Micro-quantitation of lens myo-inositol by anion exchange chromatography

Abstract

A method capable of the determination of pmole quantities of myo-inositol in mgr amounts of tissue by anion exchange chromatography is detailed for use in lens and potentially other tissues. Samples were rendered protein-free through ZnS0₄/Ba(0H)₂ precipitation, lyophilized and reconstituted in water just prior to analysis. An aliquot of sample was injected onto an anion exchange column and eluted with a 0.045 M sodium hydroxide mobile phase. Each analysis requires 30 minutes to complete. Average recovery of myo-inositol added to lens sample prior to injection was 100.6%. The coefficient of variation for repeated sample injections was 2.9%. Rabbit lens averaged 11.4 μmol/gr wet weight with epithelium containing 8.5 μmol/gr wet weight while human lens contained 20.1 μmol/gr wet weight and human epithelial cells had 17.5 nmol/mgr protein.