The present work was aimed at formulation development, evaluation and MTT assay of Herbal Vitamin C Powder. The formulation was prepared by mixing Acerola Extract, Rosehip Extract, Moringa Extract, Amla Powder, Bilberry Extract, Glycine, L- lysine, L- proline, Elaichi Powder, Rock salt, Spirulina Powder and excipients such as Lemon flavor. Initially, the preformulation evaluation of each separate ingredient was done by evaluating its Organoleptic characters, Bulk density, Tap density, Hausner Ratio, Moisture Content, pH, and Solid-state stability study. The formulation was prepared after the pre-formulation study was completed and then evaluated for powder characters and MTT assay. Powder evaluation was performed using various physiochemical and microbiological parameters and the cytotoxicity of the formulation was performed by an MTT assay on Human embryonic kidney cells (HEK). From the above study, we can conclude that the stable polyherbal formulation of Vitamin C was prepared and evaluated and did not exhibit cytotoxicity at the daily dose limit.