Separation of Prostanoids by One-Dimensional Thin-Layer Chromatography

Abstract

We have developed a simple and rapid method for separation of 7 prostanoids and arachidonic acid by one-dimensional thin-layer chromatography. For this separation we employ commercially prepared thin-layer plates that have a preadsorbant (celite) area to which samples are applied. Due to the inert characteristics of the celite no separation occurs until the sample reaches the preadsorbant-silica gel junction. Since all the material moves with the solvent front as a sharp, narrow band to the preadsorbant (celite)-silica gel boundary, a high resolution of the separated compounds is achieved. The time required to apply one sample to the preadsorbant (celite) area is considerably less (2 min) than that required for application of a sample to silica gel (10 min). This method is suitable for the separation of major prostaglandins (PGE₂, PGF_2α), the major enzymatically formed metabolites of these prostaglandins, and the stable, nonenzymatically formed product of thromboxane A₂ (TXB₂).