Determination of natural colorants in plant extracts by high-performance liquid chromatography

The determination of the colouring compounds apigenin (1), lawsone (2), juglone (3) and indigotin (4) in plant extracts using HPLC–UV/Vis methods is reported. The methods were applied to the analysis of 1–4 in ethanolic and propylene glycolic extracts originating, respectively, from chamomile (Chamomilla recutita [L] Rauschert, Asteraceae), henna (Lawsonia inermis L., Lythraceae), walnut (Juglans regia L., Juglandaceae) and natural indigo (Indigofera sp., Fabaceae). In the case of the indigo extracts, an optimized acid hydrolysis was applied. HPLC separations were performed on a Hypersil ODS RP18 column using linear gradient elution programs. The detection limits for 1–4 were 0.11, 0.6, 0.10, 0.089 μg mL-1, respectively. The procedure did not involve any sample “clean-up” methods. The amounts of the colouring compounds ranged from 0.006 (3) to 0.13 mg mL-1 (4) in the ethanolic extracts and from 0.22 (2) to 1.44 mg mL-1 (4) in propylene glycolic extracts. The proposed HPLC methods are advantageous in terms of sample preparation and the selective separation of the compounds. The plant dye extracts are commonly used in hair colouring formulations. The results indicate that the methods developed may serve for the quantitative control of dying plants and cosmetic products.