Short-term stability of free metanephrines in plasma and whole blood

Background Analysis of plasma metanephrine (MN) and normetanephrine (NMN) with liquid chromatography tandem mass spectrometry (LC-MS/MS) is the gold standard for the screening of pheochromocytomas and paragangliomas (PPGLs). As scarce information is available on the stability of MNs in diagnostic samples, this study was aimed at analyzing the short-term stability of plasma free MNs in whole blood and plasma, using LC-MS/MS. Methods The stability of plasma MNs was evaluated after sample collection at 1, 2 and 3 h in whole blood, and at 2, 4 and 6 h in centrifuged samples. Both studies were performed while maintaining the samples at room temperature (RT) and at 4 °C. The ClinMass Complete Kit (Recipe, Munchen, Germany) was used for measuring MNs with LC-MS/MS (Nexera X2 UHPLC-4500MD Sciex). Differences from the baseline (T0) were assessed using repeated measures one-way ANOVA, Students' paired t-test and a comparison of the mean percentage changes with the total change limit (TCL). Results Statistically significant differences from T0 were found for both MNs (p < 0.001) in whole blood stored at RT, and for NMN (p = 0.028) but not MN (p = 0.220) at 4 °C. The mean difference exceeded the TCL after 1 h and 3 h at RT for MN, and after 1 h at RT for NMN. Statistically significant differences from T0 were only observed in the plasma samples for NMN at RT (p = 0.012), but the variation was within the TCL. Conclusions MN and NMN displayed different patterns of stability before and after centrifugation. Even short-time storage at RT in whole blood should hence be avoided.