A high-throughput, simultaneous analysis of carotenoids, chlorophylls and tocopherol using sub two micron core shell technology columns

A high-throughput, robust and reliable method for simultaneous analysis of five carotenoids, four chlorophylls and one tocopherol was developed for rapid screening large sample populations to facilitate molecular biology and plant breeding. Separation was achieved for 10 known analytes and four unknown carotenoids in a significantly reduced run time of 10 min. Identity of the 10 analytes was confirmed by their UV-Vis absorption spectras. Quantification of tocopherol, carotenoids and chlorophylls was performed at 290 nm, 460 nm and 650 nm respectively. In this report, two sub two micron particle core-shell columns, Kinetex from Phenomenex (1.7 μm particle size, 12% carbon load) and Cortecs from Waters (1.6 μm particle size, 6.6% carbon load) were investigated and their separation efficiencies were evaluated. The peak resolutions were >1.5 for all analytes except for chlorophyll-a′ with Cortecs column. The ruggedness of this method was evaluated in two identical but separate instruments that produced CV < 2 in peak retentions for nine out of 10 analytes separated.