高效液相法测定海洋硅藻中5种叶黄素类化合物

Determination of Five Xanthophylls in Marine Diatoms by High Performance Liquid Chromatography

建立了运用高效液相法同时测定海洋硅藻中5种叶黄素类化合物的方法。采用Agilent Eclipse XDB-C18色谱柱为固定相,以乙腈和水为流动相,对新黄素、岩藻黄素、硅甲藻黄素、硅藻黄素、玉米黄素的含量进行了测定。结果表明,新黄素、岩藻黄素、硅甲藻黄素在0.2~20 mg/L,硅藻黄素在2.0~20 mg/L,玉米黄素在5.0~20 mg/L范围内的线性关系良好,相关系数(r2)均高于0.999,回收率在88.40%~115.00%,保留时间的日内和日间精密度的相对标准偏差(RSD)分别为0.13%~0.68%和0.13%~1.15%;峰面积的日内和日间精密度的相对标准偏差(RSD)分别为0.33%~3.56%和1.07%~4.97%。方法的检出限(LOD)在0.06~1.00 mg/L,定量限(LOQ)在0.20~3.00 mg/L。利用该方法对8种硅藻藻株中5种叶黄素类化合物的含量进行分析,比较了三角褐指藻、角毛藻、中肋骨条藻、舟形藻、双眉藻和梅尼小环藻之间的5种叶黄素类化合物的分布情况。该方法具有操作简单、灵敏度高、重复性好、回收率高等优点,适合硅藻中多种叶黄素类化合物的同时测定,为海洋硅藻的藻种优选、培养条件优化、制备技术开发提供了分析手段。

A quantitative method based on high performance liquid chromatography(HPLC)has been established for five xanthophylls in marine diatom.The HPLC method used a reversed phase Agilent Eclipse XDB-C 18 column.Acetonitrile and water were used as mobile phases.Five xanthophylls,including neoxanthin,fucoxanthin,diadinoxanthin,diatoxanthin and zeaxanthin,were quantified in this study.The results indicated that the linearities of neoxanthin,fucoxanthin and diadinoxanthin were good in the ranges of 0.2~20 mg/L,while the linearities of diatoxanthin and zeaxanthin were good in the ranges of 2.0~20 mg/L and 5.0~20 mg/L,respectively.And their correlation coefficients(r 2)were higher than 0.999.The recoveries of the five xanthophylls were between 88.40%and 115.00%.The inter-day and intra-day precisions(RSD)of retention time were 0.13%~0.68%and 0.13%~1.15%,respectively.The inter-day and intra-day precisions(RSD)of peak area were 0.33%~3.56%and 1.07%~4.97%,respectively.The detection limits of the five xanthophylls for this method were between 0.06 and 1.00 mg/L and the quantification limits were in the range from 0.20 to 3.00 mg/L.According to the above method,eight diatom strain,include Phaeodactylum tricornutum Bahl.(NMBguh001),Chaetoceros sp.(NMBguh003-3),Chaetoceros sp.(NMBguh003-11),Chaetoceros sp.(NMBguh003-17),Skeletonema costatum(NMBguh004-2),Navicula sp.(NMBguh006-6),Amphora sp.(NMBguh007),Cyclotella meneghiniana(NMBguh026-8),have been investigated for comparing the xanthophylls distributions.The method has the advantages of simple,sensitive,accurate,reproducible and recovery rate.It is suitable for the simultaneous determination of various xanthophylls in diatoms,and provides an analytical means for the screen of algal species,the optimization of culture conditions and preparation technology of marine diatoms.