Objectives: We sought to develop an assay to measure circulating fructose concentrations in pancreatic cancer patients.
Methods: Using fructose dehydrogenase-catalyzed conversion of d-fructose to 5-ketofructose, followed by quantitation of MTT [3-(4,5-dimethylthiaze-syl)-2,5-diphenyltetrazolium bromide] formazan production by direct spectrophotometry, an assay to measure serum fructose concentration was developed, and assay sensitivity and specificity were tested. Validity of the assay was confirmed by gas chromatography-mass spectroscopy, and the assay was tested in healthy subjects and pancreatic cancer patients.
Results: The assay was highly specific, exhibiting no cross-reactivity with other sugars. Mean serum fructose concentration in fasting healthy volunteers was 1.9+/-0.4 mM and after ingestion of a fructose and glucose-containing drink rose to 16.3+/-1.2 mM at 15 minutes and peaked at 30 minutes when serum fructose was 17.2+/-1.1 mM. Mean fasting serum fructose level was significantly higher at 5.7+/-2.5 mM in patients with pancreatic cancer than those with no pancreatic cancer. The fructose dehydrogenase-based enzymatic assay correlated highly with gas chromatography-mass spectroscopic analysis of serum fructose with a correlation coefficient of 0.94.
Conclusion: This spectrophotometric assay for fructose is easy to perform and well suited to determination of serum fructose. Measurement of serum fructose concentration may provide insight into the relationship between refined fructose intake and diseases including pancreatic cancer.