Identification of colored styles as one of the saffron adulteration with HPLC

High performance liquid chromatography (HPLC) with UV-visible detection was applied to identify adulterated saffron prepared by adding colored styles with the natural colorants extracted from saffron petals, safflower, madder and red beet to the pure saffron. The chromatograms of the methanol-water (50%, v/v) extracts of pure and adulterated saffron was obtained using a Nucleosil RP C18 column (5 m) and a linear gradient from 20 to 80% methanol in water in 60 min with the flow rate of 1 ml min-1 at temperature 30°C. Two-way analysis of variance (ANOVA) was then applied to the HPLC data of the extracts of pure and adulterated saffron at the assayed wavelengths, 520 (or 254), 402 (or 254), 260 and 535 (or 440) nm and the presence of colored styles with the colorants of saffron petals (>9.1%, w/w), colored styles with the colorants of safflower (>14.3%), colored styles with the colorants of madder (>9.1%) and colored styles with the colorants of red beet (>14.3%) in saffron were significantly identified, respectively.