title: Indirect competitive immunoassay for detection of vitamin B₂ in foods and pharmaceuticals.
abstract: An indirect immunoassay for the determination of vitamin B2 in food samples and vitamin tablets was developed. A carbodiimide-modified active ester method was used to synthesize the immunogen for vitamin B2. The coupling ratio of vitamin B2 to carrier protein in immunogen was 19.98:1. The titer of the polyclonal antibody was 1:64000, and the antibody showed high specificity in the presence of vitamin B2 photolytic products and other B group vitamins. The immunoassay showed detection limits (LODs) of 1.07 ng/mL in PBS, 24.6 ng/g in vitamin drink, and 0.50 mg/kg in milk powder. Recovery was 99.58-110.91% in milk powder and 70.20-100.5% in vitamin drink. Vitamin B2 samples were analyzed by high-pressure liquid chromatography (HPLC) and the immunoassay, and results showed good agreement. Finally, this method was applied to detect vitamin B2 in commercial milk powder and vitamin tablets, and the detected amount correlated well with the labeled amount.
authors.affiliation: College of Pharmacy, State Key Laboratory of Medicinal Chemical Biology and Tianjin Key Laboratory of Molecular Drug Research, Nankai University , Tianjin 300071, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Yin
authors.forename: Sergei A
authors.initials: SA
authors.lastname: Eremin
authors.forename: Victor B
authors.initials: VB
authors.lastname: Rybakov
authors.forename: Taichang
authors.initials: T
authors.lastname: Zhang
authors.forename: Zhihuan
authors.initials: Z
authors.lastname: Xu
authors.forename: Linlin
authors.initials: L
authors.lastname: Ren
authors.forename: Xiaodan
authors.initials: X
authors.lastname: He
authors.forename: Meng
authors.initials: M
authors.lastname: Meng
authors.forename: Rimo
authors.initials: R
authors.lastname: Xi
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis / Q000379:methods*; D057230:Limit of Detection; D008297:Male; D008892:Milk / Q000737:chemistry; D011817:Rabbits; D012256:Riboflavin / Q000032:analysis* / Q000276:immunology; D012680:Sensitivity and Specificity; D013607:Tablets / Q000032:analysis
chemical_list: D013607:Tablets; D012256:Riboflavin