A study of the chemiluminescence behavior of myoglobin with luminol and its analytical applications

Zhenghua Song; Lin Wang; Shuang Hou
January 2004
Analytical & Bioanalytical Chemistry;Jan2004, Vol. 378 Issue 2, p529
Academic Journal
A chemiluminescence signal at 425 nm was observed when ferric state myoglobin was mixed with luminol in alkaline medium. Because the signal was remarkably enhanced in the presence of Fe(CN)64-, analytical applications were investigated in a flow-injection system. The increase in chemiluminescence was linearly dependent on myoglobin concentration in the range 0.1 to 100 nmol L-1, and the limit of detection was 0.04 nmol L-1 with relative standard deviation 3.2% (3σ). It was also found that binding of Mb with the ligands CN-, SCN-, and F- significantly inhibited the chemiluminescence reaction. The linear dynamic ranges for the ligands were 1.0–300.0, 0.1–3.0, and 0.5–100.0 nmol L-1, and the limits of detection (S/N=3) 0.4, 0.04, and 0.2 nmol L-1, for F-, CN-, and SCN-, respectively. The relative standard deviations were 5.32%, 6.13%, and 3.38% for 0.1 nmol L-1 CN-, 0.5 nmol L-1 SCN-, and 1.0 nmol L-1 F-, respectively. At a flow rate of 2.0 mL min-1 the assay could be accomplished in 1 min, including sampling and washing. The method has been successfully applied to the determination of myoglobin in human urine and F- in water samples. A possible mechanism of chemiluminescence production by myoglobin and luminol is presented.


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