高效液相色谱法检测南极磷虾油中总虾青素含量  

Determination of Total Astaxanthin in Antarctic Krill Oil by High Performance Liquid Chromatography

作　　者：王松[1] 郝鹏飞[2] 赵鑫鹏[1] 苗钧魁[1] 刘小芳[1] 赵宪勇[1] 冷凯良[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所,山东青岛266071 [2]山东出入境检验检疫局,山东青岛266002

出　　处：《分析测试学报》2016年第4期482-486,共5页Journal of Instrumental Analysis

基　　金：青岛市博士后创新项目(ZQ51201415030);农业部项目"南极海洋生物资源开发利用"(1230135)

摘　　要：建立了一种检测南极磷虾油中总虾青素含量的高效液相色谱方法。样品采用Na OH-甲醇溶液皂化后加入Na HSO4终止反应,反应液经正己烷液液萃取净化,采用C18液相色谱柱分离,二极管阵列检测器在476 nm波长进行测定,虾青素在5 min内得到较好分析。考察了皂化时间对虾青素酯皂化效率的影响,研究结果显示,皂化30 min为最佳反应时间。虾青素含量在0.2-20.0 mg/L范围内与其峰面积呈良好的线性关系,相关系数为0.999 9,检出限为0.005 mg/kg,定量下限为0.2 mg/kg。日内精密度（RSD）≤3.4%,日间RSD≤4.3%,南极磷虾油样品的加标回收率为90.2%-95.2%。该方法的线性范围宽、准确性好、精密度高,样品前处理方法简便,适用于南极磷虾油中总虾青素含量的分析检测。A C18reversed-phase high performance liquid chromatographic（RP-HPLC） method was developed for the determination of total astaxanthin saponified from Antarctic krill oil.The sample was saponified with sodium hydroxide-methanol solution（4 mol / L）,the reaction was ended with sodium hydrogen sulfate,and the reaction liquid was purified by n-hexane liquid-liquid extraction.The effect of saponification time on saponification efficiency of astaxanthin in Antarctic krill oil was investigated.The results showed that the best saponification time was 30 min.Chromatographic separation was performed with a C18 liquid chromatographic column（250 mm × 4.6 mm,5 μm） at room temperature with an eluent of 100% methanol at a flow rate of 1.0 m L / min.The eluate was monitored with a diode array detector at 476 nm.The separation and identification of astaxanthin was completed within 5 min.The limits of detection（LOD） and quantitation（LOQ） for astaxanthin in Antarctic krill oil were 0.005 mg / kg and 0.2 mg / kg,respectively.A good linear relationship between chromatographic peak area and concentration in the range of 0.2-20.0 mg / L was obtained with a correlation coefficient of 0.999 9.The average recoveries were between 90.2% and 95.2%,and the inter-and intra-RSDs of the method were not more than 3.4% and 4.3%,respectively.The method was rapid,simple,precise and accurate,and could be applied in the inspection of astaxanthin in Antarctic krill oil.

关 键 词：虾青素 南极磷虾油 虾青素酯 皂化 高效液相色谱法 

分 类 号：O657.72[理学—分析化学] O629.4[理学—化学]