Bailey-Hall Eileen, Nelson Edward B, Ryan Alan S
Martek Biosciences Corporation, Columbia, MD 21045, USA.
Lipids. 2008 Feb;43(2):181-6. doi: 10.1007/s11745-007-3140-7. Epub 2007 Dec 15.
An assay involving a finger stick and filter paper blood spotting was developed to determine polyunsaturated fatty acid (PUFA) levels in blood. Capillary whole blood from a finger stick was blotted on antioxidant impregnated filter paper, air dried, saponified and methylated using sodium hydroxide and boron trifluoride in methanol. The method differed from those described previously because separation of plasma and red blood cells (RBCs) was not needed, thin-layer chromatography (TLC) was not required to separate phospholipids, initial extraction of lipids before transesterification was not necessary, and the fatty acid methyl ester (FAME) method was able to methylate steryl esters, free fatty acids, and sphingomyelins. Twenty-six subjects provided blood samples by finger stick and venipuncture. Levels of long-chain polyunsaturated fatty acids (LC-PUFA) from capillary whole blood were correlated with those from RBCs and PLs in venous blood (P < 0.001, R(2) ranged from 0.64 to 0.86). Although highly significant (P < 0.002), the R(2) values for the correlation between arachidonic acid (ARA) levels in capillary whole blood with ARA levels in RBCs and plasma phospholipids (PLs) were relatively lower (R(2) = 0.31-0.41, respectively). Results indicate that the described finger stick assay represents a fast, reliable method to measure specific LC-PUFA levels.
开发了一种涉及手指采血和滤纸血斑的检测方法,以测定血液中的多不饱和脂肪酸(PUFA)水平。手指采血获得的毛细血管全血被滴在浸渍有抗氧化剂的滤纸上,风干,使用氢氧化钠和三氟化硼的甲醇溶液进行皂化和甲基化。该方法与先前描述的方法不同,因为不需要分离血浆和红细胞(RBC),不需要用薄层色谱法(TLC)分离磷脂,在酯交换反应之前不需要预先提取脂质,并且脂肪酸甲酯(FAME)方法能够使甾醇酯、游离脂肪酸和鞘磷脂甲基化。26名受试者通过手指采血和静脉穿刺提供了血样。毛细血管全血中长链多不饱和脂肪酸(LC-PUFA)的水平与静脉血中红细胞和磷脂中的水平相关(P < 0.001,R²范围为0.64至0.86)。尽管具有高度显著性(P < 0.002),但毛细血管全血中花生四烯酸(ARA)水平与红细胞和血浆磷脂(PL)中ARA水平之间相关性的R²值相对较低(分别为R² = 0.31 - 0.41)。结果表明,所描述的手指采血检测方法是一种测量特定LC-PUFA水平的快速、可靠的方法。