Analysis of 4-hydroxyproline using 4-chloro-7-nitrobenzo-2-oxa-1,3-diazol derivatization and micellar electrokinetic chromatography combined with laser-induced fluorescence detection.
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.
Dugan
M E
ME
Meat Research Section, Agriculture & Agri-Food Canada, Lacombe Research Centre. DuganM@em.agr.ca
Thacker
R D
RD
Aalhus
J L
JL
Jermiah
L E
LE
Lien
K A
KA
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D006868:Hydrolysis; D006909:Hydroxyproline / Q000032:analysis*; D007834:Lasers; D009578:Nitrobenzenes / Q000032:analysis*; D010069:Oxadiazoles / Q000032:analysis*; D012015:Reference Standards; D012995:Solubility; D013050:Spectrometry, Fluorescence / Q000379:methods
D000596:Amino Acids; D009578:Nitrobenzenes; D010069:Oxadiazoles; D006909:Hydroxyproline