A 25 factorial design was used to evaluate the effect of methanol, ethanol, propanol, butanol and pentanol on the paper chromatographic separation of Arginine, Threonine, Leucine, Histidine, and Tryptophan. The most significant effects occur for the alcohols themselves with an increasing effect with an increase in carbon chain length. The major interaction terms involve the combinations of butanol and pentanol with the other alcohols or with themselves.