三角褐指藻中岩藻黄素的分离纯化工艺研究

Isolation and purification of fucoxanthin from Phaeodactylum tricornutum

目的 探索三角褐指藻(Phaeodactylum tricornutum)冻干藻粉中岩藻黄素的分离及纯化方法,为深入研究三角褐指藻中岩藻黄素的工业化制备提供技术方法。方法 培养三角褐指藻并获得冻干藻粉,70%乙醇进行提取,ODS开放柱层析分离,乙醇沉淀法进行纯化得到岩藻黄素。用电喷雾质谱法(ESI-MS)和核磁共振波谱法(1H NMR)对纯化得到的岩藻黄素进行结构表征。结果 ODS柱层析分离过程中,绝大多数岩藻黄素在80%乙醇条件下成功洗脱,回收率为(88.02±2.30)%。乙醇沉淀的最佳工艺为醇沉含醇量50%、4℃静置24 h。在此乙醇析出条件下,岩藻黄素的回收率为(86.84±0.99)%,纯度为(95.79±1.06)%。分离纯化得到的岩藻黄素经质谱和核磁共振波谱鉴定,其结构为全反式岩藻黄素。结论 该工艺为全流程绿色高效分离纯化技术,工艺简单,岩藻黄素收率高,为进一步研究以三角褐指藻为原料工业化生产高纯度岩藻黄素提供了技术方法,为岩藻黄素产品在食品和医药领域的开发提供了理论依据。

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%(v/v)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(v/v)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 24h.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.