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Method for the Determination of β-Carotene in Supplements and Raw Materials by Reversed-Phase Liquid Chromatography: Single Laboratory Validation

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A single laboratory validation (SLV) study was conducted for a liquid chromatography (LC) method for the determination of total and all-trans-β-carotene in a variety of dietary supplements, including multivitamin tablets, softgels, capsules, and beadlet raw materials. Extraction variants were developed for the different types of supplements tested based upon the supplement type and level of β-carotene. Water dispersible formulations such as powders, emulsions, tablets, and capsules were enzymatically digested with protease and extracted with dichloromethane–ethanol. Oily suspensions were directly dissolved in dichloromethane–ethanol. After appropriate dilution or concentration, the extracts were chromatographed by using either a reversed-phase C18 column or, in products containing high amounts of α-carotene, a reversed-phase C30 column. The LC systems provided linear responses in the range of 0.1–50 μg β-carotene/mL. The main geometrical isomers of β-carotene (all-trans, 9-cis, 13-cis, and 15-cis) were well separated from each other and from other carotenoids such as α-carotene, cryptoxanthin, lutein, lycopene, and zeaxanthin. Duplicate determinations of total β-carotene performed by 2 technicians in 8 different test materials on 5 different days resulted in relative standard deviations of 1.2–4.4%. Recoveries determined for supplements and beadlet raw material spiked with β-carotene levels of 10 μg to 100 mg/test portion and 0.2–40%, respectively, ranged from 97.5 to 102.1%. On the basis of the accuracy, precision, and recovery results from the SLV study, the method is suggested for a collaborative study on the determination of total and all-trans-β-carotene in dietary supplements.

Document Type: Research Article

Affiliations: 1: DSM Nutritional Products Ltd, Research and Development, Analytical Research Center, PO Box 3255, CH-4002 Basel, Switzerland. 2: Caravan Products Co. Inc., 100 Adams Dr, Totowa, NJ 07512.

Publication date: September 1, 2004

More about this publication?
  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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