Journal
PIGMENT CELL & MELANOMA RESEARCH
Volume 24, Issue 4, Pages 605-613Publisher
WILEY
DOI: 10.1111/j.1755-148X.2011.00864.x
Keywords
eumelanin; pheomelanin; melanin; hair; oxidation; hydrogen peroxide
Categories
Funding
- Japan Society for the Promotion of Science (JSPS) [20591357, 21500358]
- Grants-in-Aid for Scientific Research [21500358] Funding Source: KAKEN
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Eumelanin and pheomelanin in tissue samples can be specifically measured as the markers pyrrole-2,3,5-tricarboxylic acid (PTCA) and 4-amino-3-hydroxyphenylalanine after acidic permanganate oxidation and hydroiodic acid hydrolysis, respectively. Those degradation methods, although widely applied, are not easily performed in most laboratories. To overcome this difficulty, we developed alkaline H2O2 oxidation in 1 M K2CO3 that produces, in addition to the eumelanin marker PTCA, thiazole-2,4,5-tricarboxylic acid (TTCA) and thiazole-4,5-dicarboxylic acid (TDCA) as markers for pheomelanin and pyrrole-2,3-dicarboxylic acid (PDCA) as a marker for 5,6-dihydroxyindole-derived eumelanin. Those four degradation products can be easily separated by HPLC and analyzed with ultraviolet detection. The alkaline H2O2 oxidation method is simple, reproducible and applicable to all pigmented tissues. Its application to characterize eumelanin and pheomelanin in human hair shows that PTCA and TTCA serve as specific markers for eumelanin and pheomelanin, respectively, although some caution is needed regarding the artificial production of TTCA from eumelanic tissue proteins.
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