Isolation and purification of plastochromanol‐8 for HPLC quantitative determinations

Abstract

The study describes isolation and semi‐preparative HPLC scale purification of plastochromanol‐8 (PC‐8). The PC‐8 standard was obtained from flaxseed oil. The purity of the obtained PC‐8 standard was 93.06% determined by UV–Vis detection. The HPLC/MS n fragmentation pattern of PC‐8 side chain resulted in the successive release of eight isoprene fragments which confirmed the correct structure identification. The established PC‐8 molar absorption coefficient (3616.5 M −1 cm −1 ) will allow the precise quantitative analysis of this compound in real samples without the use of a standard. Usefulness of the obtained PC‐8 standard was verified by HPLC determination of PC‐8, tocopherols, and tocotrienols at the same time. The PC‐8 recovery was 107.08%, and limit of detection (LOD) and limit of quantitation (LOQ) were 0.75 and 2.27 µg/mL, respectively. Practical applications: There are no commercially available standards of plastochromanol‐8 (PC‐8), and therefore effective ways of isolation of this compound are needed. A semi‐preparative HPLC is one of the available options allowing obtaining pure PC‐8.