Clinical laboratory determination of phosphatidylglycerol: one- and two-dimensional chromatography compared

Reportedly, determination of several phospholipids in amniotic fluid, including phosphatidylglycerol and phosphatidylinositol, in addition to lecithin and sphingomyelin improves prediction of neonatal pulmonary function. In this study, we evaluated a two-dimensional technique for separating and measuring these phospholipids and compared it with a simpler one-dimensional procedure. The two-dimensional technique was adapted to readily available commercial plates, and a preheating step was introduced to avoid shattering of the plates during charring. The Rf values, reproducibility of each technique, and the correlation between them were examined. Even though the one-dimensional technique is faster and less expensive, we recommend the two-dimensional method for clinical use because of better precision (CV for phosphatidylglycerol 15% vs 21%) and clearer results when relatively little phosphatidylglycerol is present. The one-dimensional procedure is unreliable when blood or meconium are present. In addition, interfering compounds co-migrate with phosphatidylglycerol, phosphatidylethanolamine, and phosphatidylserine in the one-dimensional technique. Before any one-dimensional lipid separation is adopted for clinical use, it should be critically compared to the two-dimensional procedure.