The variation of the berberine content in roots and stem bark of Berberis asiatica with altitude and edaphic conditions in the western Himalaya was estimated by HPLC. The comparative assessment revealed a significantly higher berberine content in roots than in stem barks. Moreover, the berberine content varied significantly with altitude and edaphic conditions both in root and stem bark samples. The populations growing at low altitude contained significantly more berberine than the ones growing at high altitude. Also the moisture and potassium (K) percentage of the soil significantly influenced the berberine content.