Improved determination of small amounts of free hydroxyproline in biological fluids.
A fast, reliable, and sensitive (< 20 pmol) method for the quantification of 4-hydroxyproline is described. It ensures good separation of imino acid peaks, eliminates interference by primary amino acids, and guarantees full (96-105%) recovery of hydroxyproline (HYP). Interfering primary amino acids are derivatized by o-phthaldialdehyde and removed from the sample by use of a discardable C18 column. HYP is measured photometrically at 254 nm after a second derivatization with phenylisothiocyanate and isocratic separation on a reversed-phase HPLC column.
Lange
M
M
Physiologisches Institut der Universität Kiel, Germany.
Mályusz
M
M
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D006909:Hydroxyproline / Q000032:analysis* / Q000097:blood / Q000652:urine; D017879:Isothiocyanates; D013048:Specimen Handling; D013056:Spectrophotometry, Ultraviolet; D013861:Thiocyanates
D017879:Isothiocyanates; D013861:Thiocyanates; C005441:phenylisothiocyanate; D006909:Hydroxyproline