title: Rapid sample preparation procedure for determination of retinol and α-tocopherol in human breast milk.
abstract: The liposoluble vitamins (retinol and α-tocopherol) concentration in human breast milk is of a cardinal knowledge especially for nutrition of prematurely born. It enables the feeding optimization of these important micronutrients for preterm infants. The novel rapid liquid-liquid extraction procedure for human breast milk investigation was developed and validated according to FDA guidelines. The recovery of retinol was 82-90% measured at three concentration levels 1.0, 2.5 and 5.0 μmol/L, for α-tocopherol 92-109% at concentration levels 2.5, 5.0 and 10.0 μmol/L. The repeatability of extraction procedure expressed as relative standard deviation was 3.26% for retinol and 4.79% for α-tocopherol. Developed extraction procedure was applied on 120 human breast milk samples. The separation of vitamins was completed using advantages of a monolithic column which accomplished demands of acceleration made by modern bio-analytical HPLC methodology. The analytes of interest were detected by diode-array detector at wavelengths 325 nm for retinol and 290 nm for α-tocopherol.
authors.affiliation: Department of Analytical Chemistry, Faculty of Pharmacy, Charles University, Heyrovského 1203, 500 05 Hradec Králové, Czech Republic.
authors.forename: Markéta
authors.initials: M
authors.lastname: Kašparová
authors.forename: Jiří
authors.initials: J
authors.lastname: Plíšek
authors.forename: Dagmar
authors.initials: D
authors.lastname: Solichová
authors.forename: Lenka
authors.initials: L
authors.lastname: Krčmová
authors.forename: Barbora
authors.initials: B
authors.lastname: Kučerová
authors.forename: Miloslav
authors.initials: M
authors.lastname: Hronek
authors.forename: Petr
authors.initials: P
authors.lastname: Solich
mesh_terms: D000328:Adult; D053000:Analytic Sample Preparation Methods / Q000379:methods*; D005260:Female; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008894:Milk Proteins / Q000737:chemistry / Q000302:isolation & purification; D008895:Milk, Human / Q000737:chemistry*; D015203:Reproducibility of Results; D013997:Time Factors; D014801:Vitamin A / Q000032:analysis* / Q000302:isolation & purification*; D024502:alpha-Tocopherol / Q000032:analysis* / Q000302:isolation & purification*
chemical_list: D007202:Indicators and Reagents; D008894:Milk Proteins; D014801:Vitamin A; D024502:alpha-Tocopherol