title: Dipstick based immunochemiluminescence biosensor for the analysis of vitamin B12 in energy drinks: a novel approach.
abstract: In this article, we describe a dipstick based immunochemiluminescence (immuno-CL) biosensor for the detection of vitamin B(12) in energy drinks. The method is a direct competitive type format involving the immobilization of vitamin B(12) antibody on nitrocellulose membrane (NC) followed by treatment with vitamin B(12) and vitamin B(12)-alkaline phosphatase conjugate to facilitate the competitive binding. The dipstick was further treated with substrate disodium 2-chloro-5-(4-methoxyspiro {1,2-dioxetane-3,2¢-(5¢-chloro)tricyclo[3.3.1.13,7]decan}-4-yl)-1-phenyl phosphate (CDP-Star) to generate chemiluminescence (CL). The number of photons generated was inversely proportional to the vitamin B(12) concentration. After systematic optimization, the limit of detection was 1 ng mL(-1). The coefficient of variation was below 0.2% for both intra- and inter-assay precision. Vitamin B(12) was extracted from energy drinks with recovery ranged from 90 to 99.4%. Two different energy drinks samples were analyzed, and a good correlation was observed when the data were compared with a reference enzyme linked immuno sorbent assay (ELISA) method. The developed method is suitable for an accurate, sensitive, and high-throughput screening of vitamin B(12) in energy drinks samples. The dipstick technique based on immuno-CL is suitable for the detection of several analyte in food and environmental samples.
authors.affiliation: Fermentation Technology and Bioengineering Department, Central Food Technological Research Institute, Mysore 570020, Karnataka, India.
authors.forename: L S
authors.initials: LS
authors.lastname: Selvakumar
authors.forename: M S
authors.initials: MS
authors.lastname: Thakur
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism; D055910:Antibodies, Immobilized / Q000276:immunology; D015374:Biosensing Techniques; D003101:Collodion / Q000737:chemistry; D061215:Energy Drinks / Q000032:analysis*; D006241:Haptens / Q000737:chemistry / Q000276:immunology; D007118:Immunoassay; D008163:Luminescent Measurements; D014805:Vitamin B 12 / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification
chemical_list: D055910:Antibodies, Immobilized; D006241:Haptens; D003101:Collodion; D000469:Alkaline Phosphatase; D014805:Vitamin B 12