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Analysis of 4-hydroxyproline using 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol derivatization and micellar electrokinetic
M E Dugan1, R D Thacker, J L Aalhus
1Meat Research Section, Agriculture & Agri-Food Canada, Lacombe Research Centre. DuganM@em.agr.ca
Journal of Chromatography. B, Biomedical Sciences and Applications
|September 14, 2000
Abstract:
Micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence (LIF) detection was used for internal standard (pyrrolidinol) based quantification of 4-hydroxyproline (Hyp) in muscle hydrolysates. Hydrolysates were first derivatized with o-phthaldialdehyde to reduce primary amine interference and then 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol (NBD) to enable selective detection of secondary amines. This method allows for rapid and sensitive detection of hydroxyproline in dilute hydrolysates using commercially available capillary electrophoresis equipment.