Abstract：
Disturbances in the diurnal pattern are associated with several clinical and psychological conditions, including depression and fatigue. Salivary sampling for melatonin, cortisol and cortisone provides a non-invasive method for frequent sampling and obtaining biochemical insight into the diurnal pattern of individuals. Therefore, a new liquid chromatography-tandem mass spectrometry-based method for the measurement of salivary melatonin, cortisol and cortisone was developed and validated. The method required 250 μl saliva, used isotope dilution methodology and was based on a liquid-liquid extraction for sample preparation, reversed-phase chromatography and multiple reaction monitoring on a mass spectrometer for quantitation. The lower limits of quantification obtained were 0.010 nmol/L for melatonin, 0.5 nmol/L for cortisol and 1.00 nmol/L for cortisone and the limits of detection were 0.003 nmol/L, 0.15 nmol/L and 0.1 nmol/L respectively. The method imprecision was ≤14% for all measurands, and the method comparison showed highly comparable results with high correlation coefficients (all ≥0.964). Potential interference of cortisol and cortisone by prednisolone was observed and could be detected by chromatogram review. Typical diurnal patterns for melatonin, cortisol and cortisone were observed in the saliva of 20 cancer survivors who collected saliva throughout the day.
摘要：
昼夜模式的紊乱与几种临床和心理状况有关，包括抑郁和疲劳。唾液取样检测褪黑激素,皮质醇和可的松提供了一种非侵入性方法，用于频繁采样并获得对个体昼夜模式的生化见解。因此,一种新的基于液相色谱-串联质谱的唾液褪黑素测定方法，皮质醇和可的松被开发和验证。该方法需要250μl唾液，使用同位素稀释方法，并基于液-液萃取进行样品制备，在质谱仪上进行反相色谱和多反应监测以进行定量。褪黑素的定量下限为0.010nmol/L，皮质醇为0.5nmol/L，可的松为1.00nmol/L，检出限为0.003nmol/L，0.15nmol/L和0.1nmol/L。所有被测量的方法不精密度≤14%，方法比较显示结果具有高度可比性，相关系数高（均≥0.964）。观察到泼尼松龙对皮质醇和可的松的潜在干扰，并通过色谱图检查可以检测到。褪黑激素的典型昼夜模式，在全天收集唾液的20名癌症幸存者的唾液中观察到皮质醇和可的松。
