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cis/trans Carotenoid Extraction, Purification, Detection, Quantification, and Profiling in Plant Tissues
Yagiz Alagoz1, Namraj Dhami1, Chris Mitchell1
1Hawkesbury Institute for the Environment, Western Sydney University, Locked Bag 1797, Penrith, 2751, NSW, Australia.
Methods in Molecular Biology (Clifton, N.J.)
|November 21, 2019
Summary
Optimized high-performance liquid chromatography (HPLC) methods simplify carotenoid analysis in plants. These methods improve the identification and quantification of various carotenoid isomers, enhancing research accuracy.
Area of Science:
- Plant Biochemistry
- Analytical Chemistry
- Chromatography
Background:
- Reverse phase high-performance liquid chromatography (HPLC) is crucial for analyzing carotenoids in biological, health, and food research.
- Carotenoid isomer identification and quantification are sensitive to sample preparation, extraction, HPLC column chemistry, and solvent gradients.
- Existing methods exhibit significant variability in ease of use, efficiency, and accuracy.
Purpose of the Study:
- To present a simplified carotenoid extraction protocol.
- To describe two optimized HPLC methods using C18 and C30 reverse-phase columns for carotenoid analysis.
- To provide guidance on profiling and quantifying carotenoids in diverse plant tissues.
Main Methods:
- Development of a simple carotenoid extraction technique.
- Optimization of two distinct HPLC methods employing C18 and C30 reverse-phase columns.
- Characterization of spectral properties and retention times for individual carotenoid species.
Main Results:
- Successful application of optimized HPLC methods for detecting xanthophylls and cis-carotenes.
- Demonstrated utility in both wild-type photosynthetic leaves and mutant dark-grown etiolated seedlings.
- Provided resources for comprehensive carotenoid profiling and quantification.
Conclusions:
- The described methods offer improved accuracy and efficiency for carotenoid analysis.
- These optimized HPLC protocols are valuable tools for plant research, enabling detailed profiling and quantification.
- The study addresses the heterogeneity in existing methods by providing standardized, effective approaches.
