Priego Capote Feliciano, Jiménez José Ruiz, Granados José María Mata, de Castro María Dolores Luque
Department of Analytical Chemistry, University of Córdoba, Spain.
Rapid Commun Mass Spectrom. 2007;21(11):1745-54. doi: 10.1002/rcm.3014.
A method for determination of fat-soluble vitamins K(1), K(3), A, D(2), D(3) and E (as alpha- and delta-tocopherol) and metabolites 25-hydroxyvitamin D(2) and D(3) and 1,25-dihydroxyvitamin D(3) in human serum by liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) in positive mode is proposed. Highly selective identification of the target compounds in serum was confirmed by the most representative transitions from precursor ion to product ion. Quantitative MS/MS analysis was carried out by multiple reaction monitoring optimizing the most sensitive transition for each analyte in order to achieve low detection limits (from 0.012 to 0.3 ng/mL estimated with serum). The analysis was performed with 1 mL of serum, which was subjected to protein precipitation, liquid-liquid extraction to an organic phase, evaporation to dryness and reconstitution with methanol. The precision of the overall method ranged from 3.17-6.76% as intra-day variability and from 5.07-11.53% as inter-day variability. The method, validated by the standard addition method, provides complete information on the fat-soluble vitamins profile, which is of interest in clinical and metabolomics studies.
本文提出了一种采用液相色谱/电喷雾电离串联质谱法（LC/ESI-MS/MS）在正模式下测定人血清中脂溶性维生素K(1)、K(3)、A、D(2)、D(3)和E（α-生育酚和δ-生育酚）以及代谢物25-羟基维生素D(2)、D(3)和1,25-二羟基维生素D(3)的方法。通过从前体离子到产物离子的最具代表性的跃迁，证实了血清中目标化合物的高选择性鉴定。通过多反应监测对每种分析物最灵敏的跃迁进行优化，以实现低检测限（血清估计检测限为0.012至0.3 ng/mL），从而进行定量MS/MS分析。分析采用1 mL血清，先进行蛋白质沉淀，再液-液萃取至有机相，蒸发至干，最后用甲醇复溶。整个方法的精密度日内变异范围为3.17 - 6.76%，日间变异范围为5.07 - 11.53%。该方法经标准加入法验证，可提供有关脂溶性维生素谱的完整信息，这在临床和代谢组学研究中具有重要意义。
