Abstract
Separation of analytes is an important factor affecting the effectiveness of carotenoids analysis from a complex extraction. In this study, a high performance liquid chromatography procedure with improved separation was developed to determine carotenoids extracted from P. rhodozyma. Mixture design experiment was employed to optimize the organic solvent constitution, which showed that the optimal organic phase was composed (v/v) of 61% of methanol, 32% of tetrahydrofuran, and 7% of acetonitrile. Furthermore, the gradient elution procedure optimization demonstrated that the gradient elution started at the optimal organic phase concentration of 60% lead to the best separation effect. Meanwhile, 18 compounds in total 25 detected peaks were separated. Based on their absorption spectra, astaxanthin, s-carotene, zeaxanthin, β-cryptoxanthin and lycopene were identified by matching their retention times with standards. Interestingly, our improved HPLC method was effective to classify P. rhodozyma strains and monitor the degradation of astaxanthin in accordance with their carotenoids composition.
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