Objective To explore the method for isolation and purification of fucoxanthin from freeze-dried biomass of Phaeodactylum tricornutum and to provide technical methods for further research on the production of fucoxanthin from P.tricornutum in industrial scale.Methods The diatom P.tricornutum were cultivated and freeze-dried.The freeze-dried P.tricornutum was extracted with 70％ethanol.Then ODS open column chromatography was used for fucoxanthin separation.Then fucoxanthin was purified by ethanol precipitation method.The structure of purified fucoxanthin was determined by electrospray ionization mass spectrometric(ESI-MS)and 1H NMR data.Results The majority of fucoxanthin was successfully eluted by 80％ethanol/water with a recovery of(88.02±2.30)％using ODS column chromatography.The optimum ethanol precipitation were as follows:ethanol content 50％,precipitation temperature 4 ℃ and precipitation duration 24 h.Under the ethanol precipitation conditions,the recovery rate of fucoxanthin was(86.84±0.99)％,and the purity was(95.79±1.06)％.The purified fucoxanthin was determined as all-trans-fucoxanthin by mass spectrometry and nuclear magnetic resonance spectroscopy.Conclusion This study provided a green and efficient isolation and purification technology for the whole process with the advantages of simple process and high yield of fucoxanthin.It provided a technological method for the further research of high-purity fucoxanthin production from P.tricornutum in industrial-scale,and offered a theoretical basis for the development of fucoxanthin products in food and medicine.
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