Yee H Y
Clin Chem. 1982 Nov;28(11):2241-3.
A quantitative method for lecithin and sphingomyelin is presented. In this method, which may be calibrated with a primary standard phosphate salt or pure phospholipids, 2 mL of amniotic fluid is extracted, and the solvent passed through a column of hydroxyapatite to remove all phospholipids except lecithin and sphingomyelin. After evaporation, the residue is treated with methanolic sodium hydroxide, which hydrolyzes lecithin to inorganic phosphate and a methyl ester. The unreacted sphingomyelin is removed by extraction, and the solvent evaporated. Both fractions are digested and assayed for phosphate by reduced phosphomolybdate. The total time required is 2 h. Analytical recoveries were 91-97%, and excellent agreement was found for controls. Amniotic fluid specimens were assayed to illustrate application of the proposed method. If this method is used for obtaining a lecithin/sphingomyelin ratio, a new ranges must be established, as this method is not equivalent to thin-layer chromatographic ones.
本文介绍了一种测定卵磷脂和鞘磷脂的定量方法。在该方法中，可以用一级标准磷酸盐或纯磷脂进行校准，取2 mL羊水进行萃取，使溶剂通过羟基磷灰石柱，以除去除卵磷脂和鞘磷脂之外的所有磷脂。蒸发后，残留物用甲醇氢氧化钠处理，将卵磷脂水解为无机磷酸盐和甲酯。通过萃取除去未反应的鞘磷脂，然后蒸发溶剂。将两个馏分进行消化，并通过还原磷钼酸盐测定磷酸盐含量。所需总时间为2小时。分析回收率为91 - 97%，对照结果显示出极佳的一致性。对羊水标本进行了测定，以说明所提出方法的应用。如果使用该方法来获得卵磷脂/鞘磷脂比值，则必须建立新的范围，因为该方法与薄层色谱法不等同。
