31P NMR of tissue phospholipids: a comparison of three tissue pre-treatment procedures.

Abstract

Extracted tissue phospholipid 31P NMR profiles, obtained from individual porcine lenses subjected to two preservation procedures (acetone desiccation and freeze-drying) and a perchloric acid-extraction procedure, were compared to those from freshly excised lens specimens. Each profile yielded quantitative data on 12 lens phospholipids: PC, LPC, PC plas, PE, LPE, PE plas, PS, SPH, PI, LPI, PG, and CL. A specimen group size of at least 9 lenses was required for secure statistical inter-group comparisons by the Scheffé procedure, due to specimen 31P NMR profile variability, interpreted as arising from specimen biological variability. The phospholipid profiles of lenses preserved by acetone desiccation were essentially identical to those from the freshly excised control lenses. Freeze-dried lens profiles differed significantly in four components, while profiles from perchloric acid-extracted lenses differed in six. It is concluded that specimen preservation by acetone disiccation is a useful method for preserving tissue phospholipids for subsequent 31P NMR profile analysis, while freeze-drying is not. Lipid extraction following a tissue acid extraction is also of little or no value in the determination of tissue phospholipid profiles.