Behdan A, Snow L D
Biochemistry Laboratory, Louisiana Tech University, Ruston 71272, USA.
Bioseparation. 1997;7(1):57-60. doi: 10.1023/a:1007962520611.
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.
正丁醇在Triton X - 114相分离过程中会干扰两亲性分子和亲水性分子的分级分离。油红（疏水性）和对硝基苯酚（亲水性）指示剂可用于预测Triton X - 114分配法的有效性。对于含有油红、5'-核苷酸酶或碱性磷酸酶的正丁醇提取物，在30℃孵育期间，疏水性分子和Triton X - 114保留在水相中。正丁醇的干扰具有浓度依赖性，通过将样品的最终正丁醇浓度降低至1.5%（v/v）或更低可以减少这种干扰。结果表明，5'-核苷酸酶和碱性磷酸酶的缓冲液稀释正丁醇提取物可如何成功用于随后的酶的Triton X - 114分级分离。
