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Analysis of dipalmitoyl phosphatidylcholine in amniotic fluid by enzymatic hydrolysis and high-performance thin-layer
J G Alvarez1, B Slomovic, J Ludmir
1Department of Obstetrics and Gynecology, Beth Israel Hospital, Harvard Medical School, Boston, MA 02215, USA.
Abstract:
A novel test for the determination of dipalmitoyl phosphatidylcholine (DPPC) in amniotic fluid (AF) as free dipalmitoylglycerol (DPG), is described. Aliquots of amniotic fluid were hydrolyzed with Bacilus cereus phospholipase C, and the resulting diglycerides analyzed by AgNO3-modified high-performance thin-layer chromatography (HPTLC)-reflectance spectrodensitometry. This HPTLC system provided resolution of DPG and palmitoylpalmitoleoylglycerol (POG) from other 1,2-diglycerides and cholesterol. The turn-around analysis time for triplicate aliquots of amniotic fluid was 40 min. Recoveries ranged between 90 and 98%. In summary, this method provides a quantitative, specific, highly reproducible, and fast turn-around means of analysis of DPPC in amniotic fluid.