A novel ion pair reversed phase ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC–MS/MS) method for simultaneous determination of the stress hormones adrenaline, noradrenaline and corticosterone in rodent blood was developed and fully validated. Separations were performed on an Acquity HSS T3 column (2.1 mm i.d. × 100 mm, 1.8 μm) with gradient elution and a runtime of 5.5 min. The retention of adrenaline and noradrenaline was substantially increased by employing the ion pair reagent heptafluorobutyric acid (HFBA). Ion pair reagents are usually added to the mobile phase only, but we demonstrate for the first time that including HFBA to the sample reconstitution solvent as well, has a major impact on the chromatography of these compounds. The stability of adrenaline and corticosterone in rodent blood was investigated using the surrogate analytes adrenaline-d₃ and corticosterone-d₈. The applicability of the described method was demonstrated by measuring the concentration of stress hormones in rodent blood samples.

建立了同时测定啮齿动物血液中应激激素肾上腺素，去甲肾上腺素和皮质酮的新型离子对反相超高效液相色谱-串联质谱（UHPLC-MS / MS）方法。分离在Acquity HSS T3色谱柱（内径2.1 mm×100 mm，1.8μm）上进行，梯度洗脱，运行时间为5.5分钟。通过使用离子对试剂七氟丁酸（HFBA），肾上腺素和去甲肾上腺素的保留量显着增加。离子对试剂通常仅添加到流动相中，但是我们首次证明了将HFBA也包含在样品重构溶剂中对这些化合物的色谱法具有重大影响。₃和皮质酮-d ₈。通过测量啮齿动物血样中应激激素的浓度证明了上述方法的适用性。