Batch Anion Exchange Separation and Quantification Of [14C]Hexose from [14C]Sucrose

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Abstract
Techniques quantifying 14C]photoassimilate partitioning between hexose and sucrose can identify developmental and regulatory changes in sugar metabolism of sorghum [Sorghum bicolor (L.) Moench]. The objective of this study was to develop a laboratory method for separating and quantifying [14C]hexose and [14C]sucrose in numerous plant organ extracts simultaneously. The described method combines easily with available assays for [14C]starch and [14C]structural components and for total fructose, glucose, sucrose, and starch. Treatment with hexokinase is followed by batch treatment with anion exchange resin. Hexose, as hexose phosphate, binds (>95%) to the resin, and sucrose remains unbound (>95%). Radiolabel in extracts is nearly completely recovered (99%). Determinations are reproducible (SE = 1.3%). This method was applied to identify differences in sugar metabolism between organs and among growth stages of field‐grown sorghum.

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