Micro-quantitation of lens myo-inositol by anion exchange chromatography.
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 ZnSO4/Ba(OH)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 myoinositol 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 mumol/gr wet weight with epithelium containing 8.5 mumol/gr wet weight while human lens contained 20.1 mumol/gr wet weight and human epithelial cells had 17.5 nmol/mgr protein.
Dickerson
J E
JE
Cataract Research, Alcon Laboratories, Inc., Fort Worth, TX 76134.
Lou
M F
MF
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004848:Epithelium / Q000032:analysis; D006801:Humans; D007294:Inositol / Q000032:analysis*; D007904:Lens Cortex, Crystalline / Q000032:analysis; D007907:Lens Nucleus, Crystalline / Q000032:analysis; D007908:Lens, Crystalline / Q000032:analysis*; D051379:Mice; D011817:Rabbits; D051381:Rats; D011919:Rats, Inbred Strains; D012756:Sheep; D013045:Species Specificity; D013552:Swine
D007294:Inositol