Automated Carbohydrate Analysis of Physiologic Fluids

Automated carbohydrate analysis can be useful clinically in the research laboratory as an aid in understanding the fundamental role of carbohydrates in metabolism, including their pathologic significance. An analytic system being developed at our laboratory utilizes an automated carbohydrate analyzer to chromatograph borated physiologic fluids while using strongly basic anion-exchange resin. The eluted carbohydrates are detected by sulfuric acid-phenol colorimetry. The carbohydrate analyzer used in this experimental work should become a useful tool in the clinical laboratory. Normal and diabetic human urine and blood serum have been chromatographed and significant differences established. As many as 38 peaks have been observed in the complex urine chromatograms. Using cochromatographic technics, 14 peaks have been tentatively identified as sucrose, raffinose, N-acetylglucosamine, maltose, lactose, ribose, fructose, arabinose, fucose, galactose, xylose, mannoheptulose, glucose, and glucose-1-phosphate. All except fucose have been quantified. Blood serum chromatograms consist of a major glucose peak and several smaller peaks indicating traces of other sugars. Melibiose has been used as an internal standard in chromatograms to determine recovery, resolution, and reproducibility. With the present technic, the lower limit of detectable sugar in physiologic fluids is in the 1-µg. range.