Purification of Curcumin, Demethoxycurcumin, and Bisdemethoxycurcumin by High-Speed Countercurrent Chromatography

• Published in: Journal of agricultural and food chemistry Vol. 56; no. 20; pp. 9328 - 9336
• Main Authors: Inoue, Koichi, Nomura, Chihiro, Ito, Sumio, Nagatsu, Akito, Hino, Tomoaki, Oka, Hisao
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 22-10-2008
• Subjects: Analytical Methods; antioxidant activity; Biological and medical sciences; bisdemethoxycurcumin; chemical composition; Chromatography, High Pressure Liquid; countercurrent chromatography; Countercurrent Distribution - methods; curcumin; Curcumin - analogs & derivatives; Curcumin - chemistry; Curcumin - isolation & purification; demethoxycurcumin; food analysis; food composition; Food industries; Fundamental and applied biological sciences. Psychology; high-speed countercurrent chromatography; liquid chromatography−tandem mass spectrometry; purification; turmeric; Turmeric powder; Turmeric powder; bisdemethoxycurcumin; high-speed countercurrent chromatography; liquid chromatography−tandem mass spectrometry; curcumin; demethoxycurcumin; Countercurrent chromatography; Purification; Liquid chromatography; Powder; Turmeric; High speed; liquid chromatography-tandem mass spectrometry; Curcumin; Mass spectrometry
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/jf801815n

Curcuminoids are substances of great interest because of their important pharmacological activities, particularly anti-inflammatory, anticarcinogenic, and anti-Alzheimer’s activities. In this study, we report the first procedure and effect of processing for the high, efficient, and useful purification of curcumin, demethoxycurcumin, and bisdemethoxycurcumin from turmeric powder. Purification involves high-speed countercurrent chromatographic (HSCCC) separation of these curcuminoids using a simple two-phase solvent system composed of n-hexane/chloroform/methanol/water (5/10/7.5/2.5, v/v). The HSCCC-fractionated effluent peaks indicated that the peak resolutions were 1.7 between curcumin and demethoxycurcumin and 2.1 between demethoxycurcumin and bisdemethoxycurcumin for 25 mg of loaded turmeric powder. These purified substances were analyzed by liquid chromatography−tandem mass spectrometry with scan and daughter scan negative modes, and the wide absorbance from 200 to 500 nm was monitored by photodiode array detection. The separation yielded 1.1 mg of curcumin, 0.6 mg of demethoxycurcumin, and 0.9 mg of bisdemethoxycurcumin (>98% purity). Moreover, the antioxidant effect of curcuminoids was measured by a 1,1-diphenyl-2-picrylhydrazil assay. The order of antioxidant activity was purified curcumin > purified demethoxycurcumin > purified bisdemethoxycurcumin > turmeric powder. Curcumin, demethoxycurcumin, and bisdemethoxycurcumin can be used for various evaluations of their pharmacological activities.