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Catalytic Scattering Spectral Assay For α-amylase Activity, H2O2 And Glucose

Posted on:2009-01-10Degree:MasterType:Thesis
Country:ChinaCandidate:J MaFull Text:PDF
GTID:2121360245459562Subject:Analytical Chemistry
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Part I IntroductionSummarized the application of resonance scattering technology in analytical chemistry. The application of resonance scattering technology in analytical chemistry in resent years was summarized. The analytical progress ofα-amylase, H2O2 and glucose were introduced. Two hundred and forty-three references were quoted.Part II Resonance scattering spectra of starch and its application to assay ofα-amylase activityIn presence of 0.8mg/mL polyacrylamide(PAM), starch forms stable microparticles that the average diameter is about 317nm. It exhibits a strong resonance scattering peak at 423nm. In pH=5.3 phosphate buffer solution at 50℃, starch microparticles is catalyzed and hydrolysised into small molecules byα-amylase, which leads the resonance scattering intensity at 423nm to decrease greatly. Under proper experimental conditions, the decreased resonance scattering intensity is well linear to the concentration ofα-amylase in the range of 0.00835~0.501μg/ml (or 0.00138~0.0827U/ml), and the regression equation and the correlation coefficient are△I = 155C + 13 and 0.9857, respectively. The method is used to the determination ofα-amylase activity in human saliva, with satisfactory results.Part III HRP catalytic resonance scattering spectral determination of trace hydrogen peroxide using cationic Surfactant association particleIn acetic acid buffer solution, horseradish peroxidase (HRP) may catalyze H2O2 oxidation of excess I- to form I3-, the I3- combined with cationic surfactant (CS), such as tetradecyl dimethylbenzyl ammonium chloride (TDMAB), dodecyl dimethylbenzyl ammonium chloride(DDAC), cetyltrimethyl ammonium bromide (CTAB), dimethyl benzyl ammonium chloride(ODAC), cetylpyridinium chloride (CPCM), chlorinated dodecyl pyridine(DPC) and terabutyl ammonium iodide(TBAB) to produce association microparticles TDMBA-I3,D DAC-I3,CTAB-I3,ODC-I3,CPCM-I3,DPC-I3 and TBAB-I3, which exhibit the strongest resonance scattering peak at 460 nm, the hydrogen peroxide concentration in the range of 0.17~172.8,0.432~172.8,1.728~259.2,1.728~86.4,1.728~216,0.864~259.2 and 0.864~86.4×10-7mol?L-1, was linear to the intensity at 460 nm,with a detection limit of 0.086,0.34,1.72,0.46,0.36,0.64 and 0.44×10-7mol?L-1 respectively. The method is used to the determination of hydrogen peroxide in real samples, with satisfactory results.Part IV A highly sensitive and selective resonance scattering spectral probe for glucose based on GOD and HRP catalytic effectsIn acetic acid buffer solution, glucose (G) can be oxidized to gluconic acid and H2O2 in the presence of glucose oxidase (GOD). The H2O2 oxidized excess I- catalytically to form I3- in the presence of horseradish peroxidase (HRP). The I3- combined with cationic surfactant (CS) such as tetradecyl dimethylbenzyl ammonium chloride (TDMAB), cetylpyridinium chloride(CPCM) and dodecyl dimethylbenzyl ammonium chloride (DDAC) to produce TDMBA-I3, CPCM-I3 and DDAC-I3 association particles that exhibited the strongest resonance scattering (RS) peak at 460 nm, 540 nm and 430 nm, respectively. The G concentration in the range of 0.2~20×10-7 mol/L, 0.2~60×10-7 mol/L and 0.2~40×10-7 mol/L was linear to their RS intensity at the strongest RS wavelength, with a detection limit of 8.6×10-9 mol/L, 1.2×10-8 mol/L and 9.6×10-9 mol/L respectively. The TDMAB catalytic RS method was used to the determination of glucose in human serum, with satisfactory results.
Keywords/Search Tags:α-Amylase, Hydrogen Peroxide, Glucose oxidase, Glucose, Resonance scattering spectral
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