A sensitive assay of red blood cell sorbitol level by high performance liquid chromatography: potential for diagnostic evaluation of diabetes.
BACKGROUND
The level of sorbitol within erythrocyte or other tissue is increased during diabetic complications such as neuropathy, retinopathy, and nephropathy and is thus considered as an indicator for the diagnosis and monitoring the diabetic status. A highly sensitive method of estimation of RBC sorbitol may be very useful in determining the state of diabetes. In this study a high performance liquid chromatography method was developed to analyze RBC sorbitol level.


METHODS
Benzoylation was used for the precolumn derivatization of RBC polyol prior to analyze by HPLC using C-18 reversed phase column and UV detector.


RESULTS
The sensitivity was 5 ng/ml using standard sorbitol. The linearity was highly satisfactory up to 2.5 mug/ml which was adequate to measure RBC sorbitol in both diabetic and normal subjects. The values were significantly higher for the diabetic patient compared to normal. Within-run assay CVs were <10% and <14% for between-run.


CONCLUSIONS
This method could be useful in estimating RBC sorbitol content for the clinical evaluation of diabetic patients.
Kwang-Hyok
Son
S
Department of Cellular Biochemistry, Indian Institute of Chemical Biology, 4, Raja S. C. Mullick Road, Kolkata-700 032, India.
Ui-Nam
Pak
P
Sarkar
Chinmoy
C
Bhadra
Ranjan
R
D000328:Adult; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003924:Diabetes Mellitus, Type 2 / Q000097:blood / Q000175:diagnosis*; D004912:Erythrocytes / Q000737:chemistry*; D006801:Humans; D008875:Middle Aged; D012680:Sensitivity and Specificity; D013012:Sorbitol / Q000032:analysis* / Q000592:standards
D013012:Sorbitol