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Isotope mass balance was then applied to calculate the 18O/16O of O-5 and O-6 as a whole and O-6, respectively. Sampling at different stages of substrate conversion to product and applying a Rayleigh-type fractionation model were employed, when quantitative conversion of substrate was unachievable to calculate the δ18O of the converted substrate. Quantitative conversion of glucose with phenylhydrazine to phenylglucosazone also allowed for the calculation of δ18O2 by applying isotope mass balance between the two. A C4 starch-derived gluco
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