Abstract: Astaxanthin have been extracted and purified from mutant isolate of Phaffia rhodozyma JH-82. Purified astaxanthin was identified by spectrophotometric, TLC and HPLC analysis and were compared with synthetic astaxanthin. Results of TLC analysis indicated that isolate of P. rhodozyma JH-82 were able to produce nine different carotenoids and high level of carotenoids was belong to astaxanthin. Results of this study for pure astaxanthin production indicated that mutant of JH-82 of P. rhodozyma (230 μg g-1 dried yeast) produced more astaxanthin than natural isolate JH-80 (140 μg g-1 dried yeast). The HPLC spectrum showed retention time 11 min for both purified and synthetic astaxanthin and solvent was CDCl3.