individual characteristics in the handwriting of two different medium’s students. In the present study, the writing sample was collected randomly by hundred individuals from two different mediums to find out the unique characteristics for differentiation of Hindi and English medium handwriting. The outcome of present study may give important information about various characteristics features in handwriting of Hindi medium and English medium students which will help the handwriting expert to establish whether the questioned writing has been from Hindi medium writer or English medium writer and this study also play a significant role in juvenile delinquency.