柱前衍生-高效液相色谱荧光检测法测定高寒沼泽牧草及藻体中脂肪酸含量

Analysis of fatty acids in herbage and alga of alpine marshes by pre-column derivation coupled-high performance liquid chromatography with fluorescence detection

为了研究牧草及藻体内脂肪酸组成的差异,本研究以N-乙基-N,-[(3-二甲氨基)丙基]碳二亚胺盐(EDC)的乙腈溶液为催化剂,采用新型荧光试剂10-甲基吖啶酮-2-磺酰哌嗪(MASP)作为柱前衍生试剂,对19种混合脂肪酸进行优化衍生,以λ_(ex)=315 nm和λ_(em)=430 nm分别为荧光检测的激发和发射波长,采用梯度洗脱,实现了对19种混合脂肪酸的完全基线分离及定量分析.结果显示,多数脂肪酸的线性回归系数大于0.99,且检出限在17.6~42.7 ng·mL^(-1)(S/N=3∶1).上述方法具有良好的重现性,能够满足高寒沼泽牧草及藻体样品中脂肪酸测定的需要.

To study the differences in fatty acid composition between herbage and algae,19 kinds of mixed fatty acids(FA)were optimized and derived by using new fluorescent reagent 10-methyl acidone-2-sulfonyl piperazine(MASP)as pre-column derivative reagent.In the study,the N-ethyl-N,-((3-dimethylamino)propyl)acetonitrile solution of carbon diimide salt(EDC)was used as acatalyst.After sealing the derivative reaction 60 min at 60℃,a stable fluorescence product was obtained by using the hypersil BDS C18(4.6 mm×100 mm,5μm)chromatographic column,gradient elution and excitation wavelengthsλ_(ex)=315 nm and emission wavelengthsλ_(em)=430 nm.Baseline separation and quantitative analysis of 19 kinds of fatty acids(FA)were achieved.The experimental results showed that the linear regression coefficient of most fatty acids is greater than 0.99 and the limit of detection is 17.6 to 42.7 ng·mL^(-1)(S/N=3∶1).The above experimental method has good reproducibility and can satisfy the needs for the determination of fatty acid status of herbage and algae samples in alpine swamps.