Skip to main content
ARS Home » Northeast Area » Beltsville, Maryland (BHNRC) » Beltsville Human Nutrition Research Center » Methods and Application of Food Composition Laboratory » Research » Publications at this Location » Publication #427872
Research Project: Farm to Table Factors: Impact of Production, Processing, and Preparation on Food Composition

Location: Methods and Application of Food Composition Laboratory

Title: Quantification of carotenoids in foods using relative response factor by HPLC with photodiode array detector

Author
item LEE, ETHAN - University Of Maryland
item YU, LIANGLI - University Of Maryland
item Wu, Xianli

Submitted to: ACS Food Science and Technology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/9/2025
Publication Date: 10/20/2025
Citation: Lee, E.Y., Yu, L., Wu, X. 2025. Quantification of carotenoids in foods using relative response factor by HPLC with photodiode array detector. ACS Food Science and Technology. 5(11):4301-4309. https://doi.org/10.1021/acsfoodscitech.5c00741.
DOI: https://doi.org/10.1021/acsfoodscitech.5c00741

Interpretive Summary: Carotenoids are naturally occurring plant pigments and play important roles in human health. Quantification of carotenoids in plant-based foods is normally performed using HPLC or HPLC-MS with individual standards. However, low stability and availability of carotenoid standards remain a key challenge to the accurate quantification of carotenoids. In this study a quantification method using relative response factors (RRF) in HPLC-PDA was developed and validated for quantitation of carotenoids. Authentic standards of 10 carotenoids commonly found in plant-based foods were obtained and their purities were determined. RRF of each individual standard was generated. Method validation was conducted. In addition, quantifications of carotenoids in selected food samples by HPLC using RRF and by using individual standards were conducted and compared. Method validation results suggested that this method is reliable and reproducible. Quantifications of carotenoids using RRFs yielded comparable and consistent results when compared to those using individual standards. In conclusion, this method provides an accurate, consistent, and cost-effective approach in quantifying major carotenoids in plant foods.

Technical Abstract: Introduction: Carotenoids are naturally occurring plant pigments and play important roles in human health. Quantification of dietary carotenoids is normally performed using HPLC or HPLC-MS with individual standards. However, low stability and availability of carotenoid standards remain a key challenge to the accurate quantification of carotenoids. Objective: A quantification method using relative response factors (RRF) in HPLC-PDA was developed and validated for quantitation of carotenoids in plant foods. Materials and Methods: Authentic standards of 10 carotenoids commonly found in plant-based foods were obtained commercially. RRF of each individual standard was generated against ß-carotene. Method validation was conducted to assess specificity, limit of detection, limit of quantification, the linearity and working range, precision and accuracy. In addition, quantifications of carotenoids in selected food samples by HPLC using RRF and by using individual standards were conducted and compared. Results: Method validation results suggested the method is reliable and reproducible. Quantifications of carotenoids in foods using RRFs yielded comparable and consistent results when compared to those using individual standards. Conclusion: The quantification method using RRF provides an accurate, consistent, and cost-effective approach in quantifying major carotenoids in plant foods.