Solid phase fractionation techniques for segregation of red cabbage anthocyanins with different colorimetric and stability properties

Abstract

Red cabbage anthocyanin extract contains cyanidin derivatives as various non-, mono-, and di-acylated forms capable of expressing various colors from red to blue. The objective of this study was to develop a simple solid phase extraction (SPE) method to fractionate and segregate di-acylated anthocyanins which have greater stability and more desirable blue colorimetric properties. Two different sorbents with different separation chemistries: SCX (Strong Cation Exchange) and C18 sorbents were evaluated. Red cabbage anthocyanins were fractionated and eluted from the SCX cartridge using combinations of different buffers (pH 6-8) and MeOH (20-70%) or eluted from the C18 cartridge using different percentages of acidified (0.01% HCl) MeOH (30-100%) or EtOH (10-100%). With SCX SPE, washing the loaded pigments with a mixture of buffer pH 6 + 20% MeOH followed by elution with buffer pH 8 + 70% MeOH proved most effective in segregating the di-acylated pigments; however, some anthocyanins were permanently retained by the sorbent. When using the C18 cartridge, washing bound pigments with 32% MeOH or 18.5% EtOH and eluting with 100% alcohol respectively resulted in high (> 93%) recovery of di-acylated pigments. The C18 cartridge with MeOH eluent was the most effective for isolating the target di-acylated red cabbage anthocyanins that produced desirable blue colors with increased stability.

Keywords: Fractionation; Hydrophobic interaction; Solid phase extraction; Stable blue color; Strong cation exchange; di-acylated anthocyanins.