Quantification of fat-soluble vitamins in human breast milk by liquid chromatography-tandem mass spectrometry.
Sensitive quantification method for fat-soluble vitamins in human breast milk by liquid chromatography-tandem mass spectrometry was developed. Vitamins A, D and E were extracted from 10.0 mL of breast milk after saponifying by basic condition. Vitamin K derivatives were extracted from 3.0 mL of breast milk after lipase treatment. The corresponding stable isotope-labeled compounds were used as internal standards. For the determination of vitamin D compounds, derivatization with a Cookson-type reagent was performed. All fat-soluble vitamins were determined by liquid chromatography-tandem mass spectrometry in the positive ion mode. The detection limits of all analytes were 1-250 pg per 50 microL. The recoveries of fat-soluble vitamins were 91-105%. Inter-assay CV values of each vitamin were 1.9-11.9%. The mean concentrations of retinol, vitamin D3, 25-hydroxyvitamin D3, alpha-tocopherol, phylloquinone and menaquinone-4 were 0.455 microg/mL, 0.088 ng/mL, 0.081 ng/mL, 5.087 microg/mL, 3.771 ng/mL, and 1.795 ng/mL, respectively (n=82). This method makes possible to determine fat-soluble vitamins with a wide range of polarities in human breast milk. The assay may be useful for large-scale studies.
Kamao
Maya
M
Department of Hygienic Sciences, Kobe Pharmaceutical University, 4-19-1 Motoyamakita-machi, Higasihnada-ku, Kobe 658-8558, Japan.
Tsugawa
Naoko
N
Suhara
Yoshitomo
Y
Wada
Akimori
A
Mori
Toshiyuki
T
Murata
Kazuo
K
Nishino
Riichiro
R
Ukita
Tetsuya
T
Uenishi
Kazuhiro
K
Tanaka
Kiyoshi
K
Okano
Toshio
T
D002853:Chromatography, Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008895:Milk, Human / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014801:Vitamin A / Q000032:analysis*; D014807:Vitamin D / Q000032:analysis*; D014810:Vitamin E / Q000032:analysis*; D014812:Vitamin K / Q000032:analysis
D014801:Vitamin A; D014812:Vitamin K; D014807:Vitamin D; D014810:Vitamin E