Identification and determination of fat-soluble vitamins and metabolites in human serum by liquid chromatography/triple quadrupole mass spectrometry with multiple reaction monitoring.
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.
Priego Capote
Feliciano
F
Department of Analytical Chemistry, University of Córdoba, Spain. q72prcaf@uco.es
Jiménez
José Ruiz
JR
Granados
José María Mata
JM
de Castro
María Dolores Luque
MD
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D005223:Fats / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D012995:Solubility; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D014815:Vitamins / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification
D005223:Fats; D014815:Vitamins