Separation and Identification of Lutein, β-Cryptoxanthin and Their Isomers
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Abstract:
A method to simultaneously detect all-trans-Lutein, all-trans-β-cryptoxanthin and their stereoisomers was developed using C30-column, high-performance liquid chromatography, diode array (C30-HPLC-DAD) technique. The structures of isolated compounds were elucidated by atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) and ultraviolet/visible (UV/Vis) spectroscopy. The optimal chromatographic condition was as follows: mobile phase A: 1.5% ammonium acetate:water:methyl tert-butyl ether (MTBE)-MeOH (5:25:70, V/V/V), mobile phase B: 1.5% ammonium acetate:water:MTBE-MeOH (5:85:10, V/V/V); flow rate: 0.6 mL/min; elution time: 24 min; injection volume: 20 μL; column temperature: 25 ℃. Using this novel method, 15-cis-lutein, 13/13′-cis-lutein, 9-cis-lutein, 9′-cis-lutein, 15-cis-β-cryptoxanthin, 13- and 13′-cis-β-cryptoxanthin, 9-cis-β-cryptoxanthin and 9′-cis-β-cryptoxanthin were successfully separated from the mixture. The peak areas of all-trans lutein and all-trans β-cryptoxanthin showed a good linear relationship with their injection volumes in the range of 2 to 150 ng and 5 to 250 ng, respectively. The proposed method accurately and rapidly separated all-trans lutein, all-trans β-cryptoxanthin, and their cis-isomers simultaneously. This method has potential applications in qualitative and quantitative analysis of all-trans lutein, all-trans β-cryptoxanthin, and their stereoisomers in fruits and vegetables.
