On the suitability of refractometry for the analysis of glucose in blood-derived fluids

Refractometry is the determination of the optical refractive index of a substance or a mixture of substances. It is a very sensitive method for the detection and quantification of dissolved analytes, but it is incapable of distinguishing between different analytes. The aim of this investigation was to determine the principle suitability of refractometry for the quantification of glucose (blood sugar) in blood and various blood fluids which can readily be obtained for medical diagnosis, in particular blood plasma, blood serum, and their ultrafiltrates.

After the oral intake of freshly dissolved alpha-glucose, the in vivo blood contents of the alpha and beta anomers of glucose were found to be in an at least approximate equilibrium at all times. This observation is a prerequisite for a refractometrical determination of glucose due to the fact that both molecule forms have different refractive index increments. An assessment of the glucose content in untreated blood fluids was not possible, since no suitable relationship to the refractive index was found, most probably due to the influence of the many other substances present in blood on this parameter. However, after removal of certain macromolecules by ultrafiltration, value pairs showed a high level of correlation, providing the nominal molecular weight limit (cut-off) of the ultrafilter used possessed a maximum of 300 kDa. Besides macromolecules, the osmolality of the fluids undergoing measurement also proved to be a considerable interfering factor, particularly when values were outside the normal physiological range between 285 and 293 mmol/L.

If a clinical application of this method is to be contemplated it is imperative (1) that blood cells are separated and removed, (2) that macromolecules present in plasma or serum are removed, e.g. by ultrafiltration, and (3) that beyond the results presented the influence of all small molecules other than glucose on the overall refractive index be determined and included in the calculation of analysis results.