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Peer-reviewedCC BY 4.0Molecules

A Five-Minute UHPLC–MS/MS Method Facilitates Anthocyanin Biofortification in Maize (Zea mays L.)

  • Franz J. O’Hanna1
  • Shaji K. Chacko1
  • Samuel D. Jesse1
  • Alexander I. Hernandez1
  • Wenwei Xu2
  • Timothy Paape2,3
  • Michael P. Dzakovich1
  1. 1USDA-ARS Children’s Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 1100 Bates Ave., Houston, TX 77030, USA
  2. 2Department of Soil and Crop Sciences, Texas A&M University, 370 Olsen Blvd., College Station, TX 77843, USA
  3. 3USDA-ARS Responsive Agriculture Food Systems Research Unit, 498 Olsen Blvd., College Station, TX 77843, USA
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Abstract

Anthocyanin-rich maize offers opportunities for nutritional biofortification and natural food colorants, but conventional chromatographic methods limit the throughput needed for large-scale breeding programs. We developed a 5-minute ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method for quantifying maize anthocyanins and compared its performance with a 25-minute method. Anthocyanins from five commercial products and one breeding line were analyzed using optimized chromatographic gradients and multiple-reaction monitoring. Anthocyanin concentration and composition varied substantially among varieties, with differing contributions from cyanidin-, peonidin-, and pelargonidin-derived compounds. The 5-minute method reduced analytical runtime fivefold while preserving the relative ranking of varieties by total anthocyanin concentration, although absolute concentrations differed slightly between methods for most varieties. Random-effects modeling indicated that the methodology used to phenotype maize represented less than 1% of the total variation, further reinforcing that our rapid method is suitable for profiling breeding populations. This rapid UHPLC–MS/MS method provides a high-throughput analytical platform for quantifying predominant maize anthocyanins, facilitating phenotyping and selection for anthocyanin biofortification in maize breeding programs.

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Cite

BibTeXRIS
BibTeX
@article{ohanna2026fiveminute,
  title = {{A Five-Minute UHPLC–MS/MS Method Facilitates Anthocyanin Biofortification in Maize (Zea mays L.)}},
  author = {Franz J. O’Hanna and Shaji K. Chacko and Samuel D. Jesse and Alexander I. Hernandez and Wenwei Xu and Timothy Paape and Michael P. Dzakovich},
  journal = {Molecules},
  year = {2026},
  month = oct,
  doi = {10.3390/molecules31213920},
  url = {https://www.mdpi.com/1420-3049/31/21/3920},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - A Five-Minute UHPLC–MS/MS Method Facilitates Anthocyanin Biofortification in Maize (Zea mays L.)
AU  - Franz J. O’Hanna
AU  - Shaji K. Chacko
AU  - Samuel D. Jesse
AU  - Alexander I. Hernandez
AU  - Wenwei Xu
AU  - Timothy Paape
AU  - Michael P. Dzakovich
PY  - 2026
DA  - 2026/10/30
JO  - Molecules
DO  - 10.3390/molecules31213920
UR  - https://www.mdpi.com/1420-3049/31/21/3920
AB  - Anthocyanin-rich maize offers opportunities for nutritional biofortification and natural food colorants, but conventional chromatographic methods limit the throughput needed for large-scale breeding programs. We developed a 5-minute ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) method for quantifying maize anthocyanins and compared its performance with a 25-minute method. Anthocyanins from five commercial products and one breeding line were analyzed using optimized chromatographic gradients and multiple-reaction monitoring. Anthocyanin concentration and composition varied substantially among varieties, with differing contributions from cyanidin-, peonidin-, and pelargonidin-derived compounds. The 5-minute method reduced analytical runtime fivefold while preserving the relative ranking of varieties by total anthocyanin concentration, although absolute concentrations differed slightly between methods for most varieties. Random-effects modeling indicated that the methodology used to phenotype maize represented less than 1% of the total variation, further reinforcing that our rapid method is suitable for profiling breeding populations. This rapid UHPLC–MS/MS method provides a high-throughput analytical platform for quantifying predominant maize anthocyanins, facilitating phenotyping and selection for anthocyanin biofortification in maize breeding programs.
SN  - 1420-3049
ER  -