Reversed phase silica gel column was applied for purifying Cyclic adenosine phosphate (cAMP) with purity as index.The optimal purification parameters were determined with adsorption and desorption rate as indicators,and the results showed that the sample fluid velocity was 1mL/min,the sample concentration was 70μg/mL,pH 6,the flow velocity 3mL/min with 60％ methanol as eluent.Under these conditions,the purity of cAMP was increased from 0.098％ to 44.69％.