title: Development and validation of a high performance liquid chromatographic method for simultaneous determination of vitamins A and D3 in fluid milk products.
abstract: An HPLC method for simultaneous determination of vitamins A and D3 in fluid milk was developed and validated. Saponification and extraction conditions were studied for optimum recovery and simplicity. An RP HPLC system equipped with a C18 column and diode array detector was used for quantitation. The method was subjected to a single-laboratory validation using skim, 2% fat, and whole milk samples at concentrations of 50, 100, and 200% of the recommended fortification levels for vitamins A and D3 for Grade "A" fluid milk. The method quantitation limits for vitamins A and D3 were 0.0072 and 0.0026 μg/mL, respectively. Average recoveries between 94 and 110% and SD values ranging from 2.7 to 6.9% were obtained for both vitamins A and D3. The accuracy of the method was evaluated using a National Institute of Standards and Technology standard reference material (1849a) and proficiency test samples.
authors.affiliation: U.S. Food and Drug Administration, Center for Food Safety and Applied Nutrition, 6502 South Archer Rd, Bedford Park, IL 60501-1957.
authors.forename: Yang
authors.initials: Y
authors.lastname: Chen
authors.forename: Ravinder M
authors.initials: RM
authors.lastname: Reddy
authors.forename: Wenjing
authors.initials: W
authors.lastname: Li
authors.forename: Ramesh R
authors.initials: RR
authors.lastname: Yettlla
authors.forename: Salvador
authors.initials: S
authors.lastname: Lopez
authors.forename: Michael
authors.initials: M
authors.lastname: Woodman
mesh_terms: D000818:Animals; D002417:Cattle; D002762:Cholecalciferol / Q000737:chemistry*; D002853:Chromatography, Liquid / Q000379:methods*; D008892:Milk / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D035501:Uncertainty; D014801:Vitamin A / Q000737:chemistry*
chemical_list: D014801:Vitamin A; D002762:Cholecalciferol