The rise in concerns regarding fluoride-related health risks has driven the development of non-fluoride toothpaste formulations. This study focused on the production of a non-fluoride toothpaste using natural abrasives and active phytochemical ingredients and assessed its biochemical properties, including antimicrobial activity, pH stability, and enzymatic interactions with oral substrates. The toothpaste was formulated using hydrated silica, xylitol, sorbitol, essential oils, and herbal extracts. Biochemical assessments revealed effective microbial inhibition against Streptococcus mutans and Lactobacillus species, a neutral to slightly basic pH (6.8–7.2), and minimal interference with salivary enzymes. The findings indicate that non-fluoride toothpaste can offer effective oral hygiene while minimizing fluoride-associated risks, suggesting potential for broader commercial application.