Rosemary (Rosmarinus officinalis) belongs to the Lamiaceae family, and is native to the Mediterranean and one of the most important medicinal herbs containing antioxidants in its leaves. One of the most important antioxidants is rosmarinic acid (RA). The aim of this study was to test the concentration of (RA) and chlorophyll content in leaves and callus of five successive subcultures of five different genotypes of rosemary. They were: 1) `Majorca'; 2) Rosmarinusofficinalis; 3) `Pine Scented'; 4) `Madeline Hill', and 5) APR. It was found that the highest concentration of RA in leaves was in `Pine Scented', while the lowest concentration was for APR and `Madeline Hill'. However, in the callus the highest RA concentration was for Rosmarinusofficinalis in the second subculture and `Madeline Hill' in the third subculture, while the lowest RA concentration was for `Majorca', `Pine Scented', and APR. The RA concentration in callus declined after the second and the third subculture for Rosmarinusofficinalis and `Madeline Hill', respectively. We concluded that it is preferred to use `Pine Scented' for RA extraction from the leaves while for RA extraction from callus it is better to use Rosmarinusofficinalis in the second subculture or `Madeline Hill' in the third subculture.