title: Analysis of riboflavin and riboflavin cofactor levels in plasma by high-performance liquid chromatography.
abstract: We describe an assay which determines simultaneously riboflavin (RF), flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) in plasma, using galactoflavin (GF) as an internal standard. The flavins were extracted on a C18 Sep-Pack cartridge after protein precipitation with 10% trichloroacetic acid, and were analyzed on a C18 RP-HPLC with 85% phosphate-magnesium acetate buffer (pH 3.4) and 15% acetonitrile. FAD, FMN, GF and RF extraction recoveries were 101.0-5.6, 97.0-6.5, 97.0-2.0 and 95.0-4.1%, and reproducibilities were 5.9, 6.8, 2.1 and 4.3%, respectively. FAD, FMN and RF values in infant and adolescent plasma were in the range 53.5-108.2, 9.0-25.1 and 12.7-53.4 nM, and 36.5-157.20, 7.1-24.6 and 8.2-57.8 nM, respectively. Using GF as an internal standard improved the quantification of these B2 vitamers.
authors.affiliation: Laboratoire de Pathologie Cellulaire et Moléculaire en Nutrition, EP CNRS 616, Faculté de Médecine de Nancy, Vandoeuvre-Lès-Nancy, France.
authors.forename: C D
authors.initials: CD
authors.lastname: Capo-chichi
authors.forename: J L
authors.initials: JL
authors.lastname: Guéant
authors.forename: F
authors.initials: F
authors.lastname: Feillet
authors.forename: F
authors.initials: F
authors.lastname: Namour
authors.forename: M
authors.initials: M
authors.lastname: Vidailhet
mesh_terms: D000293:Adolescent; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005486:Flavin Mononucleotide / Q000032:analysis / Q000097:blood*; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis / Q000097:blood*; D006801:Humans; D007223:Infant; D012015:Reference Standards; D015203:Reproducibility of Results; D012256:Riboflavin / Q000031:analogs & derivatives / Q000032:analysis / Q000097:blood* / Q000737:chemistry / Q000378:metabolism
chemical_list: D005182:Flavin-Adenine Dinucleotide; D005486:Flavin Mononucleotide; D012256:Riboflavin; C007811:galactoflavin