Isolation, identification and quantification of brazilein content in sappan wood (Caesalpinia sappan) traditional drink using LC HRMS and HPLC method

For decades, the demand for attractive colors in foods and beverages has been steadily increasing. Brazilein, a red pigment extracted from the heartwood of Caesalpinia sappan L. (CS), is commonly used in various traditional drinks, including the Indonesian herbal beverage wedang uwuh. However, brazilein's red color is notably unstable due to its sensitivity to heat, light, oxygen, and pH fluctuations, making it challenging to analyze. This study aimed to determine the brazilein content in both CS heartwood (a single-source material) and wedang uwuh (a mixed-source product). Before this, the samples underwent blanching and drying at various temperatures before extraction. The brazilein was extracted using the maceration method employing 96 ethanol for 72 hours. The resulting extracts were analyzed using High-Performance Liquid Chromatography (HPLC). The result showed that the measurement was validated and exhibited a Correlation Coefficient (R2) of 0.9997, Limit of Detection (LOD) of 0.0095 (µg/mL), and Limit of Quantification (LOQ) of 0.0287 (µg/mL). The brazilein concentrations varied with drying temperature, ranging from 2.5383±0.01 to 3.0155±0.01 (w/w) in CS and from 1.6335±0.01 to 3.0168±0.01 (w/w) in wedang uwuh. Notably, the lower brazilein content in wedang uwuh reflects its complex composition and the additional processing steps it undergoes, which likely contribute to pigment degradation. © 2025, Instituto de Tecnologia de Alimentos - ITAL. All rights reserved.