Analysis of riboflavin and riboflavin cofactor levels in plasma by high-performance liquid chromatography

• Published in: Journal of chromatography. B, Biomedical sciences and applications Vol. 739; no. 1; pp. 219 - 224
• Main Authors: Capo-chichi, Callinice D, Guéant, Jean-Louis, Feillet, François, Namour, Farès, Vidailhet, Michel
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 28.02.2000; Elsevier Science
• Subjects: Adolescent; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Child; Chromatography, High Pressure Liquid - methods; Coenzymes, vitamins; Flavin Mononucleotide - analysis; Flavin Mononucleotide - blood; Flavin nucleotides; Flavin-Adenine Dinucleotide - analysis; Flavin-Adenine Dinucleotide - blood; Fundamental and applied biological sciences. Psychology; Humans; Infant; Other biological molecules; Reference Standards; Reproducibility of Results; Riboflavin; Riboflavin - analogs & derivatives; Riboflavin - analysis; Riboflavin - blood; Riboflavin - chemistry; Riboflavin - metabolism; Vitamers; Riboflavin; Vitamers; Flavin nucleotides; Flavin derivatives; Human; Biological fluid; Analysis method; HPLC chromatography; Nucleotide; Cofactor; Quantitative analysis; Coenzyme; Blood plasma
• ISSN: 0378-4347; 1387-2273
• DOI: 10.1016/S0378-4347(99)00469-7

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 C 18 Sep-Pack cartridge after protein precipitation with 10% trichloroacetic acid, and were analyzed on a C 18 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 n M, and 36.5–157.20, 7.1–24.6 and 8.2–57.8 n M, respectively. Using GF as an internal standard improved the quantification of these B2 vitamers.