title: Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.
abstract: A luminol-H2O2-HRP chemiluminescence system with high relative luminescent intensity (RLU) and long stabilization time was investigated. First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-yl)phenol (4-IMP), 4-iodophenol (4-IOP), 4-bromophenol (4-BOP) and 4-hydroxy-4'-iodobiphenyl (HIOP) had the best performance. Based on the experiment, the four enhancers were dissolved in acetone, acetonitrile, methanol, and dimethylformamide (DMF) with various concentrations, the results indicated that 4-IMP, 4-IOP, 4-BOP and HIOP dissolved in DMF with the concentrations of 0.2%, 3.2%, 1.6% and 3.2% could get the highest RLU values. Subsequently, the influences of pH, ionic strength, HRP, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol on the stabilization of the luminol-H2O2-HRP chemiluminescence system were studied, and we found that pH value, ionic strength, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol have little influence on luminescent stabilization, while HRP has a great influence. In different ranges of HRP concentration, different enhancers should be selected. When the concentration is within the range of 0~6 ng/mL, 4-IMP should be selected. When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice. And when the concentration is within the range of 25~80 ng/mL, HIOP should be selected as the enhancer. Finally, the three well-performing chemiluminescent enhanced solutions (CESs) have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP) in their utilized HRP concentration ranges.
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Yang
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Maojun
authors.initials: M
authors.lastname: Jin
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Pengfei
authors.initials: P
authors.lastname: Du
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Ge
authors.initials: G
authors.lastname: Chen
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Jin
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Shao
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: She
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Wang
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Lufei
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Key Laboratory for Agro-Products Quality and Safety, Institute of Quality Standards & Testing Technology for Agro-Products, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
mesh_terms: D000096:Acetone / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D001713:Biphenyl Compounds / Q000737:chemistry; D002021:Buffers; D004126:Dimethylformamide / Q000737:chemistry; D006735:Horseradish Peroxidase / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007093:Imidazoles / Q000737:chemistry; D007462:Iodobenzenes / Q000737:chemistry; D007700:Kinetics; D049449:Luminescence; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008165:Luminol / Q000737:chemistry*; D000432:Methanol / Q000737:chemistry; D009930:Organic Chemicals / Q000737:chemistry; D010636:Phenols / Q000737:chemistry; D010880:Piperidines / Q000737:chemistry; D012997:Solvents / Q000737:chemistry
chemical_list: C000606747:4-hydroxy-4'-iodobiphenyl; D000097:Acetonitriles; D001713:Biphenyl Compounds; D002021:Buffers; D007093:Imidazoles; D007462:Iodobenzenes; D009930:Organic Chemicals; D010636:Phenols; D010880:Piperidines; D012997:Solvents; D000096:Acetone; C085755:4-(1H-imidazol-4-ylmethyl)piperidine; C050181:4-iodophenol; D008165:Luminol; D004126:Dimethylformamide; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; C032037:4-bromophenol; D000432:Methanol; C032159:acetonitrile