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United States Department of Agriculture

Agricultural Research Service

Research Project: Dietary Modulation of Immune Function and Oxidative Stress

Location: Immunity and Disease Prevention Research Unit

Title: Rapid isocratic HPLC method and sample extraction procedures for measuring carotenoid, retinoid, and tocopherol concentrations in human blood and breast milk for intervention studies

Authors
item Turner, Tami -
item Burri, Betty

Submitted to: Chromatographia
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: January 25, 2012
Publication Date: February 10, 2012
Repository URL: http://www.springerlink.com/content/b256868h7624m316/fulltext.pdf
Citation: Turner, T., Burri, B.J. 2012. Rapid isocratic HPLC method and sample extraction procedures for measuring carotenoid, retinoid, and tocopherol concentrations in human blood and breast milk for intervention studies. Chromatographia. 75:241-252. DOI 10.1007/s10337-012-2193-9.

Interpretive Summary: Vitamin A deficiency is an important cause of death in the developing world. Programs to alleviate vitamin A deficiency by feeding people provitamin A-carotenoid rich fruits and vegetables are being tested now in several countries. They need a simple, fast method to measure vitamin A and provitamin A carotenoids in blood and breast milk. We developed a new HPLC method for these measurements. It is a reversed-phase method using 7:2:1 acetonitrile: dichloromethane: methanol at 0.5ml/min. Analysis is complete in approximately eight minutes. This rapid method uses little solvent and provides excellent results in the analysis of these nutrients. These methods provide efficient analysis of carotenoids and retinoids for large quantities of blood and breast milk.

Technical Abstract: Vitamin A deficiency is an important cause of death in the developing world. A variety of dietary interventions using provitamin A-carotenoid rich fruits and vegetables are being tested for their potential to alleviate vitamin A deficiency in several countries. Thus, there is a need for a simple, fast isocratic reverse-phase HPLC method to analyze vitamin A and provitamin A carotenoids in biological samples. An Agilent 1100 system equipped with a Waters Spherisorb ODS2 column (3x125mm 3µm) and similar guard column (ThermoScientific ODS2 3x20mm 3µm) was used. Mobile phase of 7:2:1 acetonitrile: dichloromethane: methanol was run at 0.5ml/min. Run times were complete in approximately eight minutes. This rapid method uses little solvent and provides excellent results in the analysis of these fat-soluble nutrients. Repeatability was 1.0 - 5.1% (RSD), and limits of detection 1.3 - 15 ng/ml. Several extraction methods for carotenoids and retinol in plasma and human breast milk were also performed for efficiency comparisons. These methods provide efficient analysis of carotenoids and retinoids for large quantities of blood and breast milk.

Last Modified: 12/21/2014
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