Preanalytical factors and reference intervals for serum hepcidin LC–MS/MS method

BackgroundHepcidin is a potential biomarker for anemia of chronic diseases and disorders of iron metabolism. Thus, data of preanalytical factors, reference values and method characteristics are critical for clinical use of hepcidin determination.MethodsWe studied the effects of sample storage, sampling device and a meal on hepcidin levels using a liquid-chromatography tandem mass spectrometric (LC–MS/MS) method for serum hepcidin. Age- and gender-dependent reference values were determined using serum samples from healthy volunteers (n = 231). The results were also compared with those obtained by a commercial competitive ELISA for hepcidin.ResultsIn serum samples, hepcidin is stable for one day at room temperature, six days at + 4 °C and at least 42 days at − 20 °C. Breakfast or type of sampling device does not affect hepcidin concentration. Significantly lower hepcidin concentrations were observed in women ≤ 50 than > 50 years of age or in men (p < 0.0001 for both). Reference values for females aged 18–50 years were 0.4–9.2 nmol/L, for those > 50 years 0.7–16.8 nmol/L and for males ≥ 18 years 1.1–15.6 nmol/L. Comparison with a competitive ELISA showed poor correlation.ConclusionsFasting before sampling and type of blood collection were not critical. Samples can be transported to laboratory at room temperature if they arrive within a day. Significantly lower concentrations of serum hepcidin were observed in menstruating than in post-menopausal women and in men.

► Nutritional status or type of blood collection does not affect hepcidin concentration. ► Serum samples are stable for one day at room temperature. ► For longer storage times samples must be refrigerated or frozen. ► Serum hepcidin is stable for at least five freeze–thaw cycles. ► Serum hepcidin is lower in menstruating than in post-menopausal women and in men.