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ARS Home » Northeast Area » Boston, Massachusetts » Jean Mayer Human Nutrition Research Center On Aging » Research » Publications at this Location » Publication #203631
Title: Effects of Mobile Phase Ratios on the Separation of Plant Carotenoids by HPLC with a C30 Column

Author
item LI, LEI - CHINESE CDC
item QIN, JIAN - HNRCA AT TUFTS
item YIN, SHUAN - CHINESE CDC
item Tang, Guang-Wen

Submitted to: Chromatographia
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/2/2006
Publication Date: 11/29/2006
Citation: Li, L., Qin, J., Yin, S., Tang, G. 2006. Effects of Mobile Phase Ratios on the Separation of Plant Carotenoids by HPLC with a C30 Column. Chromatographia. 65(1/2):91-94.

Interpretive Summary: Some unidentified components in plant products often affect an accurate quantitative determination of lutein and zeaxanthin. A high performance liquid chromatography (HPLC) system equipped with a Bischoff C30 column was used to separate lutein, zeaxanthin and other unidentified components in plant products. Mobile phase A containing methanol, MTBE and water with a ratio of 60:33:7 by vol. (1.5% ammonium acetate, NH4Ac), combined with mobile phase B with a ratio of 9:90:2 by vol. (1.0% NH4Ac) is optimal for the separation. This method was successfully applied to the quantitative determination of lutein and zeaxanthin in extracts of plant products, such as chlorella, spirulina, celery and mallow.

Technical Abstract: Plant products are dietary sources of lutein and zeaxanthin. Lutein and zeaxanthin have been implicated in the protection of age related macular degeneration (AMD) and in cardiovascular diseases. However, xanthophylls and unidentified components (lambda-max = 423 nm and lambda-max = 468 nm) in plant products are often not separated well, and affect an accurate quantitative determination of lutein and zeaxanthin. A high performance liquid chromatography (HPLC) system equipped with a Bischoff C30 column and a mobile phase of methanol, methyl-tert-butyl ether (MTBE) and water was used to separate lutein, zeaxanthin and other unidentified components in plant products. Mobile phase A containing methanol, MTBE and water with a ratio of 60:33:7 by vol. (1.5% ammonium acetate, NH4Ac), combined with mobile phase B with a ratio of 9:90:2 by vol. (1.0% NH4Ac) is optimal for the separation. This method was successfully applied to the quantitative determination of lutein and zeaxanthin in extracts of plant products, such as chlorella, spirulina, celery and mallow.