Assessment of acyl groups and reaction conditions in the competition between perhydrolysis and hydrolysis of acyl resorufins for developing an indicator reaction for fluorometric analysis of hydrogen peroxide
Chemical and Pharmaceutical Bulletin, ISSN: 0009-2363, Vol: 50, Issue: 2, Page: 169-174
2002
- 8Citations
- 12Captures
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Article Description
Perhydrolysis of acetyl resorufin (AR) was reported previously to work as a fluorometric indicator reaction for glucose determination using only glucose oxidase. However, hydrolysis of AR in blank solution rendered the working concentration range of this method less than two orders of magnitude. To exclude or at least significantly reduce this interference, acyl groups and reaction conditions in the competition between perhydrolysis and hydrolysis of various acyl resorufins were assessed. Fluorometric evaluation of reactions in the presence or absence of HO in phosphate buffer (pH 7.5, 100 mM)-CHCN at 25°C demonstrated that in tert-butylacetyl, isobutyryl, cyclohexanecarbonyl and pivaloyl resorufins (TBAR, IBR, CHR and PVR, respectively) among 10 acyl resorufins examined here, the competitive situation was shifted in a much more favorable way to perhydrolysis than in AR, although fluorometric responses due to their HO-dependent deacylation were suppressed in comparison with AR. Examination of the effects of pH, components and concentrations of buffers as well as reaction temperature established reaction conditions that not only allowed perhydrolysis of each of these four compounds to prevail over hydrolysis more effectively, but also improved the HO-based fluorometric responses. Thus, perhydrolysis of TBAR, IBR, CHR and PVR in phosphate buffer (pH 8.0, 20 mM)-CHCN at 25°C worked effectively as fluorometric indicator reactions for HO analysis, affording a calibration curve over a concentration range of three orders of magnitude. Taking sensitivity, reproducibility and the response for blank solution into consideration, PVR seemed to be the best choice as a fluorochromogen for HO determination under these conditions. For HO analysis at lower pH, perhydrolysis of IBR in phosphate buffer (pH 7.5, 20 mM)-CH CN was shown to effectively function as an indicator reaction. Applicability of the fluorometric methods with PVR and IBR to blood glucose determination was also discussed, comparing with Trinder's method with phenol, 4-aminoantipyrine and peroxidase (POD). © 2002 Pharmaceutical Society of Japan.
Bibliographic Details
http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0036483630&origin=inward; http://dx.doi.org/10.1248/cpb.50.169; http://www.ncbi.nlm.nih.gov/pubmed/11848204; http://www.jstage.jst.go.jp/article/cpb/50/2/50_2_169/_article; https://dx.doi.org/10.1248/cpb.50.169; https://www.jstage.jst.go.jp/article/cpb/50/2/50_2_169/_article
Pharmaceutical Society of Japan
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