Analysis of concurrent glucose consumption by the hexose monophosphate shunt, glycolysis, and the polyol pathway in the crystalline lens.
A method based on 13C-NMR spectroscopy is described, in which the metabolism of [13C2]glucose was assessed in intact rat lenses. By analysing the ratio of 13C3/(13C2 + 13C3) lactate, a quantitative measurement of the hexose monophosphate shunt (HMPS) activity could be derived. Similarly, measurements of the time-dependent increase of lactate and sorbitol permitted the determination of the activity of the Embden-Meyerhof pathway and the polyol pathway, respectively. This method offers significant advantages over other biochemical and spectroscopic methods and allows for the first time a direct correlation between the activities of the polyol pathway and the HMPS.