The effect of n-butanol on Triton X-114 phase partitioning

n-Butanol interferes with the fractionation of amphiphilic and hydrophilic molecules during the Triton X-114 phase separation procedure. The indicators oil red (hydrophobic) and p-nitrophenol (hydrophilic) were useful for predicting the effectiveness of the Triton X-114 partition method. For n-butanol extracts containing oil red, 5'-nucleotidase, or alkaline phosphatase, the hydrophobic molecules and Triton X-114 were retained in the aqueous phase during incubations at 30 degrees C. The n-butanol interference was concentration-dependent and was reduced by lowering the final n-butanol concentration of the sample to 1.5% (v/v) or less. The results demonstrate how buffer-diluted n-butanol extracts of 5'-nucleotidase and alkaline phosphatase can be successfully employed for subsequent Triton X-114 fractionation of the enzymes.