title: Development of ultrasensitive direct chemiluminescent enzyme immunoassay for determination of aflatoxin B1 in food products.
abstract: A direct competitive chemiluminescent enzyme-linked immunosorbent assay (CL-ELISA) for determination of aflatoxin B1 (AFB1) was developed. To improve the assay sensitivity, a mixture of 3-(10'-phenothiazinyl)-propane-1-sulfonate and 4-morpholinopyridine previously optimized by a factorial design was used as enhancer of horseradish peroxidase-induced chemiluminescence. Varying the concentrations of the coating anti-AFB1 antibody and conjugate of AFB1 and horseradish peroxidase the conditions of the chemiluminescent assay were optimized. The values of the detection limit value and dynamic working range of CL-ELISA of AFB1 were 0.0015 ng mL(-1) and 0.003-0.03 ng mL(-1), respectively. It was shown that a dilution of rice and mung beans extracts in 5 and 10 times, respectively, prevented a matrix effect of the food products in CL-ELISA. The recovery values from the spiked samples of rice and mung beans were in the range of 90-104% and 102-117%, respectively. Studying 8 rice and 8 mung beans samples purchased in commercial stores the developed CL-ELISA allowed to find 3 samples (1 rice and 2 mung beans) containing AFB1, the content of AFB1 in one sample being higher than the maximum acceptable level established in the European Community.
authors.affiliation: School of Biomedical Sciences, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Feng-Yih
authors.initials: FY
authors.lastname: Yu
authors.forename: Anastasia V
authors.initials: AV
authors.lastname: Gribas
authors.forename: Marina M
authors.initials: MM
authors.lastname: Vdovenko
authors.forename: Ivan Yu
authors.initials: IY
authors.lastname: Sakharov
mesh_terms: D016604:Aflatoxin B1 / Q000032:analysis*; D017738:Alkanesulfonic Acids / Q000737:chemistry; D055910:Antibodies, Immobilized / Q000737:chemistry; D001230:Aspergillus / Q000737:chemistry / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007887:Fabaceae / Q000382:microbiology; D005504:Food Analysis / Q000379:methods*; D005516:Food Microbiology / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D008163:Luminescent Measurements / Q000379:methods*; D012275:Oryza / Q000382:microbiology; D010407:Penicillium / Q000737:chemistry / Q000302:isolation & purification; D011725:Pyridines / Q000737:chemistry; D013843:Thiazines / Q000737:chemistry
chemical_list: C543492:3-(10'-Phenothiazinyl)propane-1-sulfonate; D017738:Alkanesulfonic Acids; D055910:Antibodies, Immobilized; D011725:Pyridines; D013843:Thiazines; D016604:Aflatoxin B1