Analysis of sorbitol, galactitol, and myo-inositol in lens and sciatic nerve by high-performance liquid chromatography.
Accumulation of sorbitol or galactitol and depletion of myo-inositol in hyperglycemic conditions such as diabetes and galactosemia involve the activity of aldose reductase and are implicated in hyperglycemia-induced complications such as cataract and neuropathy. A high-performance liquid chromatographic method has been developed for the measurement of polyols in the lens and sciatic nerve of rats. This method comprises polyol extraction from tissues, lyophilization of extracts, derivatization of polyols by the reaction with phenylisocyanate, and HPLC of derivatives with detection at 240 nm. The time needed for each run is less than 25 min, which allows the testing of a large number of samples per day. Sensitivity is very high: as low as 0.5 nmol each of sorbitol, galactitol, and myo-inositol in lyophilized extracts of tissues can be determined. The present method offers a reliable tool to evaluate the in vivo activities of aldose reductase and its inhibitors.
Miwa
I
I
Department of Clinical Biochemistry, Faculty of Pharmacy, Meijo University, Nagoya, Japan.
Kanbara
M
M
Wakazono
H
H
Okuda
J
J
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism; D004376:Galactitol / Q000032:analysis; D005693:Galactosemias / Q000378:metabolism; D007294:Inositol / Q000032:analysis; D007908:Lens, Crystalline / Q000032:analysis*; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D012584:Sciatic Nerve / Q000032:analysis*; D013012:Sorbitol / Q000032:analysis; D013402:Sugar Alcohols / Q000032:analysis*
D013402:Sugar Alcohols; D004376:Galactitol; D007294:Inositol; D013012:Sorbitol