A new colorimetric method for determining antioxidant levels using 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS)
Takeki Hamasaki,
Taichi Kashiwagi,
Takaaki Komatsu,
Shigeru Kabayama,
Noboru Nakamichi,
Kiichiro Teruya,
Sanetaka Shirahata
Affiliations
Takeki Hamasaki
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan; Corresponding author.
Taichi Kashiwagi
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan; Department of Histology and Neuroanatomy, Tokyo Medical University, 6-1-1, Shinjuku, Shinjuku-ku, 160-8402, Japan
Takaaki Komatsu
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan; Department of Pharmacology Daiichi University of Pharmacy 22-1 Tamagawa-cho, Minami-ku, Fukuoka, 815-8511, Japan
Shigeru Kabayama
Nihon Trim Co. Ltd, 1-8-34 Oyodonaka, Kita-ku, Osaka, 531-0076, Japan
Noboru Nakamichi
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan
Kiichiro Teruya
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan
Sanetaka Shirahata
Department of Genetic Resources Technology, Faculty of Agriculture, Kyushu University, 744, Motooka, Nishi-ku, Fukuoka, 819-0395, Japan
We describe here a novel assay that determines the total a+ntioxidative activities of known antioxidants and antioxidants in beverages. The method employs the substrate 3,5-dibromo-4-nitrosobenzene sulfonate (DBNBS) that yields the colored product 3,5,3’,5’-tetrabromoazobenzene sulfate sodium salt (azo-TBBS). The amounts of azo-TBBS are measured using HPLC and then used to calculate total antioxidative capacity (TAC) values. We first show that the TAC values measured using the new DBNBS system were significantly higher compared with the control. The assay was validated through further analysis of 56 compounds, including previously characterized antioxidants. The data are consistent with published values. Here we describe in detail the application of the DBNBS method to the measurement of the TAC values of eight beverages, including wines and fruit juices. The DBNBS assay employs a readily applicable protocol that sensitively determines the levels of antioxidants in foodstuffs.- A new DBNBS-mediated antioxidant assay system is compared with standard DPPH and ORAC assays- DBNBS traps hydrogen radicals to generate a readily measured colored reduction product that quantifies antioxidant levels
