A total of 80 green coffee bean (Coffea arabica L.) samples were five potential coffee-producing areas of East Gojjam, West Gojjam, Awi, Bahir Dar special zone and South Gondar zones of Amhara region. Coffee biochemical in this region has not yet been characterized and comprehensive profile of the constituents has not been reported. Therefore the main aim of this study was to provide comprehensive data on Trigonelline, Chlorogenic acid and Caffeine content in Amhara region’s coffee bean samples. The Caffeine, Trigonelline and Chlorogenic acids were simultaneously analyzed by HPLC-VWD. The results Coffees of the studied districts were rich in Caffeine, Trigonelline and Chlorogenic acids and it lies within the range of other reported data on Arabica coffee. Trigonelline ranged from 0.53 - 1.27g/100g, Chlorogenic acids ranged 3.29 - 7.73g/100g and Caffeine range 0.77 - 1.55g/100g. In the study area, Bahir Dar has lower Trigonelline, Chlorogenic acids and Caffeine concentration compared to the others. Whereas Awi zone’s coffee had higher Caffeine and East Gojjam had a maximum concentration in Chlorogenic acids and Trigonelline.