Enhanced Detection of Curcumol in Biological Samples via Acetonitrile‐Anhydrous Sodium Sulfate Salt‐Assisted Liquid‐Liquid Microextraction Coupled With High‐Performance Liquid Chromatography

ABSTRACT Curcumol, a significant bioactive molecule, requires accurate detection methods due to its extensive pharmacological properties. This study develops high‐performance liquid chromatography (HPLC) methods to detect curcumol and constructs sample pretreatment techniques using salt‐assisted liquid‐liquid microextraction (SALLME). The optimization process focuses on conditions for acetonitrile (ACN) anhydrous sodium sulfate‐assisted liquid‐liquid microextraction. By adding excess anhydrous sodium sulfate to an ACN aqueous solution, ACN precipitates, forming a saturated saline solution and resulting in a two‐phase system. Curcumol, being highly soluble in ACN, separates into and enriches within the ACN phase, achieving effective matrix separation. SALLME demonstrated an enrichment factor of up to 12.4 for curcumol. The extracted ACN is then injected into an HPLC instrument for quantitative detection and analysis. The method exhibited a good linear range (0.5–10.0 µg/mL) and a low detection limit (0.05 µg/mL). The results indicate that curcumol can be effectively separated from complex matrices using this technique. By optimizing salt concentration and temperature, the HPLC method for determining curcumol in biological samples was successfully established, achieving recoveries between 86.2% and 137.6% in spiked urine and blood samples. Furthermore, this method offers rapid, efficient, and environmentally friendly sample preparation, promising significant applications in pharmacological and clinical settings.