Determination of hemoglobin based on its enzymatic activity for the oxidation of o-phenylenediamine with hydrogen peroxide

Zhang, Kelly; Cai, Ruxiu; Chen, Danhua; Mao, Luyuan

doi:10.1016/s0003-2670(00)00752-2

Cited by 87 publications

(36 citation statements)

References 19 publications

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“…3. It is obvious that the peroxidase activity of Hb in Na 2 HPO 4 -NaOH buffer solution was much higher than that in Tris-HCl, NH 3 -NH 4 Cl and glycine-NaCl-NaOH buffers. So the Na 2 HPO 4 -NaOH buffer system was adopted to study the influence of different pH on the sensitivity of this method.…”

Section: Optimal Conditions For Determination Of Hbmentioning

confidence: 93%

“…A number of methods, such as spectrophotometry, [1][2][3][4] fluorimetry, [5][6][7][8] electrochemistry, 9,10 chemiluminescence 11 and high performance liquid chromatography 12 have been developed for the measurement of Hb. Among them, the most widely used method is spectrophotometric method, which mainly include the cyanomethemoglobin method (HiCN) 1 and the tetramethylbenzidine ( TMB ) method.…”

Section: Introductionmentioning

confidence: 99%

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New Fluorimetric Determination of Hemoglobin Used as a Substitute of Mimietic Peroxidase

Shi-hui

2006

Annali di Chimica

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Summary -Hemoglobin ( Hb ) could be used as a substitute of peroxidase in the catalytic oxidation of tetra-substituted amino aluminum phthalocyanine ( TAAlPc ) by H 2 O 2 . We found that the fluorescence of TAAlPc (a red-region fluorescent dye with a maximum excitation wavelength at 606 nm and a maximum emission wavelength at 673 nm) could significantly be quenched by H 2 O 2 in the presence of Hb. The value of F 0 /F ( where the relative fluorescence intensity of blank solution and that of the sample solution containing Hb were given by F 0 and F, respectively) is linearly related to the concentration of Hb. Based on this, a novel fluorimetric method was developed for the determination of Hb in aqueous solution. Under optimal conditions, Hb could be determined in the concentration range of 5× 10 -11 -1.2×10-8 mol L -1 with a detection limit of 1.5×10 -11 mol L -1 . The relative standard deviation of ten replicate measurements was 1.95% for solution containing 1×10 -9 mol L -1 Hb. The proposed method has been applied to the analysis of Hb in human blood and the results were in good agreement with those reported by a hospital laboratory. So this is a new, high sensitive and precise fluorescence quenching method to determine Hb.

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Section: Optimal Conditions For Determination Of Hbmentioning

confidence: 93%

Section: Introductionmentioning

confidence: 99%

New Fluorimetric Determination of Hemoglobin Used as a Substitute of Mimietic Peroxidase

Shi-hui

2006

Annali di Chimica

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“…Hb has oxidant activities and it has been proved that Hb can effectively act as mimetic peroxidase [5,6] . Since Hb has a quaternary structure which consists of four polypeptide subunits (two α, β protomers) and each contains a heme (iron-porphyrin) as the active center.…”

Section: Introductionmentioning

confidence: 99%

Hemoglobin-biocatalyzed synthesis of conducting molecular complex of polyaniline and lignosulfonate

Liu

Zhao

et al. 2008

J. Wuhan Univ. Technol.-Mat. Sci. Edit.

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A new biocatalyst route for the synthesis of a conducting polyaniline (PANI)/ lignosulfonate (LGS) complex was presented. Four different catalysts such as hemoglobin (Hb), 5, 10, 15, 20-tetrakis (meso-hydroxyphenyl) porphyrin, iron (II) tetrasulfophthalocyanine and ferric chloride were used to polymerize aniline in the presence of a natural polyelectrolytes template LGS. The experimental results show that Hb is an effective catalyst in this case and the synthesis is simple, and the conditions are mild in that the polymerization may be carried out in lower pH (1.0-4.0) buffered solution and optimal pH of 2.0. Varying concentrations of aniline, LGS and H 2 O 2 in feed the favorable conditions for the production of PANI were determined. UV-vis absorption, FTIR, elemental analysis, conductivity, cyclic voltammetry and thermogravimetric analyses confirm the formation of thermally stable and electroactive PANI.

