GPC-HPLC法测定南极磷虾油中虾青素及校正因子计算  

Determination of astaxanthin in antarctic krill oil by GPC-high performance liquid chromatography and calculation of correction factor

作　　者：孙伟红[1] 冷凯良[1] 邢丽红[1] 朱敏[2] 翟毓秀[1] 苗均魁[1] 朱兰兰[1] 

机构地区：[1]中国水产科学研究院黄海水产研究所,青岛266071 [2]中国海洋大学食品科学与工程学院,青岛266100

出　　处：《分析试验室》2013年第2期26-30,共5页Chinese Journal of Analysis Laboratory

基　　金：国家高技术研究发展计划项目(2011AA090801)资助

摘　　要：以保健食品南极磷虾油为研究对象,建立了测定南极磷虾油中虾青素含量的全自动凝胶渗透色谱(GPC)净化-高效液相色谱分析方法。样品采用GPC净化,NaOH甲醇溶液皂化后,经C30液相色谱柱分离后测定。考察了皂化溶剂、加碱量、皂化时间等对虾青素皂化效率的影响以及pH对溶液稳定性的影响,并对虾青素异构体的校正因子进行了计算,确定了虾青素的定量方式。南极磷虾油中虾青素的定量限为0.5 mg/kg;在0.1～5 mg/L时峰强度与质量浓度的线性关系良好(r>0.999);加标回收率为96%～98.5%;相对标准偏差为3.0%～5.6%。方法适用于南极磷虾油中虾青素的实际检测。Taking health products, antarctic krill oil, as the object in the paper, an analytical method based on gel permeation chromatography-high performance liquid chromatography （GPC-HPLC） has been developed for the determination of astaxanthin in antarctic krill oil. The sample was purified by gel permeation chromatography, saponified by sodium hydroxide-methanol solution, and separated by a C30 liquid chromatographic column. In this paper, we established the purification conditions of gel permeation chromatography, the collection time of astaxanthin and astaxanthin easter was 7.48 - 12.60 min. The conditions of saponification were researched by exploring the solvents, volume of sodium hydroxide-methanol solution and saponification time on saponification efficiency of astaxanthin in antarctic krill oil. The results show that methylene chloride： methanol as saponification solvent had the best effect than other solvents, the most suitable volume for 0.2 mol/L sodium hydroxide-methanol solution was 1 mL, and the best saponification time was 12 h by comparing among 1 - 18h at 4 ~C. We also investigated the effect of pH on stability of astaxanthin in antarctic krill oil, the content of astaxanthin had less influence on neutral environment among 0 -12 h, but the content in serious decline and isomerization on alkaline environment. The contents of isomers of astaxanthin in antarctic krill oil were calculated by calibration factor, the calibration factor of 13-cis-astaxanthin to trans-astaxanthin was 1.3, and 9-cis-astaxanthin to trans-astaxanthin was 1.1, so we got the quantitative method of astaxanthin in krill oil. The limits of quantification for astaxanthin in antarctic krill oil were 0.5 mg/kg. There were good linear relationship between the chromatographic peak area and the concentration in the range of 0. 1 mg/L -5 mg/L with correlation coefficient over 0. 999. The average recoveries were between 96.0% and 98.5%, and the precision of the method were 3.0% -5.6%. The method is very simple, convenient, accurate and

关 键 词：南极磷虾油 虾青素 凝胶色谱净化 液相色谱法 校正因子. 

分 类 号：O657.7[理学—分析化学]