In the presence of ferrous ions (Fe2+), the anti-tumour agent bleomycin will induce DNA degradation. Degradation of DNA into substances detectable by the thiobarbituric acid test has been used previously for the detection of iron in a form that is capable of catalysing the formation of the potentially harmful hydroxyl free radical. In the present paper, we describe the application of the ethidium-binding assay of DNA damage to the measurement of bleomycin-detectable iron, comparing its performance with the conventional method in the assessment of iron standard solutions and plasma samples from haemochromatosis patients. The ethidium-binding assay proved to be more responsive than the thiobarbituric acid test in the detection of DNA damage induced by very low concentrations of iron, but became saturated at higher iron concentrations. Agreement between the two versions of the assay in the identification of plasma samples containing bleomycin-detectable iron was good, but agreement on the actual concentrations of such iron in the positive samples was poor. This discrepancy is believed to be due to interference with the thiobarbituric acid assay by plasma. Consequently, it was not possible to obtain reliable estimates of free iron concentrations in plasma when using the conventional version of the bleomycin assay. We have devised a parameter of iron status called the catalytic iron index. For healthy, non-haemochromatotic individuals, the mean value of this parameter was found to be 0.81 (range 0.78–0.84; n = 20). Elevated values were observed in some plasma samples from haemochromatosis patients, but these showed no correlation with serum ferritin levels. In contrast, correlations were seen with both serum iron and transferrin saturation levels, but only when these were above the normal range.
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January 24 2001
A simple, highly sensitive and improved method for the measurement of bleomycin-detectable iron: the ‘catalytic iron index’ and its value in the assessment of iron status in haemochromatosis
Mark J. BURKITT;
*Gray Laboratory Cancer Research Trust, P.O. Box 100, Mount Vernon Hospital, Northwood, Middlesex HA6 2JR, U.K.
Correspondence: Dr M. J. Burkitt (e-mail [email protected])
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Lesley MILNE;
Lesley MILNE
†Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, Scotland, U.K.
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Alaeddin RAAFAT
Alaeddin RAAFAT
‡Department of Haematology, Hairmyres Hospital, East Kilbride G75 8RJ, Scotland, U.K.
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Publisher: Portland Press Ltd
Received:
August 23 2000
Revision Received:
September 22 2000
Accepted:
November 09 2000
Online ISSN: 1470-8736
Print ISSN: 0143-5221
The Biochemical Society and the Medical Research Society © 2001
2001
Clin Sci (Lond) (2001) 100 (3): 239–247.
Article history
Received:
August 23 2000
Revision Received:
September 22 2000
Accepted:
November 09 2000
Citation
Mark J. BURKITT, Lesley MILNE, Alaeddin RAAFAT; A simple, highly sensitive and improved method for the measurement of bleomycin-detectable iron: the ‘catalytic iron index’ and its value in the assessment of iron status in haemochromatosis. Clin Sci (Lond) 1 March 2001; 100 (3): 239–247. doi: https://doi.org/10.1042/cs1000239
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