高效液相色谱/质谱联用直接测定鱼油中EPA/DHA含量

Determination of eicosapentaenoic acid and docosahexaenoic acid infish oil by high performance liquid chromatography/mass spectrometry

建立快速、简单、准确测定鱼油样品中EPA和DHA两种ω-3PUFA含量的高效液相色谱/电喷雾质谱联用(HPLC-ESI/MS)分析方法。鱼油经2mol/LNaOH乙醇溶液皂化、3mol/LHCL酸化后,使用Waters液相色谱仪、SymmetryC8柱(2.1×150mm),以甲醇—水为流动相、十七酸为内标,利用质谱定性定量测定EPA/DHA含量。质谱在电喷雾负离子模式下,对m/z301、m/z327和m/z269进行选择离子监测。该方法下EPA、DHA分别在5.55～55.50μg/mL和0.90～9.00μg/mL范围内峰面积和浓度呈良好的线性关系,相关系数REPA=0.9995、RDHA=0.9993;EPA、DHA的回收率分别为98.63%～99.23%、97.12%～99.17%,EPA、DHA的相对标准偏差分别为1.57%、1.20%(n=5)。此方法流动相简单,分析时间短且无需衍生处理,不受色谱分离度的限制并可实现对样品的重复分析,能准确快速测定鱼油中EPA和DHA含量。

HPLC-ESI/MS was established to determine the content of EPA and DHA in fish oil.The fish oil was saponified with 2 mol/L NaOH-C2H5OH,acidified with 3 mol/LHCL until slightly acidic,and injected directly into the system of HPLC.The analysis was carried out on SymmetryC8(2.1×150 mm)column.C17:0 was selected as internal standard for quantification.EPA and DHA was identified by electrospray ionization mass spectrometry(ESI-MS),and electrospray was operated in negative ion mode to generate protonated ions.The io...