Abstract
Objective: Sugar intake has increased vastly in the US diet during past decades and has been associated with obesity, metabolic syndrome, type 2 diabetes, cardiovascular disease, and cancer. Measuring sugar intake by FFQ is not reliable and urinary fructose and sucrose excretion as a biomarker is recommended but existing methods for their analysis are complex, expensive or unreliable (GCMS, enzymatic kits, doubly labeled water). A simple liquid chromatography mass spectrometry (LCMS) method was therefore established for urinary fructose and sucrose.Methods: This study evaluated many extraction, chemical derivatization and electrospray ionization (ESI) tandem MS variations in positive and negative mode using isotope dilution and reversed, normal or reversed reversed phase LC protocols.Results: While sucrose was measurable reliably by many LCMS protocols myriads of interferences with fructose measurements could be avoided by derivatization of fructose to its dansyl hydrazone or by selective application of organic solvents during urine extraction.Conclusions: A reliable, simple, fast and inexpensive isotope dilution tandem ESI LCMS assay for fructose and sucrose was established requiring 20 uL or less of urine.Grant Funding Source: This research was supported by National Cancer Institute grants P30 CA71789.
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