Detection deoxyribose assay

Aruoma, O. 1. (1994). Deoxyribose assay for detecting hydroxyl radicals. Methods in Enzymology, Vol. 233, pp. 57-66, ISBN 978-0-12-182148-7. 

Aruoma Ol (1994) Deoxyribose assay for detecting hydroxyl radicals. Methods Enzymol 233 57-66 Ashton L, Buxton GV, Stuart CR (1995) Temperature dependence of the rate of reaction of OFI with some aromatic compounds in aqueous solution. J Chem Soc Faraday Trans 91 1631-1633 Asmus K-D, Mockel H, Flenglein A (1973) Pulse radiolytic study of the site of OFI radical attack on aliphatic alcohols in aqueous solution. J Phys Chem 77 1218-1221... 

Deoxyribose assay for the detection of hydroxyl radical production in cellular systems... 

An alternative method for free sialic acids only was introduced by Murayama et al (1976), in which reaction with pyridoxamine after heating at 70 °C for 45 min is followed by measurement of the fluorescence with 395 nm for excitation and 470 nm for emission. The method readily allows detection of 0.1 [xg of sialic acid and is not influenced by deoxyribose. However, a-keto acids develop the same fluorescence, and care must be taken to eliminate these compounds before using the assay (Murayama et al 1976). 

A modification of the periodic acid/thiobarbituric acid method has been described by excitation of the usual chromophore at 550 nm and measurement of the emission at 570 nm (Hammond and Papermaster 1976). The assay is 500-fold more sensitive than the conventional spectrophotometric procedures and can detect lOng sialic acid. Deoxyribose remains a problem with this assay and the precautions detailed for the periodic acid/thiobarbituric acid assay are required if accurate quantitation is to be made (Hammond and Papermaster 1976). 

An automated assay based on the Aminoff method, including acidic butan-l-ol and combining the sensitivity of the assay with ion-exchange purification, has been put forward by Krantz and Lee (1975) and remains the most sensitive of all automated methods. It can detect 1.5 nmoles of sialic acid ( 0.5 xg), eliminates major interference due to 2-deoxyribose and precludes any corrections for the presence of such interference. 

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