Genetics Discipline, Centre of Genomic Medicine Timișoara, "Victor Babeș" University of Medicine and Pharmacy, No 2, Eftimie Murgu Square, Timișoara 300041, Romania.
"Louis Țurcanu" Clinical Emergency Hospital for Children, No 2, Iosif Nemoianu St., Timișoara 300011, Romania.
Molecules. 2019 Jan 20;24(2):360. doi: 10.3390/molecules24020360.
Few high-performance liquid chromatography⁻tandem mass spectrometry (LC-MS/MS) methods have been developed for the full quantitation of fatty acids from human plasma without derivatization. Therefore, we propose a method that requires fewer sample preparation steps, which can be used for the quantitation of several polyunsaturated fatty acids in human plasma. The method offers rapid, accurate, sensitive, and simultaneous quantification of omega 3 (α-linolenic, eicosapentaenoic, and docosahexaenoic acids) and omega 6 fatty acids (arachidonic and linoleic acids) using high-performance LC-MS/MS. The selected fatty acids were analysed in lipid extracts from both free and total forms. Chromatographic separation was achieved using a reversed phase C18 column with isocratic flow using ammonium acetate for improving negative electrospray ionization (ESI) response. Mass detection was performed in multiple reaction monitoring (MRM) mode, and deuterated internal standards were used for each target compound. The limits of quantification were situated in the low nanomolar range, excepting linoleic acid, for which the limit was in the high nanomolar range. The method was validated according to the U.S. Department of Health and Human Services guidelines, and offers a fast, sensitive, and reliable quantification of selected omega 3 and 6 fatty acids in human plasma.
目前，尚未开发出无需衍生化即可对人血浆脂肪酸进行全定量分析的高效液相色谱-串联质谱（LC-MS/MS）方法。因此，我们提出了一种需要更少样品制备步骤的方法，可用于定量分析人血浆中的几种多不饱和脂肪酸。该方法采用高效 LC-MS/MS 对ω3（α-亚麻酸、二十碳五烯酸和二十二碳六烯酸）和 ω6 脂肪酸（花生四烯酸和亚油酸）进行快速、准确、灵敏和同时定量。所选择的脂肪酸以游离和总形式从脂质提取物中进行分析。采用反相 C18 柱，以乙酸铵作为流动相进行等度洗脱，以提高负离子电喷雾电离（ESI）响应，从而实现色谱分离。采用多重反应监测（MRM）模式进行质谱检测，并为每个目标化合物使用氘代内标。除亚油酸外，定量限均处于低纳摩尔范围，而亚油酸的定量限处于高纳摩尔范围。该方法根据美国卫生与公众服务部的指南进行了验证，可快速、灵敏、可靠地定量分析人血浆中的选定 ω3 和 ω6 脂肪酸。
