Clinical Chemistry
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Clinical Chemistry 40: 1735-1738, 1994;
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Clinical Chemistry, Vol 40, 1735-1738, Copyright © 1994 by American Association for Clinical Chemistry

Improved determination of small amounts of free hydroxyproline in biological fluids

M Lange and M Malyusz
Physiologisches Institut der Universitat Kiel, Germany.

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.




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Copyright © 1994 by the American Association for Clinical Chemistry.