Metabolomic Approach for Carotenoids Analysis of Chrysanthemum fower by Liquid Chromatograph Tandem Mass Spectrometry

Significance of the Topic

Carotenoids are key pigments determining flower color and market value in ornamental crops such as Chrysanthemum morifolium. A comprehensive metabolomic analysis of these compounds enables rapid screening of breeding lines, supports quality control, and advances our understanding of biosynthetic pathways that underlie color variation.

Objectives and Study Overview

This study aimed to establish a metabolomic platform for simultaneous profiling of major and minor carotenoids in chrysanthemum flowers. By comparing wild‐type and radiation‐induced mutant lines, researchers sought to identify alterations in carotenoid composition and link them to changes in flower color, thereby facilitating efficient breeding strategies.

Methodology and Instrumentation

The analytical workflow combined a straightforward hexane extraction with high‐resolution chromatographic separation and targeted mass spectrometry detection.

• Sample Preparation and Extraction: 10 mg of freeze‐dried flower powder was sonicated in n‐hexane, centrifuged, and the supernatant used for analysis.
• Chromatographic Separation: Reversed‐phase UHPLC on a SunShell C18 column (2.1 × 50 mm, 2.6 µm) with a gradient from 70% to 100% organic (acetonitrile/2-propanol).
• Mass Spectrometric Detection: Shimadzu LCMS-8050 triple quadrupole with APCI source operated in SIM and MRM modes, optimized for 17 carotenoid standards.

Used Instrumentation

• Shimadzu Nexera UHPLC system with SunShell C18 column (2.1 × 50 mm, 2.6 µm).
• Shimadzu LCMS-8050 triple quadrupole mass spectrometer with APCI (±), spray voltages +4.5 kV/−3.5 kV, nebulizing gas 3 L/min, drying gas 5 L/min, interface 350 °C, DL 200 °C, heater block 300 °C.

Main Results and Discussion

Chromatograms revealed clear differences between wild‐type and mutant flowers. In yellow mutants, levels of zeinoxanthin were markedly reduced, while a lutein‐related epoxide ((3S,5S,6R,3′R,6′R)‐5,6‐dihydro‐5,6‐dihoxylutein) accumulated. Metabolic pathway mapping indicated that radiation‐induced mutations affect hydroxylation and epoxidation steps from zeinoxanthin to lutein epoxide. Cluster heatmap analysis across multiple breeding lines highlighted groups of mutants with distinct carotenoid signatures, demonstrating the platform’s potential for high‐throughput screening.

Benefits and Practical Applications

This targeted LC‐MS/MS approach allows rapid, sensitive, and reproducible quantification of diverse carotenoids in flower tissues. It supports breeding programs by linking specific metabolite profiles to desired color traits, enables quality assurance in nurseries, and can be adapted to other ornamental and crop species for trait improvement.

Future Trends and Potential Uses

Advancements may include integration with automated sample preparation and high‐throughput phenotyping, expansion to untargeted metabolomics for discovery of novel pigments, and application of machine learning for pattern recognition in large breeding populations. Extending this platform to multi‐omics studies will deepen insights into regulatory networks governing pigment biosynthesis.

Conclusion

A robust, targeted LC-MS/MS metabolomic workflow was developed for simultaneous profiling of over a dozen carotenoids in chrysanthemum flowers. The method discriminates wild-type and mutant lines, uncovers pathway perturbations caused by radiation breeding, and provides a practical tool for accelerating color trait selection in ornamental horticulture.

Reference

1. Omiya A. Diversity of Carotenoid Composition in Flower Petals. JARQ. 2011;45(2):163–171.