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目的建立测定母乳中四种主要多不饱和脂肪酸(PUFA)的高效液相色谱法(HPLC)。方法用(甲醇:正己烷:1mol/L磷酸=15:25:3)提取,ω-溴代苯乙酮衍生化,流动相为(乙腈:水=81:19,v/v),流速1.0 ml/min,柱温35℃,分析柱依利特Hypersil BDS C18(4.6mm×150mm,5μm),紫外检测波长为252 nm。结果α-亚麻酸(18:3n-3,α-LNA)、二十二碳六烯酸(22:6n-3,DHA)、花生四烯酸(20:4n-6,AA)和亚油酸(18:2n-6,LA)可以完全分离,峰形良好。平均回收率均在(98±5)%以内,提取回收率均≥75%。各PUFA质控母乳的日内RSD≤4.6%,日间RSD≤5.9%。结论本方法稳定、可靠,适用于母乳中PUFA浓度测定。
Objective To establish a HPLC method for the determination of four major polyunsaturated fatty acids (PUFA) in breast milk. The method was extracted with (methanol: n-hexane: 1mol / L phosphoric acid = 15: 25: 3) and derivatized with ω-bromoacetophenone. The mobile phase was acetonitrile: water = 81: 19, v / v. ml / min and the column temperature was 35 ℃. The analytical column was Hypersil BDS C18 (4.6mm × 150mm, 5μm). The UV detection wavelength was 252 nm. Results The results showed that α-linolenic acid (18: 3n-3, α-LNA), docosahexaenoic acid (22: 6n-3, DHA), arachidonic acid Acid (18: 2n-6, LA) can be completely separated, the peak shape is good. The average recovery was within (98 ± 5)%, the extraction recovery was ≥75%. Each PUFA quality control breast milk RSD ≤ 4.6%, daytime RSD ≤ 5.9%. Conclusion The method is stable and reliable and suitable for the determination of PUFA concentration in breast milk.