An improved method for fast and selective separation of carotenoids by LC–MS

Daniel Abate-Pella, Dana M. Freund, Janet P. Slovin, Adrian D. Hegeman, Jerry D. Cohen

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Carotenoids are a large class of compounds that are biosynthesized by condensation of isoprene units in plants, fungi, bacteria, and some animals. They are characteristically highly conjugated through double bonds, which lead to many isomers as well susceptibility to oxidation and other chemical modifications. Carotenoids are important because of their potent antioxidant activity and are the pigments responsible for color in a wide variety of foods. Human consumption is correlated to many health benefits including prevention of cancer, cardiovascular disease, and age-related disease. Extreme hydrophobicity, poor stability, and low concentration in biological samples make these compounds difficult to analyze and difficult to develop analytical methods for aimed towards identification and quantification. Examples in the literature frequently report the use of exotic stationary phases, solvents, and additives, such as ethyl acetate, dichloromethane, and methyl tert-butyl ether that are incompatible with liquid chromatography mass spectrometry (LC–MS). In order to address these issues, we implemented the use of LC–MS friendly conditions using a low-hydrophobicity cyano-propyl column (Agilent Zorbax SB-CN). We successfully differentiated between isomeric carotenoids by optimizing two gradient methods and using a mixture of 11 standards and LC–MS in positive ionization mode. Three complex biological samples from strawberry leaf, chicken feed supplement, and the photosynthetic bacterium Chloroflexus aurantiacus were analyzed and several carotenoids were resolved in these diverse backgrounds. Our results show this methodology is a significant improvement over other alternatives for analyzing carotenoids because of its ease of use, rapid analysis time, high selectivity, and, most importantly, its compatibility with typical LC–MS conditions.

Original languageEnglish (US)
Pages (from-to)34-37
Number of pages4
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume1067
DOIs
StatePublished - Nov 1 2017

Bibliographical note

Funding Information:
This work was supported by the National Science Foundation [grant numbers IOS-1238812 , IOS-1400818 ]. We thank Joseph Kuo-Hsiang Tang for providing the C. aurantiacus samples and Frederick Khachik for carotenoid standards.

Publisher Copyright:
© 2017 Elsevier B.V.

Keywords

  • Carotenoids
  • Cyano-propyl column
  • LC–MS

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