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“…It contains four subunits of polypeptide, and each polypeptide chain contains a heme group that serves as the active center. Thus Hb can be a mimetic enzyme of HRP, 7,20 and the catalytic activity of Hb is much higher than hemin, β-CD-hemin and some metalloporphyrins because Hb has the natural quaternary structure. 7,20 Furthermore, Hb is very stable, and it is worth to mentioning that the price of Hb is 500-times less than that of HRP.…”

Section: Introductionmentioning

confidence: 99%

“…Thus Hb can be a mimetic enzyme of HRP, 7,20 and the catalytic activity of Hb is much higher than hemin, β-CD-hemin and some metalloporphyrins because Hb has the natural quaternary structure. 7,20 Furthermore, Hb is very stable, and it is worth to mentioning that the price of Hb is 500-times less than that of HRP. There is little doubt that Hb can be effectively substituted for HRP for H2O2 determination.…”

mentioning

confidence: 99%

Fluorescence Determination of Hydrogen Peroxide Using Hemoglobin as a Mimetic Enzyme of Peroxidase

Zhang

2001

ANAL. SCI.

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The determination of hydrogen peroxide is of great importance, ascribable to both the fact that it is a product of reactions catalyzed by a large number of oxidase enzymes, and that it is essential in food, pharmaceutical, environmental, and clinical analyses. 1,2 Many analytical methods have been reported for such measurements, including electroanalytical techniques, 3-5 spectrophotometry, 6,7 chemiluminescence, 8,9 and fluorometry.10-12 Among these methods, most are based on the enzyme (peroxidase and catalase) catalytic reaction. Enzymatic determinations of H2O2 have drawn increasing interest due to their unusual sensitivity, selectivity and simplicity. Horse radish peroxidase (HRP), in which the heme iron acts as the activity center, is the most commonly used enzyme in H2O2 detection. However, the instability and high cost of the natural enzyme has stimulated people to search for alternatives. So far, peroxidase mimics that had been applied to the determination of H2O2 have been based on two kinds of substances: one is small bio-molecules from bovine blood, such as hemin and hematin, 10,13,14 and the other is the synthesized metal complexes, such as metal-porphyrin complexes, [15][16][17] metalphthalocyanines. 18,19 However, simple, natural or synthetic metal-complexes do not show satisfactory activity and selectivity because they lack the spatial structure of the natural enzyme, which is essential for the special inclusion behavior between the enzyme and the substrate.Hemoglobin (Hb), a necessary vehicle for oxygen carriage, has the natural quaternary structure. It contains four subunits of polypeptide, and each polypeptide chain contains a heme group that serves as the active center. Thus Hb can be a mimetic enzyme of HRP, 7,20 and the catalytic activity of Hb is much higher than hemin, β-CD-hemin and some metalloporphyrins because Hb has the natural quaternary structure. 7,20 Furthermore, Hb is very stable, and it is worth to mentioning that the price of Hb is 500-times less than that of HRP. There is little doubt that Hb can be effectively substituted for HRP for H2O2 determination. However, to the best of our knowledge, the application of Hb as mimetic peroxidase to a fluorescence analysis of H2O2 has not yet been reported. In this paper, a novel fluorometry for hydrogen peroxide determination is presented, in which hemoglobin (Hb) was used as mimetic enzyme of peroxidase for fluorogenic reaction between an inexpensive substrate thiamine and H2O2. Under the catalysis of Hb, thiamin was oxidized by H2O2 to a fluorescent product, thiochrome, which exhibited strong fluorescence at 440 nm with an excitation wavelength of 370 nm. The obtained result demonstrates that Hb is a promising peroxidase mimic. The applicability of the catalytic reaction was adopted to the determination of H2O2 in rainwater. Satisfactory results were obtained. Experimental ReagentsHb (bovine) solutions were prepared by dissolving a certain amount of Hb (Sino-American Biotechnology Co., China) in water and stored in refrigerator (4...

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Determination of hemoglobin based on its enzymatic activity for the oxidation of o-phenylenediamine with hydrogen peroxide

Cited by 87 publications

References 19 publications

New Fluorimetric Determination of Hemoglobin Used as a Substitute of Mimietic Peroxidase

New Fluorimetric Determination of Hemoglobin Used as a Substitute of Mimietic Peroxidase

Hemoglobin-biocatalyzed synthesis of conducting molecular complex of polyaniline and lignosulfonate

Fluorescence Determination of Hydrogen Peroxide Using Hemoglobin as a Mimetic Enzyme of Peroxidase

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