Separation and purification of tricin from an antioxidant product derived from bamboo leaves

Tricin (5,7,4'-trihydroxy-3',5'-dimethoxyflavone) occurs in its glycosidic form in rice bran and other grass species such as wheat, barley, and maize. Tricin is considered sufficiently safe for clinical development as a cancer chemopreventive agent, therefore it can be used for cancer prevention. This study established a new method for the preparation of tricin from bamboo leaves as an alternative to traditional methods such as chemical synthesis via the Baker-Venkata-Raman reaction between acetylsyringic acid and phloroacetophenone. Tricin was prepared from an antioxidant product that was derived from bamboo leaves (AOB) by extraction with aqueous ethanol. A concentrated solution of this product was obtained and then processed by polystyrene (AB-8) resin column chromatography and preparative high-performance liquid chromatography (HPLC) with 30% (v/v) acetonitrile in 1% (v/v) acetic acid as the mobile phase. The collected tricin-rich fraction was further sequentially purified by dialysis membrane separation and drowning-out crystallization methods. The purity was assessed by analytical HPLC with 25% (v/v) acetonitrile in 1% (v/v) acetic acid as the mobile phase, and the chemical confirmation was evaluated by infrared, mass, nuclear magnetic resonance, and ultraviolet spectroscopies. Tricin (3.09 g) was prepared from 174 g of a crude column chromatography fraction obtained from 5 L of AOB concentrated solution. The present method is appropriate for preparing quantities of pure tricin, which can be used for the quantification of tricin in various plant herbs and further for pharmaceutical/biomedical applications and evaluation.