Isolation and Characterization of the Major Natural Dyestuff Component of Brazilwood (Caesalpinia sappan L.)
Brazilwood has been traditionally used as food and beverage colorant in Indonesia. The ethanol extract of brazilwood (Caesalpinia sappan L.) was subjected to the investigation of major natural dyestuff component. The dried heartwood was subjected to an extraction with 50% ethanol (1:3 v/v) at 80°C for 30 min, followed by lyophillization to crystalize the extract. The extract components were separated by using Sephadex G-15 gel filtration chromatography eluted with 40% ethanol under UV-Vis detection. This step yielded 5 fractions, FI-FV, of which FII and FIII were the major fractions, comprising 25 and 70% by weight of the applied dried extract, respectively. Both fractions had different UV-Vis spectra as well as infrared spectra observed by FTIR. Fraction FIII which visually had a deep red color, had the strong absorption of carbonyl group in its IR spectra. This major dyestuff fraction was purified further by RP-HPLC with multiwavelength detector using a solvent mixture of 65% acetonitrile and 35% isopropanol as a mobile phase. The purified fraction was then characterized by FTIR and ESI-MS. The infrared and mass spectra confirmed that brazilein with a molecular weight of 284 was the major dyestuff component of the ethanol extract of brazilwood. This finding can be utilized for the production of brazilein standard for the laboratory use to analyze brazilein in foods and beverages.