In this paper, an inexpensive, rapid and efficient three phase partitioning (TPP) technique was used to simultaneously purify aloe polysaccharide (APS) and protein using a single-step extraction. This TPP system is established by adding ammonium sulfate and t-butanol to the crude slurry of aloe powder. APS was extracted in the lower phase, and the protein was extracted in the middle phase. The extraction conditions optimized were mass concentration of ammonium sulfate and t-butanol, temperature and pH. APS was further purified using a dialysis membrane to remove the salt, and its purity was determined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze aloe protein. TPP is an attractive and potential technique for the purification of APS and protein from Aloe vera L. (C) 2015 Elsevier Ltd. All rights reserved.