Isolation and purification of two tyrosinase inhibitors from camellia pollen by high-speed counter-current chromatography

Abstract

Bee pollen is a health food with a wide range of nutritional and therapeutic properties. However, the bioactive compounds of bee pollen have not been extensively revealed due to low efficacy in separation. High-speed counter-current chromatography (HSCCC) and solvent extraction were applied to separate tyrosinase inhibitors from camellia pollen in this study. The camellia pollen extracts prepared with petroleum ether, ethyl acetate, and n-BuOH have tyrosinase inhibitory activity. Acidic hydrolysis could promote the tyrosinase inhibitory activity of crude sample. Three fractions with tyrosinase inhibitory activity were separated from the hydrolysate by a one-step HSCCC procedure. Among the fractions, two chemicals were sufficiently purified and identified to be levulinic acid (LA) and 5-hydroxymethylfurfural (5-HMF). The recovery was 0.80 g kg-1 pollen for LA and 1.75 g kg-1 pollen for 5-HMF; and their purity was all over 98%. The study demonstrates that HSCCC method is powerful for preparative separation of tyrosinase inhibitors from camellia pollen.