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Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.

Yang L, Jin M, Du P, Chen G, Zhang C, Wang J, Jin F, Shao H, She Y, Wang S, Zheng L, Wang J - PLoS ONE (2015)

Bottom Line: 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.When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice.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.

View Article: PubMed Central - PubMed

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.

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.

No MeSH data available.


Related in: MedlinePlus

Optimization of concentration of ionic and pH value (a), H2O2 (b), Luminol (c).
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pone.0131193.g005: Optimization of concentration of ionic and pH value (a), H2O2 (b), Luminol (c).

Mentions: In the 4-IMP enhancing system, the system luminescence will be highest when the salt concentration is under 0.01M; as the pH value rises, the luminescence strength will increase. When the pH value reaches 9.0 it tends to smooth out (please see Fig 5(a)). Therefore, Tris-HCI buffer fluid (0.01M, pH9.0) provides the best chemiluminescence reaction conditions, and the enhancing system of 4-IOP and HIOP will show the same result as 4-IMP; it will present the maximum luminescence value under the condition of Tris-HCI buffer fluid (0.01M, pH9.0). Hence, the optimized Tris-HCI buffer condition for the three enhancer system is 0.01M as the salt concentration and 9.0 as the pH value.


Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.

Yang L, Jin M, Du P, Chen G, Zhang C, Wang J, Jin F, Shao H, She Y, Wang S, Zheng L, Wang J - PLoS ONE (2015)

Optimization of concentration of ionic and pH value (a), H2O2 (b), Luminol (c).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4495922&req=5

pone.0131193.g005: Optimization of concentration of ionic and pH value (a), H2O2 (b), Luminol (c).
Mentions: In the 4-IMP enhancing system, the system luminescence will be highest when the salt concentration is under 0.01M; as the pH value rises, the luminescence strength will increase. When the pH value reaches 9.0 it tends to smooth out (please see Fig 5(a)). Therefore, Tris-HCI buffer fluid (0.01M, pH9.0) provides the best chemiluminescence reaction conditions, and the enhancing system of 4-IOP and HIOP will show the same result as 4-IMP; it will present the maximum luminescence value under the condition of Tris-HCI buffer fluid (0.01M, pH9.0). Hence, the optimized Tris-HCI buffer condition for the three enhancer system is 0.01M as the salt concentration and 9.0 as the pH value.

Bottom Line: 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.When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice.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.

View Article: PubMed Central - PubMed

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.

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.

No MeSH data available.


Related in: MedlinePlus