Purification of a Water Extract of Chinese Sweet Tea Plant (Rubus suavissimus S. Lee) by Alcohol Precipitation

The aqueous extraction process of the leaves of Rubus suavissimus often brings in a large amount of nonactive polysaccharides as part of the constituents. To purify this water extract for potential elevated bioactivity, an alcohol precipitation (AP) consisting of gradient regimens was applied, and its resultants were examined through colorimetric and HPLC analyses. AP was effective in partitioning the aqueous crude extract into a soluble supernatant and an insoluble precipitant, and its effect. varied significantly with alcohol regimens. Generally, the higher the alcohol concentration, the purer was the resultant extract. At its maximum, approximately 36% (w/w) of the crude extract, of which 23% was polysaccharides, was precipitated and removed, resulting in a purified extract consisting of over 20% bioactive marker compounds (gallic acid, ellagic acid, rutin, rubusoside, and steviol monoside). The removal of 11% polysaccharides from the crude water extract by using alcohol precipitation was complete at 70% alcohol regimen. Higher alcohol levels resulted in even purer extracts, possibly by removing some compounds of uncertain bioactivity. Alcohol precipitation is an effective way of removing polysaccharides from the water extract of the sweet tea plant and could be used as an initial simple purification tool for many water plant extracts that contain large amounts of polysaccharides.