Enhanced UV Detection of Sugar Phosphates by Addition of a Metal Complex to the HPLC Mobile Phase

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Abstract
This paper demonstrates the use of a mobile phase additive, Eu(DPM)3, to enhance the UV detection of phosphorylated sugars. The phosphorylated sugars were separated on a strong anion exchange HPLC column using a formate buffer containing 2 × 10 −5M Eu(DPM)3. The eluent was monitored by UV detection at 254 nm in series with refractive index detection. Eleven commercially available phosphorylated sugars were analyzed under the conditions found to be optimal for both UV detection and chromatographic behavior. Retention data and k∲ values are given. Spectrophotometric studies of glucose-6-phosphate, Eu(DPM)3, and glucose-6-phosphate complexes with Eu(DPM)3 were performed, and changes in absorbance were found that enhance the UV detection of glucose-6-phosphate.