HPLC-PAD法による赤血球中糖およびポリオールの測定

1. 赤血球中ソルビトール

抄録

A sensitive and simple method of high-performance liquid chromatography with pulsed amperometric detection (HPLC-PAD) has been developed to determine red cell sorbitol, a potentially useful indicator of diabetic complications. Red cells were hemolyzed with deionized water and deproteinized with ultrafilter Centricon-10. Red cell sorbitol was separated with sugar pak Pb columns, eluted with deionized water at 80°C, and detected using PAD. The standard curve for sorbitol was linear up to 10.0μg/ml.The CVs for within-run precision (n=20) were 9.81% at 0.24 μg/ml and 2.65% at 4.18μg/ml. Analytical recovery was 99.3%. Red cell sorbitol values determilled by the enzymatic method (X) were higher than those obtained by the HPLC-PAD method (Y)(n=25, y=0.45x+7.80, r=0.804, X=45.04nmol/gHb, Y=28.38nmol/gHb). We analyzed red cell sorbitol from normal subjects and diabetic subjects. The sorbitol levels were 17.27±9.12 (mean±SD) nmol/gHb in the normal subjects (n=11), 31.00±12.29nmol/gHb in diabetics without complications (n=16), and 61.10±25.11nmol/gHb in diabetics with complications (n=6). The HPLC-PAD method for red cell sorbitol will be useful in assessing polyol abnormalities in diabetes mellitus.