柱前衍生结合高效液相色谱-荧光检测法测定人体血浆中脂肪酸
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摘要
利用1-[1-(哌嗪)乙酮]-2-[(4-二甲氨基)苯基]-菲[9,10-d]咪唑(DPPIPE)作为柱前荧光标记试剂,建立了一种高灵敏度、高选择性的脂肪酸定量分析方法,并考察了方法的分析性能。通过对8种混合脂肪酸的优化衍生。采用Hypersil BDS C-8色谱柱(150×4.6 mm i.d.,5μm),以乙腈为流动相梯度洗脱,荧光检测激发和发射波长分别为260 nm和430 nm,对衍生物进行了基线分离以及定性定量分析。结果显示:线性回归系数多数大于0.9991,检测限0.290～0.593 pmol。在大气压化学电离源的正离子模式下,实现了人体血浆样品中脂肪酸组分的分离分析和鉴定。本方法操作简单、检测快捷、灵敏,对人体血浆样品的测定结果具有借鉴意义。
A simple and sensitive method for the determination of free fatty acids have been developed by high performance liquid chromatography(HPLC) with fluorescence detection using 1-[1-(piperazine) ethyl ketone]-2-[(4-dimethylamine) phenyl]-fi [9,10-d] imidazole(DPPIPE) as pre-column labeling reagent.On a reversed phased Hypersil BDS C-8 column,8 fatty acid derivatives were separated and analyzed quantitatively.Fluorescent detection excitation and emission wavelengths were set at λ_(ex)=260 nm and λ_(em)=430 nm,respectively.Correlation coefficients for fatty acid derivatives were more than 0.9991,and detection limits(at signal-to-noise of 3∶1) were in the range of 0.290-0.593 pmol.Free fatty acids from human plasma samples were identified by the on line MS equipped with an atmospheric pressure chemical ionization source.When applying to the determination of free fatty acids from real samples,this developed method exhibited excellent sensitivity and reproducibility.
A simple and sensitive method for the determination of free fatty acids have been developed by high performance liquid chromatography(HPLC) with fluorescence detection using 1-[1-(piperazine) ethyl ketone]-2-[(4-dimethylamine) phenyl]-fi [9,10-d] imidazole(DPPIPE) as pre-column labeling reagent.On a reversed phased Hypersil BDS C-8 column,8 fatty acid derivatives were separated and analyzed quantitatively.Fluorescent detection excitation and emission wavelengths were set at λ_(ex)=260 nm and λ_(em)=430 nm,respectively.Correlation coefficients for fatty acid derivatives were more than 0.9991,and detection limits(at signal-to-noise of 3∶1) were in the range of 0.290-0.593 pmol.Free fatty acids from human plasma samples were identified by the on line MS equipped with an atmospheric pressure chemical ionization source.When applying to the determination of free fatty acids from real samples,this developed method exhibited excellent sensitivity and reproducibility.
引文
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