Reverse phase high-pressure liquid chromatographic separation of retinoids, including retinylphosphate and mannosylretinylphosphate

✍ Scribed by Pangala V. Bhat; Luigi M. De Luca; Marilyn L. Wind

• Publisher: Elsevier Science
• Year: 1980
• Tongue: English
• Weight: 429 KB
• Volume: 102
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(80)90346-2

✦ Synopsis

Reverse phase hplc was used to separate retinylphosphate mannosylretinylphosphate from other retinoids. These phosphorylated retinoids constituted 12 and42% of the extracted radioactivity in OEL mouse intestine at 3 and 24 h respectively after the intubation of [l-3H]retinol. A detailed procedure for the chemical synthesis of retinylphosphate is described.

Work from several laboratories (l-6) has demonstrated that mammalian tissues synthesize retinylphosphate.

Available techniques for the isolation of this compound include chromatography on columns of silicic acid, DEAE-cellulose-acetate, and thin layers of silica gel (2,3,6). However, recovery of RP* and its mannosyl derivative MRP by these techniques is poor, and, usually, losses of 60% or higher are found.

Here we describe a fast and reliable technique for the separation of RP and MRP from other retinoids, the majority of phospholipids and dolichol derivatives, in a reverse phase hplc system. We also describe an improved method to obtain synthetic retinylphosphate.

MATERIALS AND METHODS

Chemicals

Retinoic acid, retinal, retinol, and retinylacetate were a gift from Dr.